Baxter, J.S.
Brough, R.
Krastev, D.B.
Song, F.
Sridhar, S.
Gulati, A.
Alexander, J.
Roumeliotis, T.I.
Kozik, Z.
Choudhary, J.S.
Haider, S.
Pettitt, S.J.
Tutt, A.N.
Lord, C.J.
(2024). Cancer-associated FBXW7 loss is synthetic lethal with pharmacological targeting of CDC7. Mol oncol,
Vol.18
(2),
pp. 369-385.
show abstract
full text
The F-box and WD repeat domain containing 7 (FBXW7) tumour suppressor gene encodes a substrate-recognition subunit of Skp, cullin, F-box (SCF)-containing complexes. The tumour-suppressive role of FBXW7 is ascribed to its ability to drive ubiquitination and degradation of oncoproteins. Despite this molecular understanding, therapeutic approaches that target defective FBXW7 have not been identified. Using genome-wide clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 screens, focussed RNA-interference screens and whole and phospho-proteome mass spectrometry profiling in multiple FBXW7 wild-type and defective isogenic cell lines, we identified a number of FBXW7 synthetic lethal targets, including proteins involved in the response to replication fork stress and proteins involved in replication origin firing, such as cell division cycle 7-related protein kinase (CDC7) and its substrate, DNA replication complex GINS protein SLD5 (GINS4). The CDC7 synthetic lethal effect was confirmed using small-molecule inhibitors. Mechanistically, FBXW7/CDC7 synthetic lethality is dependent upon the replication factor telomere-associated protein RIF1 (RIF1), with RIF1 silencing reversing the FBXW7-selective effects of CDC7 inhibition. The delineation of FBXW7 synthetic lethal effects we describe here could serve as the starting point for subsequent drug discovery and/or development in this area..
Loibl, S.
André, F.
Bachelot, T.
Barrios, C.H.
Bergh, J.
Burstein, H.J.
Cardoso, M.J.
Carey, L.A.
Dawood, S.
Del Mastro, L.
Denkert, C.
Fallenberg, E.M.
Francis, P.A.
Gamal-Eldin, H.
Gelmon, K.
Geyer, C.E.
Gnant, M.
Guarneri, V.
Gupta, S.
Kim, S.B.
Krug, D.
Martin, M.
Meattini, I.
Morrow, M.
Janni, W.
Paluch-Shimon, S.
Partridge, A.
Poortmans, P.
Pusztai, L.
Regan, M.M.
Sparano, J.
Spanic, T.
Swain, S.
Tjulandin, S.
Toi, M.
Trapani, D.
Tutt, A.
Xu, B.
Curigliano, G.
Harbeck, N.
ESMO Guidelines Committee. Electronic address: [email protected],
(2024). Early breast cancer: ESMO Clinical Practice Guideline for diagnosis, treatment and follow-up. Ann oncol,
Vol.35
(2),
pp. 159-182.
Harvey-Jones, E.
Raghunandan, M.
Robbez-Masson, L.
Magraner-Pardo, L.
Alaguthurai, T.
Yablonovitch, A.
Yen, J.
Xiao, H.
Brough, R.
Frankum, J.
Song, F.
Yeung, J.
Savy, T.
Gulati, A.
Alexander, J.
Kemp, H.
Starling, C.
Konde, A.
Marlow, R.
Cheang, M.
Proszek, P.
Hubank, M.
Cai, M.
Trendell, J.
Lu, R.
Liccardo, R.
Ravindran, N.
Llop-Guevara, A.
Rodriguez, O.
Balmana, J.
Lukashchuk, N.
Dorschner, M.
Drusbosky, L.
Roxanis, I.
Serra, V.
Haider, S.
Pettitt, S.J.
Lord, C.J.
Tutt, A.N.
(2024). Longitudinal profiling identifies co-occurring BRCA1/2 reversions, TP53BP1, RIF1 and PAXIP1 mutations in PARP inhibitor-resistant advanced breast cancer. Ann oncol,
Vol.35
(4),
pp. 364-380.
show abstract
full text
BACKGROUND: Resistance to therapies that target homologous recombination deficiency (HRD) in breast cancer limits their overall effectiveness. Multiple, preclinically validated, mechanisms of resistance have been proposed, but their existence and relative frequency in clinical disease are unclear, as is how to target resistance. PATIENTS AND METHODS: Longitudinal mutation and methylation profiling of circulating tumour (ct)DNA was carried out in 47 patients with metastatic BRCA1-, BRCA2- or PALB2-mutant breast cancer treated with HRD-targeted therapy who developed progressive disease-18 patients had primary resistance and 29 exhibited response followed by resistance. ctDNA isolated at multiple time points in the patient treatment course (before, on-treatment and at progression) was sequenced using a novel >750-gene intron/exon targeted sequencing panel. Where available, matched tumour biopsies were whole exome and RNA sequenced and also used to assess nuclear RAD51. RESULTS: BRCA1/2 reversion mutations were present in 60% of patients and were the most prevalent form of resistance. In 10 cases, reversions were detected in ctDNA before clinical progression. Two new reversion-based mechanisms were identified: (i) intragenic BRCA1/2 deletions with intronic breakpoints; and (ii) intragenic BRCA1/2 secondary mutations that formed novel splice acceptor sites, the latter being confirmed by in vitro minigene reporter assays. When seen before commencing subsequent treatment, reversions were associated with significantly shorter time to progression. Tumours with reversions retained HRD mutational signatures but had functional homologous recombination based on RAD51 status. Although less frequent than reversions, nonreversion mechanisms [loss-of-function (LoF) mutations in TP53BP1, RIF1 or PAXIP1] were evident in patients with acquired resistance and occasionally coexisted with reversions, challenging the notion that singular resistance mechanisms emerge in each patient. CONCLUSIONS: These observations map the prevalence of candidate drivers of resistance across time in a clinical setting, information with implications for clinical management and trial design in HRD breast cancers..
Ganz, P.A.
Bandos, H.
Španić, T.
Friedman, S.
Müller, V.
Kuemmel, S.
Delaloge, S.
Brain, E.
Toi, M.
Yamauchi, H.
de Dueñas, E.-.
Armstrong, A.
Im, S.-.
Song, C.-.
Zheng, H.
Sarosiek, T.
Sharma, P.
Geng, C.
Fu, P.
Rhiem, K.
Frauchiger-Heuer, H.
Wimberger, P.
t'Kint de Roodenbeke, D.
Liao, N.
Goodwin, A.
Chakiba-Brugère, C.
Friedlander, M.
Lee, K.S.
Giacchetti, S.
Takano, T.
Henao-Carrasco, F.
Virani, S.
Valdes-Albini, F.
Domchek, S.M.
Bane, C.
McCarron, E.C.
Mita, M.
Rossi, G.
Rastogi, P.
Fielding, A.
Gelber, R.D.
Scheepers, E.D.
Cameron, D.
Garber, J.
Geyer, C.E.
Tutt, A.N.
(2024). Patient-Reported Outcomes in OlympiA: A Phase III, Randomized, Placebo-Controlled Trial of Adjuvant Olaparib in gBRCA1/2 Mutations and High-Risk Human Epidermal Growth Factor Receptor 2-Negative Early Breast Cancer. J clin oncol,
Vol.42
(11),
pp. 1288-1300.
show abstract
PURPOSE: The OlympiA randomized phase III trial compared 1 year of olaparib (OL) or placebo (PL) as adjuvant therapy in patients with germline BRCA1/2, high-risk human epidermal growth factor receptor 2-negative early breast cancer after completing (neo)adjuvant chemotherapy ([N]ACT), surgery, and radiotherapy. The patient-reported outcome primary hypothesis was that OL-treated patients may experience greater fatigue during treatment. METHODS: Data were collected before random assignment, and at 6, 12, 18, and 24 months. The primary end point was fatigue, measured with the Functional Assessment of Chronic Illness Therapy-Fatigue scale. Secondary end points, assessed with the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire, Core 30 item, included nausea and vomiting (NV), diarrhea, and multiple functional domains. Scores were compared between treatment groups using mixed model for repeated measures. Two-sided P values <.05 were statistically significant for the primary end point. All secondary end points were descriptive. RESULTS: One thousand five hundred and thirty-eight patients (NACT: 746, ACT: 792) contributed to the analysis. Fatigue severity was statistically significantly greater for OL versus PL, but not clinically meaningfully different by prespecified criteria (≥3 points) at 6 months (diff OL v PL: NACT: -1.3 [95% CI, -2.4 to -0.2]; P = .022; ACT: -1.3 [95% CI, -2.3 to -0.2]; P = .017) and 12 months (NACT: -1.6 [95% CI, -2.8 to -0.3]; P = .017; ACT: -1.3 [95% CI, -2.4 to -0.2]; P = .025). There were no significant differences in fatigue severity between treatment groups at 18 and 24 months. NV severity was worse in patients treated with OL compared with PL at 6 months (NACT: 6.0 [95% CI, 4.1 to 8.0]; ACT: 5.3 [95% CI, 3.4 to 7.2]) and 12 months (NACT: 6.4 [95% CI, 4.4 to 8.3]; ACT: 4.5 [95% CI, 2.8 to 6.1]). During treatment, there were some clinically meaningful differences between groups for other symptoms but not for function subscales or global health status. CONCLUSION: Treatment-emergent symptoms from OL were limited, generally resolving after treatment ended. OL- and PL-treated patients had similar functional scores, slowly improving during the 24 months after (N)ACT and there was no clinically meaningful persistence of fatigue severity in OL-treated patients..
Harvey-Jones, E.J.
Lord, C.J.
Tutt, A.N.
(2023). Systemic Therapy for Hereditary Breast Cancers. Hematol oncol clin north am,
Vol.37
(1),
pp. 203-224.
show abstract
full text
Approximately 5% to 10% of all breast cancers are hereditary; many of which are caused by pathogenic variants in genes required for homologous recombination, including BRCA1 and BRCA2. Here we discuss systemic treatment for such breast cancers, including approved chemotherapeutic approaches and also targeted treatment approaches using poly-(ADP ribose) polymerase inhibitors. We also discuss experimental approaches to treating hereditary breast cancer, including new small molecule DNA repair inhibitors and also immunomodulatory agents. Finally, we discuss how drug resistance emerges in patients with hereditary breast cancer, how this might be delayed or prevented, and how biomarker-adapted treatment is molding the future management of hereditary breast cancer..
Turner, N.C.
Swift, C.
Jenkins, B.
Kilburn, L.
Coakley, M.
Beaney, M.
Fox, L.
Goddard, K.
Garcia-Murillas, I.
Proszek, P.
Hall, P.
Harper-Wynne, C.
Hickish, T.
Kernaghan, S.
Macpherson, I.R.
Okines, A.F.
Palmieri, C.
Perry, S.
Randle, K.
Snowdon, C.
Stobart, H.
Wardley, A.M.
Wheatley, D.
Waters, S.
Winter, M.C.
Hubank, M.
Allen, S.D.
Bliss, J.M.
c-TRAK TN investigators,
(2023). Results of the c-TRAK TN trial: a clinical trial utilising ctDNA mutation tracking to detect molecular residual disease and trigger intervention in patients with moderate- and high-risk early-stage triple-negative breast cancer. Ann oncol,
Vol.34
(2),
pp. 200-211.
show abstract
full text
BACKGROUND: Post-treatment detection of circulating tumour DNA (ctDNA) in early-stage triple-negative breast cancer (TNBC) patients predicts high risk of relapse. c-TRAK TN assessed the utility of prospective ctDNA surveillance in TNBC and the activity of pembrolizumab in patients with ctDNA detected [ctDNA positive (ctDNA+)]. PATIENTS AND METHODS: c-TRAK TN, a multicentre phase II trial, with integrated prospective ctDNA surveillance by digital PCR, enrolled patients with early-stage TNBC and residual disease following neoadjuvant chemotherapy, or stage II/III with adjuvant chemotherapy. ctDNA surveillance comprised three-monthly blood sampling to 12 months (18 months if samples were missed due to coronavirus disease), and ctDNA+ patients were randomised 2 : 1 to intervention : observation. ctDNA results were blinded unless patients were allocated to intervention, when staging scans were done and those free of recurrence were offered pembrolizumab. A protocol amendment (16 September 2020) closed the observation group; all subsequent ctDNA+ patients were allocated to intervention. Co-primary endpoints were (i) ctDNA detection rate and (ii) sustained ctDNA clearance rate on pembrolizumab (NCT03145961). RESULTS: Two hundred and eight patients registered between 30 January 2018 and 06 December 2019, 185 had tumour sequenced, 171 (92.4%) had trackable mutations, and 161 entered ctDNA surveillance. Rate of ctDNA detection by 12 months was 27.3% (44/161, 95% confidence interval 20.6% to 34.9%). Seven patients relapsed without prior ctDNA detection. Forty-five patients entered the therapeutic component (intervention n = 31; observation n = 14; one observation patient was re-allocated to intervention following protocol amendment). Of patients allocated to intervention, 72% (23/32) had metastases on staging at the time of ctDNA+, and 4 patients declined pembrolizumab. Of the five patients who commenced pembrolizumab, none achieved sustained ctDNA clearance. CONCLUSIONS: c-TRAK TN is the first prospective study to assess whether ctDNA assays have clinical utility in guiding therapy in TNBC. Patients had a high rate of metastatic disease on ctDNA detection. Findings have implications for future trial design, emphasising the importance of commencing ctDNA testing early, with more sensitive and/or frequent ctDNA testing regimes..
Yamauchi, H.
Toi, M.
Takayama, S.
Nakamura, S.
Takano, T.
Cui, K.
Campbell, C.
De Vos, L.
Geyer, C.
Tutt, A.
(2023). Adjuvant olaparib in the subset of patients from Japan with BRCA1- or BRCA2-mutated high-risk early breast cancer from the phase 3 OlympiA trial. Breast cancer,
Vol.30
(4),
pp. 596-605.
show abstract
full text
BACKGROUND: The efficacy and safety of olaparib compared with placebo in the subset of patients from Japan in the phase 3 OlympiA trial (NCT02032823) are reported here and contextualized with reference to the global OlympiA population. METHODS: Patients with germline BRCA1 and/or BRCA2 pathogenic variants and HER2-negative, high-risk early breast cancer who had received neoadjuvant or adjuvant chemotherapy and completed local treatment were eligible. Patients were randomized 1:1 to receive olaparib or placebo for 1 year. PRIMARY ENDPOINT: invasive disease-free survival (IDFS). Secondary endpoints: distant disease-free survival (DDFS), overall survival (OS), and safety. Data are reported from the first pre-specified interim analysis (data cut-off [DCO] March 27, 2020) and the second, event driven, pre-specified interim analysis of OS (DCO July 12, 2021) in patients from Japan. RESULTS: 140 patients were randomized in Japan (olaparib, n = 64; placebo, n = 76). At the first pre-specified interim analysis (median follow-up: 2.9 years), hazard ratios (HRs) for adjuvant olaparib compared with placebo were 0.5 for IDFS (95% confidence interval [CI] 0.18-1.24) and 0.41 for DDFS (95% CI 0.11-1.16). At the second pre-specified interim analysis of OS, three deaths occurred in the olaparib group versus six deaths in the placebo group (HR, 0.62 [95% CI 0.13-2.36]). Findings were consistent with those for the global population. No new safety signals were observed. CONCLUSIONS: While this analysis in a Japanese subset of patients was not powered to detect population-related treatment differences, efficacy and safety analysis results were consistent with the global OlympiA population, suggesting the findings from the global study are generalizable to clinical practice in Japan..
Yeung, J.
Fotiadis, N.
Diamantopoulos, A.
Tutt, A.
Roxanis, I.
Bandula, S.
(2023). Next-Generation Sequencing and Image-Guided Tissue Sampling: A Primer for Interventional Radiologists. J vasc interv radiol,
Vol.34
(8),
pp. 1291-1302.e1.
show abstract
full text
The discovery of increasing numbers of actionable molecular and gene targets for cancer treatment has driven the demand for tissue sampling for next-generation sequencing (NGS). Requirements for sequencing can be very specific, and inadequate sampling leads to delays in management and decision making. It is important that interventional radiologists are aware of NGS technologies and their common applications and be cognizant of the factors that contribute to successful sample sequencing. This review summarizes the fundamentals of cancer tissue collection and processing for NGS. It elaborates on sequencing technologies and their applications with the aim of providing readers with a working understanding that can enhance their clinical practice. It then describes imaging, tumor, biopsy, and sample collection factors that improve the chances of NGS success. Finally, it discusses future practice, highlighting the problem of undersampling in both clinical and research settings and the opportunities within interventional radiology to address this..
Pettitt, S.J.
Shao, N.
Zatreanu, D.
Frankum, J.
Bajrami, I.
Brough, R.
Krastev, D.B.
Roumeliotis, T.I.
Choudhary, J.S.
Lorenz, S.
Rust, A.
de Bono, J.S.
Yap, T.A.
Tutt, A.N.
Lord, C.J.
(2023). A HUWE1 defect causes PARP inhibitor resistance by modulating the BRCA1-∆11q splice variant. Oncogene,
Vol.42
(36),
pp. 2701-2709.
show abstract
full text
Although PARP inhibitors (PARPi) now form part of the standard-of-care for the treatment of homologous recombination defective cancers, de novo and acquired resistance limits their overall effectiveness. Previously, overexpression of the BRCA1-∆11q splice variant has been shown to cause PARPi resistance. How cancer cells achieve increased BRCA1-∆11q expression has remained unclear. Using isogenic cells with different BRCA1 mutations, we show that reduction in HUWE1 leads to increased levels of BRCA1-∆11q and PARPi resistance. This effect is specific to cells able to express BRCA1-∆11q (e.g. BRCA1 exon 11 mutant cells) and is not seen in BRCA1 mutants that cannot express BRCA1-∆11q, nor in BRCA2 mutant cells. As well as increasing levels of BRCA1-∆11q protein in exon 11 mutant cells, HUWE1 silencing also restores RAD51 nuclear foci and platinum salt resistance. HUWE1 catalytic domain mutations were also seen in a case of PARPi resistant, BRCA1 exon 11 mutant, high grade serous ovarian cancer. These results suggest how elevated levels of BRCA1-∆11q and PARPi resistance can be achieved, identify HUWE1 as a candidate biomarker of PARPi resistance for assessment in future clinical trials and illustrate how some PARPi resistance mechanisms may only operate in patients with particular BRCA1 mutations..
Tovey, H.
Sipos, O.
Parker, J.S.
Hoadley, K.A.
Quist, J.
Kernaghan, S.
Kilburn, L.
Salgado, R.
Loi, S.
Kennedy, R.D.
Roxanis, I.
Gazinska, P.
Pinder, S.E.
Bliss, J.
Perou, C.M.
Haider, S.
Grigoriadis, A.
Tutt, A.
Cheang, M.C.
(2023). Integrated Multimodal Analyses of DNA Damage Response and Immune Markers as Predictors of Response in Metastatic Triple-Negative Breast Cancer in the TNT Trial (NCT00532727). Clin cancer res,
Vol.29
(18),
pp. 3691-3705.
show abstract
full text
PURPOSE: The TNT trial (NCT00532727) showed no evidence of carboplatin superiority over docetaxel in metastatic triple-negative breast cancer (mTNBC), but carboplatin benefit was observed in the germline BRCA1/2 mutation subgroup. Broader response-predictive biomarkers are needed. We explored the predictive ability of DNA damage response (DDR) and immune markers. EXPERIMENTAL DESIGN: Tumor-infiltrating lymphocytes were evaluated for 222 of 376 patients. Primary tumors (PT) from 186 TNT participants (13 matched recurrences) were profiled using total RNA sequencing. Four transcriptional DDR-related and 25 immune-related signatures were evaluated. We assessed their association with objective response rate (ORR) and progression-free survival (PFS). Conditional inference forest clustering was applied to integrate multimodal data. The biology of subgroups was characterized by 693 gene expression modules and other markers. RESULTS: Transcriptional DDR-related biomarkers were not predictive of ORR to either treatment overall. Changes from PT to recurrence were demonstrated; in chemotherapy-naïve patients, transcriptional DDR markers separated carboplatin responders from nonresponders (P values = 0.017; 0.046). High immune infiltration was associated with docetaxel ORR (interaction P values < 0.05). Six subgroups were identified; the immune-enriched cluster had preferential docetaxel response [62.5% (D) vs. 29.4% (C); P = 0.016]. The immune-depleted cluster had preferential carboplatin response [8.0% (D) vs. 40.0% (C); P = 0.011]. DDR-related subgroups were too small to assess ORR. CONCLUSIONS: High immune features predict docetaxel response, and high DDR signature scores predict carboplatin response in treatment-naïve mTNBC. Integrating multimodal DDR and immune-related markers identifies subgroups with differential treatment sensitivity. Treatment options for patients with immune-low and DDR-proficient tumors remains an outstanding need. Caution is needed using PT-derived transcriptional signatures to direct treatment in mTNBC, particularly DDR-related markers following prior chemotherapy..
Fitzpatrick, A.
Iravani, M.
Mills, A.
Vicente, D.
Alaguthurai, T.
Roxanis, I.
Turner, N.C.
Haider, S.
Tutt, A.N.
Isacke, C.M.
(2023). Genomic profiling and pre-clinical modelling of breast cancer leptomeningeal metastasis reveals acquisition of a lobular-like phenotype. Nat commun,
Vol.14
(1),
p. 7408.
show abstract
full text
Breast cancer leptomeningeal metastasis (BCLM), where tumour cells grow along the lining of the brain and spinal cord, is a devastating development for patients. Investigating this metastatic site is hampered by difficulty in accessing tumour material. Here, we utilise cerebrospinal fluid (CSF) cell-free DNA (cfDNA) and CSF disseminated tumour cells (DTCs) to explore the clonal evolution of BCLM and heterogeneity between leptomeningeal and extracranial metastatic sites. Somatic alterations with potential therapeutic actionability were detected in 81% (17/21) of BCLM cases, with 19% detectable in CSF cfDNA only. BCLM was enriched in genomic aberrations in adherens junction and cytoskeletal genes, revealing a lobular-like breast cancer phenotype. CSF DTCs were cultured in 3D to establish BCLM patient-derived organoids, and used for the successful generation of BCLM in vivo models. These data reveal that BCLM possess a unique genomic aberration profile and highlight potential cellular dependencies in this hard-to-treat form of metastatic disease..
Zelceski, A.
Francica, P.
Lingg, L.
Mutlu, M.
Stok, C.
Liptay, M.
Alexander, J.
Baxter, J.S.
Brough, R.
Gulati, A.
Haider, S.
Raghunandan, M.
Song, F.
Sridhar, S.
Forment, J.V.
O'Connor, M.J.
Davies, B.R.
van Vugt, M.A.
Krastev, D.B.
Pettitt, S.J.
Tutt, A.N.
Rottenberg, S.
Lord, C.J.
(2023). MND1 and PSMC3IP control PARP inhibitor sensitivity in mitotic cells. Cell rep,
Vol.42
(5),
p. 112484.
show abstract
full text
The PSMC3IP-MND1 heterodimer promotes meiotic D loop formation before DNA strand exchange. In genome-scale CRISPR-Cas9 mutagenesis and interference screens in mitotic cells, depletion of PSMC3IP or MND1 causes sensitivity to poly (ADP-Ribose) polymerase inhibitors (PARPi) used in cancer treatment. PSMC3IP or MND1 depletion also causes ionizing radiation sensitivity. These effects are independent of PSMC3IP/MND1's role in mitotic alternative lengthening of telomeres. PSMC3IP- or MND1-depleted cells accumulate toxic RAD51 foci in response to DNA damage, show impaired homology-directed DNA repair, and become PARPi sensitive, even in cells lacking both BRCA1 and TP53BP1. Epistasis between PSMC3IP-MND1 and BRCA1/BRCA2 defects suggest that abrogated D loop formation is the cause of PARPi sensitivity. Wild-type PSMC3IP reverses PARPi sensitivity, whereas a PSMC3IP p.Glu201del mutant associated with D loop defects and ovarian dysgenesis does not. These observations suggest that meiotic proteins such as MND1 and PSMC3IP have a greater role in mitotic DNA repair..
Graham, R.
Gazinska, P.
Zhang, B.
Khiabany, A.
Sinha, S.
Alaguthurai, T.
Flores-Borja, F.
Vicencio, J.
Beuron, F.
Roxanis, I.
Matkowski, R.
Liam-Or, R.
Tutt, A.
Ng, T.
Al-Jamal, K.T.
Zhou, Y.
Irshad, S.
(2023). Serum-derived extracellular vesicles from breast cancer patients contribute to differential regulation of T-cell-mediated immune-escape mechanisms in breast cancer subtypes. Front immunol,
Vol.14,
p. 1204224.
show abstract
full text
BACKGROUND: Intracellular communication within the tumour is complex and extracellular vesicles (EVs) have been identified as major contributing factors for the cell-to-cell communication in the local and distant tumour environments. Here, we examine the differential effects of breast cancer (BC) subtype-specific patient serum and cell-line derived EVs in the regulation of T cell mediated immune responses. METHODS: Ultracentrifugation was used to isolate EVs from sera of 63 BC patients, 15 healthy volunteers and 4 human breast cancer cell lines. Longitudinal blood draws for EV isolation for patients on neoadjuvant chemotherapy was also performed. Characterization of EVs was performed by Nanoparticle Tracking Analysis (NTA), transmission electron microscopy (TEM) and immunoblotting. CD63 staining was performed on a tissue microarray of 218 BC patients. In-house bioinformatics algorithms were utilized for the computation of EV associated expression scores within The Cancer Genome Atlas (TCGA) and correlated with tumour infiltrating lymphocyte (TIL) scores. In vitro stimulation of PBMCs with EVs from serum and cell-line derived EVs was performed and changes in the immune phenotypes characterized by flow cytometry. Cytokine profiles were assessed using a 105-plex immunoassay or IL10 ELISA. RESULTS: Patients with triple negative breast cancers (TNBCs) exhibited the lowest number of EVs in the sera; whilst the highest was detected in ER+HER2+ cancers; reflected also in the higher level of CD63+ vesicles found within the ER+HER2+ local tumour microenvironment. Transcriptomic analysis of the TCGA data identified that samples assigned with lower EV scores had significantly higher abundance of CD4+ memory activated T cells, T follicular cells and CD8 T cells, plasma, and memory B cells; whilst samples with high EV scores were more enriched for anti-inflammatory M2 macrophages and mast cells. A negative correlation between EV expression scores and stromal TIL counts was also observed. In vitro experiments confirmed that circulating EVs within breast cancer subtypes have functionally differing immunomodulatory capabilities, with EVs from patients with the most aggressive breast cancer subtype (TNBCs) demonstrating the most immune-suppressive phenotype (decreased CD3+HLA-DR+ but increased CD3+PD-L1 T cells, increased CD4+CD127-CD25hi T regulatory cells with associated increase in IL10 cytokine production). In depth assessment of the cytokine modulation triggered by the serum/cell line derived exosomes confirmed differential inflammatory cytokine profiles across differing breast cancer subtypes. Studies using the MDA-231 TNBC breast cancer cell-line derived EVs provided further support that TNBC EVs induced the most immunosuppressive response within PBMCs. DISCUSSION: Our study supports further investigations into how tumour derived EVs are a mechanism that cancers can exploit to promote immune suppression; and breast cancer subtypes produce EVs with differing immunomodulatory capabilities. Understanding the intracellular/extracellular pathways implicated in alteration from active to suppressed immune state may provide a promising way forward for restoring immune competence in specific breast cancer patient populations..
Krastev, D.B.
Li, S.
Sun, Y.
Wicks, A.J.
Hoslett, G.
Weekes, D.
Badder, L.M.
Knight, E.G.
Marlow, R.
Pardo, M.C.
Yu, L.
Talele, T.T.
Bartek, J.
Choudhary, J.S.
Pommier, Y.
Pettitt, S.J.
Tutt, A.N.
Ramadan, K.
Lord, C.J.
(2022). The ubiquitin-dependent ATPase p97 removes cytotoxic trapped PARP1 from chromatin. Nat cell biol,
Vol.24
(1),
pp. 62-73.
show abstract
full text
Poly (ADP-ribose) polymerase (PARP) inhibitors elicit antitumour activity in homologous recombination-defective cancers by trapping PARP1 in a chromatin-bound state. How cells process trapped PARP1 remains unclear. Using wild-type and a trapping-deficient PARP1 mutant combined with rapid immunoprecipitation mass spectrometry of endogenous proteins and Apex2 proximity labelling, we delineated mass spectrometry-based interactomes of trapped and non-trapped PARP1. These analyses identified an interaction between trapped PARP1 and the ubiquitin-regulated p97 ATPase/segregase. We found that following trapping, PARP1 is SUMOylated by PIAS4 and subsequently ubiquitylated by the SUMO-targeted E3 ubiquitin ligase RNF4, events that promote recruitment of p97 and removal of trapped PARP1 from chromatin. Small-molecule p97-complex inhibitors, including a metabolite of the clinically used drug disulfiram (CuET), prolonged PARP1 trapping and enhanced PARP inhibitor-induced cytotoxicity in homologous recombination-defective tumour cells and patient-derived tumour organoids. Together, these results suggest that p97 ATPase plays a key role in the processing of trapped PARP1 and the response of tumour cells to PARP inhibitors..
Rätze, M.A.
Koorman, T.
Sijnesael, T.
Bassey-Archibong, B.
van de Ven, R.
Enserink, L.
Visser, D.
Jaksani, S.
Viciano, I.
Bakker, E.R.
Richard, F.
Tutt, A.
O'Leary, L.
Fitzpatrick, A.
Roca-Cusachs, P.
van Diest, P.J.
Desmedt, C.
Daniel, J.M.
Isacke, C.M.
Derksen, P.W.
(2022). Loss of E-cadherin leads to Id2-dependent inhibition of cell cycle progression in metastatic lobular breast cancer. Oncogene,
Vol.41
(21),
pp. 2932-2944.
show abstract
full text
Invasive lobular breast carcinoma (ILC) is characterized by proliferative indolence and long-term latency relapses. This study aimed to identify how disseminating ILC cells control the balance between quiescence and cell cycle re-entry. In the absence of anchorage, ILC cells undergo a sustained cell cycle arrest in G0/G1 while maintaining viability. From the genes that are upregulated in anchorage independent ILC cells, we selected Inhibitor of DNA binding 2 (Id2), a mediator of cell cycle progression. Using loss-of-function experiments, we demonstrate that Id2 is essential for anchorage independent survival (anoikis resistance) in vitro and lung colonization in mice. Importantly, we find that under anchorage independent conditions, E-cadherin loss promotes expression of Id2 in multiple mouse and (organotypic) human models of ILC, an event that is caused by a direct p120-catenin/Kaiso-dependent transcriptional de-repression of the canonical Kaiso binding sequence TCCTGCNA. Conversely, stable inducible restoration of E-cadherin expression in the ILC cell line SUM44PE inhibits Id2 expression and anoikis resistance. We show evidence that Id2 accumulates in the cytosol, where it induces a sustained and CDK4/6-dependent G0/G1 cell cycle arrest through interaction with hypo-phosphorylated Rb. Finally, we find that Id2 is indeed enriched in ILC when compared to other breast cancers, and confirm cytosolic Id2 protein expression in primary ILC samples. In sum, we have linked mutational inactivation of E-cadherin to direct inhibition of cell cycle progression. Our work indicates that loss of E-cadherin and subsequent expression of Id2 drive indolence and dissemination of ILC. As such, E-cadherin and Id2 are promising candidates to stratify low and intermediate grade invasive breast cancers for the use of clinical cell cycle intervention drugs..
Love, S.B.
Cafferty, F.
Snowdon, C.
Carty, K.
Savage, J.
Pallmann, P.
McParland, L.
Brown, L.
Masters, L.
Schiavone, F.
Hague, D.
Townsend, S.
Amos, C.
South, A.
Sturgeon, K.
Langley, R.
Maughan, T.
James, N.
Hall, E.
Kernaghan, S.
Bliss, J.
Turner, N.
Tutt, A.
Yap, C.
Firth, C.
Kong, A.
Mehanna, H.
Watts, C.
Hills, R.
Thomas, I.
Copland, M.
Bell, S.
Sebag-Montefiore, D.
Jones, R.
Parmar, M.K.
Sydes, M.R.
(2022). Practical guidance for running late-phase platform protocols for clinical trials: lessons from experienced UK clinical trials units. Trials,
Vol.23
(1),
p. 757.
show abstract
full text
BACKGROUND: Late-phase platform protocols (including basket, umbrella, multi-arm multi-stage (MAMS), and master protocols) are generally agreed to be more efficient than traditional two-arm clinical trial designs but are not extensively used. We have gathered the experience of running a number of successful platform protocols together to present some operational recommendations. METHODS: Representatives of six UK clinical trials units with experience in running late-phase platform protocols attended a 1-day meeting structured to discuss various practical aspects of running these trials. We report and give guidance on operational aspects which are either harder to implement compared to a traditional late-phase trial or are specific to platform protocols. RESULTS: We present a list of practical recommendations for trialists intending to design and conduct late-phase platform protocols. Our recommendations cover the entire life cycle of a platform trial: from protocol development, obtaining funding, and trial set-up, to a wide range of operational and regulatory aspects such as staffing, oversight, data handling, and data management, to the reporting of results, with a particular focus on communication with trial participants and stakeholders as well as public and patient involvement. DISCUSSION: Platform protocols enable many questions to be answered efficiently to the benefit of patients. Our practical lessons from running platform trials will support trial teams in learning how to run these trials more effectively and efficiently..
Telli, M.L.
Tolaney, S.M.
Shapiro, G.I.
Middleton, M.
Lord, S.R.
Arkenau, H.T.
Tutt, A.
Abramson, V.
Dean, E.
Haddad, T.C.
Wesolowski, R.
Ferrer-Playan, J.
Goddemeier, T.
Grombacher, T.
Dong, J.
Fleuranceau-Morel, P.
Diaz-Padilla, I.
Plummer, R.
(2022). Phase 1b study of berzosertib and cisplatin in patients with advanced triple-negative breast cancer. Npj breast cancer,
Vol.8
(1),
p. 45.
show abstract
full text
Platinum derivatives are commonly used for the treatment of patients with metastatic triple-negative breast cancer (TNBC). However, resistance often develops, leading to treatment failure. This expansion cohort (part C2) of the previously reported phase 1b trial (NCT02157792) is based on the recommended phase 2 dose of the combination of the ataxia-telangiectasia and Rad3-related (ATR) inhibitor berzosertib and cisplatin observed in patients with advanced solid tumors, including TNBC. Forty-seven patients aged ≥18 years with advanced TNBC received cisplatin (75 mg/m2; day 1) and berzosertib (140 mg/m2; days 2 and 9), in 21-day cycles. Berzosertib was well tolerated, with a similar toxicity profile to that reported previously for this combination. The overall response rate (90% confidence interval) was 23.4% (13.7, 35.8). No relevant associations were observed between response and gene alterations. Further studies combining ATR inhibitors with platinum compounds may be warranted in highly selected patient populations..
Tarantino, D.
Walker, C.
Weekes, D.
Pemberton, H.
Davidson, K.
Torga, G.
Frankum, J.
Mendes-Pereira, A.M.
Prince, C.
Ferro, R.
Brough, R.
Pettitt, S.J.
Lord, C.J.
Grigoriadis, A.
Nj Tutt, A.
(2022). Functional screening reveals HORMAD1-driven gene dependencies associated with translesion synthesis and replication stress tolerance. Oncogene,
Vol.41
(32),
pp. 3969-3977.
show abstract
full text
HORMAD1 expression is usually restricted to germline cells, but it becomes mis-expressed in epithelial cells in ~60% of triple-negative breast cancers (TNBCs), where it is associated with elevated genomic instability (1). HORMAD1 expression in TNBC is bimodal with HORMAD1-positive TNBC representing a biologically distinct disease group. Identification of HORMAD1-driven genetic dependencies may uncover novel therapies for this disease group. To study HORMAD1-driven genetic dependencies, we generated a SUM159 cell line model with doxycycline-inducible HORMAD1 that replicated genomic instability phenotypes seen in HORMAD1-positive TNBC (1). Using small interfering RNA screens, we identified candidate genes whose depletion selectively inhibited the cellular growth of HORMAD1-expressing cells. We validated five genes (ATR, BRIP1, POLH, TDP1 and XRCC1), depletion of which led to reduced cellular growth or clonogenic survival in cells expressing HORMAD1. In addition to the translesion synthesis (TLS) polymerase POLH, we identified a HORMAD1-driven dependency upon additional TLS polymerases, namely POLK, REV1, REV3L and REV7. Our data confirms that out-of-context somatic expression of HORMAD1 can lead to genomic instability and reveals that HORMAD1 expression induces dependencies upon replication stress tolerance pathways, such as translesion synthesis. Our data also suggest that HORMAD1 expression could be a patient selection biomarker for agents targeting replication stress..
Geyer, C.E.
Garber, J.E.
Gelber, R.D.
Yothers, G.
Taboada, M.
Ross, L.
Rastogi, P.
Cui, K.
Arahmani, A.
Aktan, G.
Armstrong, A.C.
Arnedos, M.
Balmaña, J.
Bergh, J.
Bliss, J.
Delaloge, S.
Domchek, S.M.
Eisen, A.
Elsafy, F.
Fein, L.E.
Fielding, A.
Ford, J.M.
Friedman, S.
Gelmon, K.A.
Gianni, L.
Gnant, M.
Hollingsworth, S.J.
Im, S.-.
Jager, A.
Jóhannsson, Ó.Þ.
Lakhani, S.R.
Janni, W.
Linderholm, B.
Liu, T.-.
Loman, N.
Korde, L.
Loibl, S.
Lucas, P.C.
Marmé, F.
Martinez de Dueñas, E.
McConnell, R.
Phillips, K.-.
Piccart, M.
Rossi, G.
Schmutzler, R.
Senkus, E.
Shao, Z.
Sharma, P.
Singer, C.F.
Španić, T.
Stickeler, E.
Toi, M.
Traina, T.A.
Viale, G.
Zoppoli, G.
Park, Y.H.
Yerushalmi, R.
Yang, H.
Pang, D.
Jung, K.H.
Mailliez, A.
Fan, Z.
Tennevet, I.
Zhang, J.
Nagy, T.
Sonke, G.S.
Sun, Q.
Parton, M.
Colleoni, M.A.
Schmidt, M.
Brufsky, A.M.
Razaq, W.
Kaufman, B.
Cameron, D.
Campbell, C.
Tutt, A.N.
OlympiA Clinical Trial Steering Committee and Investigators,
(2022). Overall survival in the OlympiA phase III trial of adjuvant olaparib in patients with germline pathogenic variants in BRCA1/2 and high-risk, early breast cancer. Ann oncol,
Vol.33
(12),
pp. 1250-1268.
show abstract
full text
BACKGROUND: The randomized, double-blind OlympiA trial compared 1 year of the oral poly(adenosine diphosphate-ribose) polymerase inhibitor, olaparib, to matching placebo as adjuvant therapy for patients with pathogenic or likely pathogenic variants in germline BRCA1 or BRCA2 (gBRCA1/2pv) and high-risk, human epidermal growth factor receptor 2-negative, early breast cancer (EBC). The first pre-specified interim analysis (IA) previously demonstrated statistically significant improvement in invasive disease-free survival (IDFS) and distant disease-free survival (DDFS). The olaparib group had fewer deaths than the placebo group, but the difference did not reach statistical significance for overall survival (OS). We now report the pre-specified second IA of OS with updates of IDFS, DDFS, and safety. PATIENTS AND METHODS: One thousand eight hundred and thirty-six patients were randomly assigned to olaparib or placebo following (neo)adjuvant chemotherapy, surgery, and radiation therapy if indicated. Endocrine therapy was given concurrently with study medication for hormone receptor-positive cancers. Statistical significance for OS at this IA required P < 0.015. RESULTS: With a median follow-up of 3.5 years, the second IA of OS demonstrated significant improvement in the olaparib group relative to the placebo group [hazard ratio 0.68; 98.5% confidence interval (CI) 0.47-0.97; P = 0.009]. Four-year OS was 89.8% in the olaparib group and 86.4% in the placebo group (Δ 3.4%, 95% CI -0.1% to 6.8%). Four-year IDFS for the olaparib group versus placebo group was 82.7% versus 75.4% (Δ 7.3%, 95% CI 3.0% to 11.5%) and 4-year DDFS was 86.5% versus 79.1% (Δ 7.4%, 95% CI 3.6% to 11.3%), respectively. Subset analyses for OS, IDFS, and DDFS demonstrated benefit across major subgroups. No new safety signals were identified including no new cases of acute myeloid leukemia or myelodysplastic syndrome. CONCLUSION: With 3.5 years of median follow-up, OlympiA demonstrates statistically significant improvement in OS with adjuvant olaparib compared with placebo for gBRCA1/2pv-associated EBC and maintained improvements in the previously reported, statistically significant endpoints of IDFS and DDFS with no new safety signals..
Gazinska, P.
Milton, C.
Iacovacci, J.
Ward, J.
Buus, R.
Alaguthurai, T.
Graham, R.
Akarca, A.
Lips, E.
Naidoo, K.
Wesseling, J.
Marafioti, T.
Cheang, M.
Gillett, C.
Wu, Y.
Khan, A.
Melcher, A.
Salgado, R.
Dowsett, M.
Tutt, A.
Roxanis, I.
Haider, S.
Irshad, S.
(2022). Dynamic Changes in the NK-, Neutrophil-, and B-cell Immunophenotypes Relevant in High Metastatic Risk Post Neoadjuvant Chemotherapy-Resistant Early Breast Cancers. Clin cancer res,
Vol.28
(20),
pp. 4494-4508.
show abstract
full text
PURPOSE: To identify potential immune targets in post-neoadjuvant chemotherapy (NAC)-resistant triple-negative breast cancer (TNBC) and ER+HER2- breast cancer disease. EXPERIMENTAL DESIGN: Following pathology review, 153 patients were identified as having residual cancer burden (RCB) II/III disease (TNBC n = 80; ER+HER2-n = 73). Baseline pre-NAC samples were available for evaluation for 32 of 80 TNBC and 36 of 73 ER+HER2- cases. Bright-field hematoxylin and eosin assessment allowed for tumor-infiltrating lymphocyte (TIL) evaluation in all cases. Multiplexed immunofluorescence was used to identify the abundance and distribution of immune cell subsets. Levels of checkpoints including PD-1/PD-L1 expression were also quantified. Findings were then validated using expression profiling of cancer and immune-related genes. Cytometry by time-of-flight characterized the dynamic changes in circulating immune cells with NAC. RESULTS: RCB II/III TNBC and ER+HER2- breast cancer were immunologically "cold" at baseline and end of NAC. Although the distribution of immune cell subsets across subtypes was similar, the mRNA expression profiles were both subtype- and chemotherapy-specific. TNBC RCB II/III disease was enriched with genes related to neutrophil degranulation, and displayed strong interplay across immune and cancer pathways. We observed similarities in the dynamic changes in B-cell biology following NAC irrespective of subtype. However, NAC induced changes in the local and circulating tumor immune microenvironment (TIME) that varied by subtype and response. Specifically, in TNBC residual disease, we observed downregulation of stimulatory (CD40/OX40L) and inhibitory (PD-L1/PD-1) receptor expression and an increase in NK cell populations (especially non-cytolytic, exhausted CD56dimCD16-) within both the local TIME and peripheral white cell populations. CONCLUSIONS: This study identifies several potential immunologic pathways in residual disease, which may be targeted to benefit high-risk patients..
Fitzpatrick, A.
Iravani, M.
Mills, A.
Childs, L.
Alaguthurai, T.
Clifford, A.
Garcia-Murillas, I.
Van Laere, S.
Dirix, L.
Harries, M.
Okines, A.
Turner, N.C.
Haider, S.
Tutt, A.N.
Isacke, C.M.
(2022). Assessing CSF ctDNA to Improve Diagnostic Accuracy and Therapeutic Monitoring in Breast Cancer Leptomeningeal Metastasis. Clin cancer res,
Vol.28
(6),
pp. 1180-1191.
show abstract
full text
PURPOSE: Cerebrospinal fluid (CSF) cytology is the gold standard diagnostic test for breast cancer leptomeningeal metastasis (BCLM), but has impaired sensitivity, often necessitating repeated lumbar puncture to confirm or refute diagnosis. Further, there is no quantitative response tool to assess response or progression during BCLM treatment. EXPERIMENTAL DESIGN: Facing the challenge of working with small-volume samples and the lack of common recurrent mutations in breast cancers, cell-free DNA was extracted from the CSF and plasma of patients undergoing investigation for BCLM (n = 30). ctDNA fraction was assessed by ultra-low-pass whole genome sequencing (ulpWGS), which does not require prior tumor sequencing. RESULTS: In this proof-of-concept study, ctDNA was detected (fraction ≥0.10) in the CSF of all 24 patients with BCLM+ (median ctDNA fraction, 0.57), regardless of negative cytology or borderline MRI imaging, whereas CSF ctDNA was not detected in the six patients with BCLM- (median ctDNA fraction 0.03, P < 0.0001). Plasma ctDNA was only detected in patients with extracranial disease progression or who had previously received whole brain radiotherapy. ctDNA fraction was highly concordant with mutant allele fraction measured by tumor mutation-specific ddPCR assays (r = 0.852; P < 0.0001). During intrathecal treatment, serial monitoring (n = 12 patients) showed that suppression of CSF ctDNA fraction was associated with longer BCLM survival (P = 0.034), and rising ctDNA fraction was detectable up to 12 weeks before clinical progression. CONCLUSIONS: Measuring ctDNA fraction by ulpWGS is a quantitative marker demonstrating potential for timely and accurate BCLM diagnosis and therapy response monitoring, with the ultimate aim to improve management of this poor-prognosis patient group..
Sipos, O.
Tovey, H.
Quist, J.
Haider, S.
Nowinski, S.
Gazinska, P.
Kernaghan, S.
Toms, C.
Maguire, S.
Orr, N.
Linn, S.C.
Owen, J.
Gillett, C.
Pinder, S.E.
Bliss, J.M.
Tutt, A.
Cheang, M.C.
Grigoriadis, A.
(2021). Assessment of structural chromosomal instability phenotypes as biomarkers of carboplatin response in triple negative breast cancer: the TNT trial. Ann oncol,
Vol.32
(1),
pp. 58-65.
show abstract
full text
BACKGROUND: In the TNT trial of triple negative breast cancer (NCT00532727), germline BRCA1/2 mutations were present in 28% of carboplatin responders. We assessed quantitative measures of structural chromosomal instability (CIN) to identify a wider patient subgroup within TNT with preferential benefit from carboplatin over docetaxel. PATIENTS AND METHODS: Copy number aberrations (CNAs) were established from 135 formalin-fixed paraffin-embedded primary carcinomas using Illumina OmniExpress SNP-arrays. Seven published [allelic imbalanced CNA (AiCNA); allelic balanced CNA (AbCNA); copy number neutral loss of heterozygosity (CnLOH); number of telomeric allelic imbalances (NtAI); BRCA1-like status; percentage of genome altered (PGA); homologous recombination deficiency (HRD) scores] and two novel [Shannon diversity index (SI); high-level amplifications (HLAMP)] CIN-measurements were derived. HLAMP was defined based on the presence of at least one of the top 5% amplified cytobands located on 1q, 8q and 10p. Continuous CIN-measurements were divided into tertiles. All nine CIN-measurements were used to analyse objective response rate (ORR) and progression-free survival (PFS). RESULTS: Patients with tumours without HLAMP had a numerically higher ORR and significantly longer PFS in the carboplatin (C) than in the docetaxel (D) arm [56% (C) versus 29% (D), PHLAMP,quiet = 0.085; PFS 6.1 months (C) versus 4.1 months (D), Pinteraction/HLAMP = 0.047]. In the carboplatin arm, patients with tumours showing intermediate telomeric NtAI and AiCNA had higher ORR [54% (C) versus 20% (D), PNtAI,intermediate = 0.03; 62% (C) versus 33% (D), PAiCNA,intermediate = 0.076]. Patients with high AiCNA and PGA had shorter PFS in the carboplatin arm [3.4 months (high) versus 5.7 months (low/intermediate); and 3.8 months (high) versus 5.6 months (low/intermediate), respectively; Pinteraction/AiCNA = 0.027, Padj.interaction/AiCNA = 0.125 and Pinteraction/PGA = 0.053, Padj.interaction/PGA = 0.176], whilst no difference was observed in the docetaxel arm. CONCLUSIONS: Patients with tumours lacking HLAMP and demonstrating intermediate CIN-measurements formed a subgroup benefitting from carboplatin relative to docetaxel treatment within the TNT trial. This suggests a complex and paradoxical relationship between the extent of genomic instability in primary tumours and treatment response in the metastatic setting..
Stevenson, J.
Barrow-McGee, R.
Yu, L.
Paul, A.
Mansfield, D.
Owen, J.
Woodman, N.
Natrajan, R.
Haider, S.
Gillett, C.
Tutt, A.
Pinder, S.E.
Choudary, J.
Naidoo, K.
(2021). Proteomics of REPLICANT perfusate detects changes in the metastatic lymph node microenvironment. Npj breast cancer,
Vol.7
(1),
p. 24.
show abstract
full text
In breast cancer (BC), detecting low volumes of axillary lymph node (ALN) metastasis pre-operatively is difficult and novel biomarkers are needed. We recently showed that patient-derived ALNs can be sustained ex-vivo using normothermic perfusion. We now compare reactive (tumour-free; n = 5) and macrometastatic (containing tumour deposits >2 mm; n = 4) ALNs by combining whole section multiplex immunofluorescence with TMT-labelled LC-MS/MS of the circulating perfusate. Macrometastases contained significantly fewer B cells and T cells (CD4+/CD8+/regulatory) than reactive nodes (p = 0.02). Similarly, pathway analysis of the perfusate proteome (119/1453 proteins significantly differentially expressed) showed that immune function was diminished in macrometastases in favour of 'extracellular matrix degradation'; only 'neutrophil degranulation' was preserved. Qualitative comparison of the perfusate proteome to that of node-positive pancreatic and prostatic adenocarcinoma also highlighted 'neutrophil degranulation' as a contributing factor to nodal metastasis. Thus, metastasis-induced changes in the REPLICANT perfusate proteome are detectable, and could facilitate biomarker discovery..
Bajrami, I.
Walker, C.
Krastev, D.B.
Weekes, D.
Song, F.
Wicks, A.J.
Alexander, J.
Haider, S.
Brough, R.
Pettitt, S.J.
Tutt, A.N.
Lord, C.J.
(2021). Sirtuin inhibition is synthetic lethal with BRCA1 or BRCA2 deficiency. Commun biol,
Vol.4
(1),
p. 1270.
show abstract
full text
PARP enzymes utilise NAD+ as a co-substrate for their enzymatic activity. Inhibition of PARP1 is synthetic lethal with defects in either BRCA1 or BRCA2. In order to assess whether other genes implicated in NAD+ metabolism were synthetic lethal with BRCA1 or BRCA2 gene defects, we carried out a genetic screen, which identified a synthetic lethality between BRCA1 and genetic inhibition of either of two sirtuin (SIRT) enzymes, SIRT1 or SIRT6. This synthetic lethal interaction was replicated using small-molecule SIRT inhibitors and was associated with replication stress and increased cellular PARylation, in contrast to the decreased PARylation associated with BRCA-gene/PARP inhibitor synthetic lethality. SIRT/BRCA1 synthetic lethality was reversed by genetic ablation of either PARP1 or the histone PARylation factor-coding gene HPF1, implicating PARP1/HPF1-mediated serine ADP-ribosylation as part of the mechanistic basis of this synthetic lethal effect. These observations suggest that PARP1/HPF1-mediated serine ADP-ribosylation, when driven by SIRT inhibition, can inadvertently inhibit the growth of BRCA-gene mutant cells..
Early Breast Cancer Trialists’ Collaborative group (EBCTCG),
(2021). Trastuzumab for early-stage, HER2-positive breast cancer: a meta-analysis of 13 864 women in seven randomised trials. Lancet oncol,
Vol.22
(8),
pp. 1139-1150.
show abstract
full text
BACKGROUND: Trastuzumab targets the extracellular domain of the HER2 protein. Adding trastuzumab to chemotherapy for patients with early-stage, HER2-positive breast cancer reduces the risk of recurrence and death, but is associated with cardiac toxicity. We investigated the long-term benefits and risks of adjuvant trastuzumab on breast cancer recurrence and cause-specific mortality. METHODS: We did a collaborative meta-analysis of individual patient data from randomised trials assessing chemotherapy plus trastuzumab versus the same chemotherapy alone. Randomised trials that enrolled women with node-negative or node-positive, operable breast cancer were included. We collected individual patient-level data on baseline characteristics, dates and sites of first distant breast cancer recurrence and any previous local recurrence or second primary cancer, and the date and underlying cause of death. Primary outcomes were breast cancer recurrence, breast cancer mortality, death without recurrence, and all-cause mortality. Standard intention-to-treat log-rank analyses, stratified by age, nodal status, oestrogen receptor (ER) status, and trial yielded first-event rate ratios (RRs). FINDINGS: Seven randomised trials met the inclusion criteria, and included 13 864 patients enrolled between February, 2000, and December, 2005. Mean scheduled treatment duration was 14·4 months and median follow-up was 10·7 years (IQR 9·5 to 11·9). The risks of breast cancer recurrence (RR 0·66, 95% CI 0·62 to 0·71; p<0·0001) and death from breast cancer (0·67, 0·61 to 0·73; p<0·0001) were lower with trastuzumab plus chemotherapy than with chemotherapy alone. Absolute 10-year recurrence risk was reduced by 9·0% (95% CI 7·4 to 10·7; p<0·0001) and 10-year breast cancer mortality was reduced by 6·4% (4·9 to 7·8; p<0·0001), with a 6·5% reduction (5·0 to 8·0; p<0·0001) in all-cause mortality, and no increase in death without recurrence (0·4%, -0·3 to 1·1; p=0·35). The proportional reduction in recurrence was largest in years 0-1 after randomisation (0·53, 99% CI 0·46 to 0·61), with benefits persisting through years 2-4 (0·73, 0·62 to 0·85) and 5-9 (0·80, 0·64 to 1·01), and little follow-up beyond year 10. Proportional recurrence reductions were similar irrespective of recorded patient and tumour characteristics, including ER status. The more high risk the tumour, the larger the absolute reductions in 5-year recurrence (eg, 5·7% [95% CI 3·1 to 8·3], 6·8% [4·7 to 9·0], and 10·7% [7·7 to 13·6] in N0, N1-3, and N4+ disease). INTERPRETATION: Adding trastuzumab to chemotherapy for early-stage, HER2-positive breast cancer reduces recurrence of, and mortality from, breast cancer by a third, with worthwhile proportional reductions irrespective of recorded patient and tumour characteristics. FUNDING: Cancer Research UK, UK Medical Research Council..
Harvey-Jones, E.
Vinas Villaro, G.
Tutt, A.
(2021). New Roles of Poly(ADP-Ribose) Polymerase Inhibitors in the Treatment of Breast Cancer. Cancer j,
Vol.27
(6),
pp. 441-456.
show abstract
full text
Since the proof of concept of synthetic lethality between poly(ADP-ribose) polymerase inhibition and loss of BRCA1/2 homologous recombination (HR) function in preclinical models and early phase clinical trials, poly(ADP-ribose) polymerase inhibitors (PARPi) are increasing part of standard-of-care treatment for advanced breast cancers with BRCA gene mutations. The field has also recently seen benefits for PARPi in early breast cancer in those with germline BRCA1 and BRCA2 pathogenic mutations, and signals that synthetic lethal affects may occur in tumors with deficiencies in HR caused by germline, somatic, or epigenetic dysregulation of a number of HR genes. Despite the evidence of the synthetic lethal effects of PARPi, they are not always effective in HR defective cancers, and as they become part of standard of care in breast cancer, the study of prevalence of distinct mechanisms of resistance to PARPi and cross-resistance with other DNA-damaging agents such as platinum in breast cancer will be important and may inform therapy choices..
Peck, B.
Bland, P.
Mavrommati, I.
Muirhead, G.
Cottom, H.
Wai, P.T.
Maguire, S.L.
Barker, H.E.
Morrison, E.
Kriplani, D.
Yu, L.
Gibson, A.
Falgari, G.
Brennan, K.
Farnie, G.
Buus, R.
Marlow, R.
Novo, D.
Knight, E.
Guppy, N.
Kolarevic, D.
Susnjar, S.
Milijic, N.M.
Naidoo, K.
Gazinska, P.
Roxanis, I.
Pancholi, S.
Martin, L.-.
Holgersen, E.M.
Cheang, M.C.
Noor, F.
Postel-Vinay, S.
Quinn, G.
McDade, S.
Krasny, L.
Huang, P.
Daley, F.
Wallberg, F.
Choudhary, J.S.
Haider, S.
Tutt, A.N.
Natrajan, R.
(2021). 3D Functional Genomics Screens Identify CREBBP as a Targetable Driver in Aggressive Triple-Negative Breast Cancer. Cancer res,
Vol.81
(4),
pp. 847-859.
show abstract
full text
Triple-negative breast cancers (TNBC) are resistant to standard-of-care chemotherapy and lack known targetable driver gene alterations. Identification of novel drivers could aid the discovery of new treatment strategies for this hard-to-treat patient population, yet studies using high-throughput and accurate models to define the functions of driver genes in TNBC to date have been limited. Here, we employed unbiased functional genomics screening of the 200 most frequently mutated genes in breast cancer, using spheroid cultures to model in vivo-like conditions, and identified the histone acetyltransferase CREBBP as a novel tumor suppressor in TNBC. CREBBP protein expression in patient tumor samples was absent in 8% of TNBCs and at a high frequency in other tumors, including squamous lung cancer, where CREBBP-inactivating mutations are common. In TNBC, CREBBP alterations were associated with higher genomic heterogeneity and poorer patient survival and resulted in upregulation and dependency on a FOXM1 proliferative program. Targeting FOXM1-driven proliferation indirectly with clinical CDK4/6 inhibitors (CDK4/6i) selectively impaired growth in spheroids, cell line xenografts, and patient-derived models from multiple tumor types with CREBBP mutations or loss of protein expression. In conclusion, we have identified CREBBP as a novel driver in aggressive TNBC and identified an associated genetic vulnerability in tumor cells with alterations in CREBBP and provide a preclinical rationale for assessing CREBBP alterations as a biomarker of CDK4/6i response in a new patient population. SIGNIFICANCE: This study demonstrates that CREBBP genomic alterations drive aggressive TNBC, lung cancer, and lymphomas and may be selectively treated with clinical CDK4/6 inhibitors..
Harris, R.J.
Cheung, A.
Ng, J.C.
Laddach, R.
Chenoweth, A.M.
Crescioli, S.
Fittall, M.
Dominguez-Rodriguez, D.
Roberts, J.
Levi, D.
Liu, F.
Alberts, E.
Quist, J.
Santaolalla, A.
Pinder, S.E.
Gillett, C.
Hammar, N.
Irshad, S.
Van Hemelrijck, M.
Dunn-Walters, D.K.
Fraternali, F.
Spicer, J.F.
Lacy, K.E.
Tsoka, S.
Grigoriadis, A.
Tutt, A.N.
Karagiannis, S.N.
(2021). Tumor-Infiltrating B Lymphocyte Profiling Identifies IgG-Biased, Clonally Expanded Prognostic Phenotypes in Triple-Negative Breast Cancer. Cancer res,
Vol.81
(16),
pp. 4290-4304.
show abstract
full text
In breast cancer, humoral immune responses may contribute to clinical outcomes, especially in more immunogenic subtypes. Here, we investigated B lymphocyte subsets, immunoglobulin expression, and clonal features in breast tumors, focusing on aggressive triple-negative breast cancers (TNBC). In samples from patients with TNBC and healthy volunteers, circulating and tumor-infiltrating B lymphocytes (TIL-B) were evaluated. CD20+CD27+IgD- isotype-switched B lymphocytes were increased in tumors, compared with matched blood. TIL-B frequently formed stromal clusters with T lymphocytes and engaged in bidirectional functional cross-talk, consistent with gene signatures associated with lymphoid assembly, costimulation, cytokine-cytokine receptor interactions, cytotoxic T-cell activation, and T-cell-dependent B-cell activation. TIL-B-upregulated B-cell receptor (BCR) pathway molecules FOS and JUN, germinal center chemokine regulator RGS1, activation marker CD69, and TNFα signal transduction via NFκB, suggesting BCR-immune complex formation. Expression of genes associated with B lymphocyte recruitment and lymphoid assembly, including CXCL13, CXCR4, and DC-LAMP, was elevated in TNBC compared with other subtypes and normal breast. TIL-B-rich tumors showed expansion of IgG but not IgA isotypes, and IgG isotype switching positively associated with survival outcomes in TNBC. Clonal expansion was biased toward IgG, showing expansive clonal families with specific variable region gene combinations and narrow repertoires. Stronger positive selection pressure was present in the complementarity determining regions of IgG compared with their clonally related IgA in tumor samples. Overall, class-switched B lymphocyte lineage traits were conspicuous in TNBC, associated with improved clinical outcomes, and conferred IgG-biased, clonally expanded, and likely antigen-driven humoral responses. SIGNIFICANCE: Tumor-infiltrating B lymphocytes assemble in clusters, undergoing B-cell receptor-driven activation, proliferation, and isotype switching. Clonally expanded, IgG isotype-biased humoral immunity associates with favorable prognosis primarily in triple-negative breast cancers..
Turner, N.C.
Balmaña, J.
Poncet, C.
Goulioti, T.
Tryfonidis, K.
Honkoop, A.H.
Zoppoli, G.
Razis, E.
Johannsson, O.T.
Colleoni, M.
Tutt, A.N.
Audeh, W.
Ignatiadis, M.
Mailliez, A.
Trédan, O.
Musolino, A.
Vuylsteke, P.
Juan-Fita, M.J.
Macpherson, I.R.
Kaufman, B.
Manso, L.
Goldstein, L.J.
Ellard, S.L.
Láng, I.
Jen, K.Y.
Adam, V.
Litière, S.
Erban, J.
Cameron, D.A.
BRAVO Steering Committee and the BRAVO investigators,
(2021). Niraparib for Advanced Breast Cancer with Germline BRCA1 and BRCA2 Mutations: the EORTC 1307-BCG/BIG5-13/TESARO PR-30-50-10-C BRAVO Study. Clin cancer res,
Vol.27
(20),
pp. 5482-5491.
show abstract
full text
PURPOSE: To investigate the activity of niraparib in patients with germline-mutated BRCA1/2 (gBRCAm) advanced breast cancer. PATIENTS AND METHODS: BRAVO was a randomized, open-label phase III trial. Eligible patients had gBRCAm and HER2-negative advanced breast cancer previously treated with ≤2 prior lines of chemotherapy for advanced breast cancer or had relapsed within 12 months of adjuvant chemotherapy, and were randomized 2:1 between niraparib and physician's choice chemotherapy (PC; monotherapy with eribulin, capecitabine, vinorelbine, or gemcitabine). Patients with hormone receptor-positive tumors had to have received ≥1 line of endocrine therapy and progressed during this treatment in the metastatic setting or relapsed within 1 year of (neo)adjuvant treatment. The primary endpoint was centrally assessed progression-free survival (PFS). Secondary endpoints included overall survival (OS), PFS by local assessment (local-PFS), objective response rate (ORR), and safety. RESULTS: After the pre-planned interim analysis, recruitment was halted on the basis of futility, noting a high degree of discordance between local and central PFS assessment in the PC arm that resulted in informative censoring. At the final analysis (median follow-up, 19.9 months), median centrally assessed PFS was 4.1 months in the niraparib arm (n = 141) versus 3.1 months in the PC arm [n = 74; hazard ratio (HR), 0.96; 95% confidence interval (CI), 0.65-1.44; P = 0.86]. HRs for OS and local-PFS were 0.95 (95% CI, 0.63-1.42) and 0.65 (95% CI, 0.46-0.93), respectively. ORR was 35% (95% CI, 26-45) with niraparib and 31% (95% CI, 19-46) in the PC arm. CONCLUSIONS: Informative censoring in the control arm prevented accurate assessment of the trial hypothesis, although there was clear evidence of niraparib's activity in this patient population..
Zatreanu, D.
Robinson, H.M.
Alkhatib, O.
Boursier, M.
Finch, H.
Geo, L.
Grande, D.
Grinkevich, V.
Heald, R.A.
Langdon, S.
Majithiya, J.
McWhirter, C.
Martin, N.M.
Moore, S.
Neves, J.
Rajendra, E.
Ranzani, M.
Schaedler, T.
Stockley, M.
Wiggins, K.
Brough, R.
Sridhar, S.
Gulati, A.
Shao, N.
Badder, L.M.
Novo, D.
Knight, E.G.
Marlow, R.
Haider, S.
Callen, E.
Hewitt, G.
Schimmel, J.
Prevo, R.
Alli, C.
Ferdinand, A.
Bell, C.
Blencowe, P.
Bot, C.
Calder, M.
Charles, M.
Curry, J.
Ekwuru, T.
Ewings, K.
Krajewski, W.
MacDonald, E.
McCarron, H.
Pang, L.
Pedder, C.
Rigoreau, L.
Swarbrick, M.
Wheatley, E.
Willis, S.
Wong, A.C.
Nussenzweig, A.
Tijsterman, M.
Tutt, A.
Boulton, S.J.
Higgins, G.S.
Pettitt, S.J.
Smith, G.C.
Lord, C.J.
(2021). Polθ inhibitors elicit BRCA-gene synthetic lethality and target PARP inhibitor resistance. Nat commun,
Vol.12
(1),
p. 3636.
show abstract
full text
To identify approaches to target DNA repair vulnerabilities in cancer, we discovered nanomolar potent, selective, low molecular weight (MW), allosteric inhibitors of the polymerase function of DNA polymerase Polθ, including ART558. ART558 inhibits the major Polθ-mediated DNA repair process, Theta-Mediated End Joining, without targeting Non-Homologous End Joining. In addition, ART558 elicits DNA damage and synthetic lethality in BRCA1- or BRCA2-mutant tumour cells and enhances the effects of a PARP inhibitor. Genetic perturbation screening revealed that defects in the 53BP1/Shieldin complex, which cause PARP inhibitor resistance, result in in vitro and in vivo sensitivity to small molecule Polθ polymerase inhibitors. Mechanistically, ART558 increases biomarkers of single-stranded DNA and synthetic lethality in 53BP1-defective cells whilst the inhibition of DNA nucleases that promote end-resection reversed these effects, implicating these in the synthetic lethal mechanism-of-action. Taken together, these observations describe a drug class that elicits BRCA-gene synthetic lethality and PARP inhibitor synergy, as well as targeting a biomarker-defined mechanism of PARPi-resistance..
Beatson, R.E.
Parente-Pereira, A.C.
Halim, L.
Cozzetto, D.
Hull, C.
Whilding, L.M.
Martinez, O.
Taylor, C.A.
Obajdin, J.
Luu Hoang, K.N.
Draper, B.
Iqbal, A.
Hardiman, T.
Zabinski, T.
Man, F.
de Rosales, R.T.
Xie, J.
Aswad, F.
Achkova, D.
Joseph, C.-.
Ciprut, S.
Adami, A.
Roider, H.G.
Hess-Stumpp, H.
Győrffy, B.
Quist, J.
Grigoriadis, A.
Sommer, A.
Tutt, A.N.
Davies, D.M.
Maher, J.
(2021). TGF-β1 potentiates Vγ9Vδ2 T cell adoptive immunotherapy of cancer. Cell rep med,
Vol.2
(12),
p. 100473.
show abstract
full text
Despite its role in cancer surveillance, adoptive immunotherapy using γδ T cells has achieved limited efficacy. To enhance trafficking to bone marrow, circulating Vγ9Vδ2 T cells are expanded in serum-free medium containing TGF-β1 and IL-2 (γδ[T2] cells) or medium containing IL-2 alone (γδ[2] cells, as the control). Unexpectedly, the yield and viability of γδ[T2] cells are also increased by TGF-β1, when compared to γδ[2] controls. γδ[T2] cells are less differentiated and yet display increased cytolytic activity, cytokine release, and antitumor activity in several leukemic and solid tumor models. Efficacy is further enhanced by cancer cell sensitization using aminobisphosphonates or Ara-C. A number of contributory effects of TGF-β are described, including prostaglandin E2 receptor downmodulation, TGF-β insensitivity, and upregulated integrin activity. Biological relevance is supported by the identification of a favorable γδ[T2] signature in acute myeloid leukemia (AML). Given their enhanced therapeutic activity and compatibility with allogeneic use, γδ[T2] cells warrant evaluation in cancer immunotherapy..
Tutt, A.N.
Garber, J.E.
Kaufman, B.
Viale, G.
Fumagalli, D.
Rastogi, P.
Gelber, R.D.
de Azambuja, E.
Fielding, A.
Balmaña, J.
Domchek, S.M.
Gelmon, K.A.
Hollingsworth, S.J.
Korde, L.A.
Linderholm, B.
Bandos, H.
Senkus, E.
Suga, J.M.
Shao, Z.
Pippas, A.W.
Nowecki, Z.
Huzarski, T.
Ganz, P.A.
Lucas, P.C.
Baker, N.
Loibl, S.
McConnell, R.
Piccart, M.
Schmutzler, R.
Steger, G.G.
Costantino, J.P.
Arahmani, A.
Wolmark, N.
McFadden, E.
Karantza, V.
Lakhani, S.R.
Yothers, G.
Campbell, C.
Geyer, C.E.
OlympiA Clinical Trial Steering Committee and Investigators,
(2021). Adjuvant Olaparib for Patients with BRCA1- or BRCA2-Mutated Breast Cancer. N engl j med,
Vol.384
(25),
pp. 2394-2405.
show abstract
full text
BACKGROUND: Poly(adenosine diphosphate-ribose) polymerase inhibitors target cancers with defects in homologous recombination repair by synthetic lethality. New therapies are needed to reduce recurrence in patients with BRCA1 or BRCA2 germline mutation-associated early breast cancer. METHODS: We conducted a phase 3, double-blind, randomized trial involving patients with human epidermal growth factor receptor 2 (HER2)-negative early breast cancer with BRCA1 or BRCA2 germline pathogenic or likely pathogenic variants and high-risk clinicopathological factors who had received local treatment and neoadjuvant or adjuvant chemotherapy. Patients were randomly assigned (in a 1:1 ratio) to 1 year of oral olaparib or placebo. The primary end point was invasive disease-free survival. RESULTS: A total of 1836 patients underwent randomization. At a prespecified event-driven interim analysis with a median follow-up of 2.5 years, the 3-year invasive disease-free survival was 85.9% in the olaparib group and 77.1% in the placebo group (difference, 8.8 percentage points; 95% confidence interval [CI], 4.5 to 13.0; hazard ratio for invasive disease or death, 0.58; 99.5% CI, 0.41 to 0.82; P<0.001). The 3-year distant disease-free survival was 87.5% in the olaparib group and 80.4% in the placebo group (difference, 7.1 percentage points; 95% CI, 3.0 to 11.1; hazard ratio for distant disease or death, 0.57; 99.5% CI, 0.39 to 0.83; P<0.001). Olaparib was associated with fewer deaths than placebo (59 and 86, respectively) (hazard ratio, 0.68; 99% CI, 0.44 to 1.05; P = 0.02); however, the between-group difference was not significant at an interim-analysis boundary of a P value of less than 0.01. Safety data were consistent with known side effects of olaparib, with no excess serious adverse events or adverse events of special interest. CONCLUSIONS: Among patients with high-risk, HER2-negative early breast cancer and germline BRCA1 or BRCA2 pathogenic or likely pathogenic variants, adjuvant olaparib after completion of local treatment and neoadjuvant or adjuvant chemotherapy was associated with significantly longer survival free of invasive or distant disease than was placebo. Olaparib had limited effects on global patient-reported quality of life. (Funded by the National Cancer Institute and AstraZeneca; OlympiA ClinicalTrials.gov number, NCT02032823.)..
ICGC/TCGA Pan-Cancer Analysis of Whole Genomes Consortium,
(2020). Pan-cancer analysis of whole genomes. Nature,
Vol.578
(7793),
pp. 82-93.
show abstract
full text
Cancer is driven by genetic change, and the advent of massively parallel sequencing has enabled systematic documentation of this variation at the whole-genome scale1-3. Here we report the integrative analysis of 2,658 whole-cancer genomes and their matching normal tissues across 38 tumour types from the Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium of the International Cancer Genome Consortium (ICGC) and The Cancer Genome Atlas (TCGA). We describe the generation of the PCAWG resource, facilitated by international data sharing using compute clouds. On average, cancer genomes contained 4-5 driver mutations when combining coding and non-coding genomic elements; however, in around 5% of cases no drivers were identified, suggesting that cancer driver discovery is not yet complete. Chromothripsis, in which many clustered structural variants arise in a single catastrophic event, is frequently an early event in tumour evolution; in acral melanoma, for example, these events precede most somatic point mutations and affect several cancer-associated genes simultaneously. Cancers with abnormal telomere maintenance often originate from tissues with low replicative activity and show several mechanisms of preventing telomere attrition to critical levels. Common and rare germline variants affect patterns of somatic mutation, including point mutations, structural variants and somatic retrotransposition. A collection of papers from the PCAWG Consortium describes non-coding mutations that drive cancer beyond those in the TERT promoter4; identifies new signatures of mutational processes that cause base substitutions, small insertions and deletions and structural variation5,6; analyses timings and patterns of tumour evolution7; describes the diverse transcriptional consequences of somatic mutation on splicing, expression levels, fusion genes and promoter activity8,9; and evaluates a range of more-specialized features of cancer genomes8,10-18..
Barrow-McGee, R.
Procter, J.
Owen, J.
Woodman, N.
Lombardelli, C.
Kothari, A.
Kovacs, T.
Douek, M.
George, S.
Barry, P.A.
Ramsey, K.
Gibson, A.
Buus, R.
Holgersen, E.
Natrajan, R.
Haider, S.
Shattock, M.J.
Gillett, C.
Tutt, A.N.
Pinder, S.E.
Naidoo, K.
(2020). Real-time ex vivo perfusion of human lymph nodes invaded by cancer (REPLICANT): a feasibility study. J pathol,
Vol.250
(3),
pp. 262-274.
show abstract
full text
Understanding how breast cancer (BC) grows in axillary lymph nodes (ALNs), and refining how therapies might halt that process, is clinically important. However, modelling the complex ALN microenvironment is difficult, and no human models exist at present. We harvested ALNs from ten BC patients, and perfused them at 37 °C ex vivo for up to 24 h. Controlled autologous testing showed that ALNs remain viable after 24 h of ex vivo perfusion: haematoxylin and eosin-stained histological appearance and proliferation (by Ki67 immunohistochemistry) did not change significantly over time for any perfused ALN compared with a control from time-point zero. Furthermore, targeted gene expression analysis (NanoString PanCancer IO360 panel) showed that only 21/750 genes were differentially expressed between control and perfused ALNs (|log2 FC| > 1 and q < 0.1): none were involved in apoptosis and metabolism, but rather all 21 genes were involved in immune function and angiogenesis. During perfusion, tissue acid-base balance remained stable. Interestingly, the flow rate increased (p < 0.001) in cancer-replaced (i.e. metastasis occupied more than 90% of the surface area on multiple levels) compared to cancer-free nodes (i.e. nodes with no metastasis on multiple sections). CXCL11 transcripts were significantly more abundant in cancer-replaced nodes, while CXCL12 transcripts were significantly more abundant in cancer-free nodes. These cytokines were also detected in the circulating perfusate. Monoclonal antibodies (nivolumab and trastuzumab) were administered into a further three ALNs to confirm perfusion efficacy. These drugs saturated the nodes; nivolumab even induced cancer cell death. Normothermic ALN perfusion is not only feasible but sustains the tumour microenvironment ex vivo for scientific investigation. This model could facilitate the identification of actionable immuno-oncology targets. © 2019 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland..
Yeow, Z.Y.
Lambrus, B.G.
Marlow, R.
Zhan, K.H.
Durin, M.-.
Evans, L.T.
Scott, P.M.
Phan, T.
Park, E.
Ruiz, L.A.
Moralli, D.
Knight, E.G.
Badder, L.M.
Novo, D.
Haider, S.
Green, C.M.
Tutt, A.N.
Lord, C.J.
Chapman, J.R.
Holland, A.J.
(2020). Targeting TRIM37-driven centrosome dysfunction in 17q23-amplified breast cancer. Nature,
Vol.585
(7825),
pp. 447-452.
show abstract
full text
Genomic instability is a hallmark of cancer, and has a central role in the initiation and development of breast cancer1,2. The success of poly-ADP ribose polymerase inhibitors in the treatment of breast cancers that are deficient in homologous recombination exemplifies the utility of synthetically lethal genetic interactions in the treatment of breast cancers that are driven by genomic instability3. Given that defects in homologous recombination are present in only a subset of breast cancers, there is a need to identify additional driver mechanisms for genomic instability and targeted strategies to exploit these defects in the treatment of cancer. Here we show that centrosome depletion induces synthetic lethality in cancer cells that contain the 17q23 amplicon, a recurrent copy number aberration that defines about 9% of all primary breast cancer tumours and is associated with high levels of genomic instability4-6. Specifically, inhibition of polo-like kinase 4 (PLK4) using small molecules leads to centrosome depletion, which triggers mitotic catastrophe in cells that exhibit amplicon-directed overexpression of TRIM37. To explain this effect, we identify TRIM37 as a negative regulator of centrosomal pericentriolar material. In 17q23-amplified cells that lack centrosomes, increased levels of TRIM37 block the formation of foci that comprise pericentriolar material-these foci are structures with a microtubule-nucleating capacity that are required for successful cell division in the absence of centrosomes. Finally, we find that the overexpression of TRIM37 causes genomic instability by delaying centrosome maturation and separation at mitotic entry, and thereby increases the frequency of mitotic errors. Collectively, these findings highlight TRIM37-dependent genomic instability as a putative driver event in 17q23-amplified breast cancer and provide a rationale for the use of centrosome-targeting therapeutic agents in treating these cancers..
Pettitt, S.J.
Frankum, J.R.
Punta, M.
Lise, S.
Alexander, J.
Chen, Y.
Yap, T.A.
Haider, S.
Tutt, A.N.
Lord, C.J.
(2020). Clinical BRCA1/2 Reversion Analysis Identifies Hotspot Mutations and Predicted Neoantigens Associated with Therapy Resistance. Cancer discov,
Vol.10
(10),
pp. 1475-1488.
show abstract
full text
Reversion mutations in BRCA1 or BRCA2 are associated with resistance to PARP inhibitors and platinum. To better understand the nature of these mutations, we collated, codified, and analyzed more than 300 reversions. This identified reversion "hotspots" and "deserts" in regions encoding the N and C terminus, respectively, of BRCA2, suggesting that pathogenic mutations in these regions may be at higher or lower risk of reversion. Missense and splice-site pathogenic mutations in BRCA1/2 also appeared less likely to revert than truncating mutations. Most reversions were <100 bp deletions. Although many deletions exhibited microhomology, this was not universal, suggesting that multiple DNA-repair processes cause reversion. Finally, we found that many reversions were predicted to encode immunogenic neopeptides, suggesting a route to the treatment of reverted disease. As well as providing a freely available database for the collation of future reversion cases, these observations have implications for how drug resistance might be managed in BRCA-mutant cancers. SIGNIFICANCE: Reversion mutations in BRCA genes are a major cause of clinical platinum and PARP inhibitor resistance. This analysis of all reported clinical reversions suggests that the position of BRCA2 mutations affects the risk of reversion. Many reversions are also predicted to encode tumor neoantigens, providing a potential route to targeting resistance.This article is highlighted in the In This Issue feature, p. 1426..
Kos, Z.
Roblin, E.
Kim, R.S.
Michiels, S.
Gallas, B.D.
Chen, W.
van de Vijver, K.K.
Goel, S.
Adams, S.
Demaria, S.
Viale, G.
Nielsen, T.O.
Badve, S.S.
Symmans, W.F.
Sotiriou, C.
Rimm, D.L.
Hewitt, S.
Denkert, C.
Loibl, S.
Luen, S.J.
Bartlett, J.M.
Savas, P.
Pruneri, G.
Dillon, D.A.
Cheang, M.C.
Tutt, A.
Hall, J.A.
Kok, M.
Horlings, H.M.
Madabhushi, A.
van der Laak, J.
Ciompi, F.
Laenkholm, A.-.
Bellolio, E.
Gruosso, T.
Fox, S.B.
Araya, J.C.
Floris, G.
Hudeček, J.
Voorwerk, L.
Beck, A.H.
Kerner, J.
Larsimont, D.
Declercq, S.
Van den Eynden, G.
Pusztai, L.
Ehinger, A.
Yang, W.
AbdulJabbar, K.
Yuan, Y.
Singh, R.
Hiley, C.
Bakir, M.A.
Lazar, A.J.
Naber, S.
Wienert, S.
Castillo, M.
Curigliano, G.
Dieci, M.-.
André, F.
Swanton, C.
Reis-Filho, J.
Sparano, J.
Balslev, E.
Chen, I.-.
Stovgaard, E.I.
Pogue-Geile, K.
Blenman, K.R.
Penault-Llorca, F.
Schnitt, S.
Lakhani, S.R.
Vincent-Salomon, A.
Rojo, F.
Braybrooke, J.P.
Hanna, M.G.
Soler-Monsó, M.T.
Bethmann, D.
Castaneda, C.A.
Willard-Gallo, K.
Sharma, A.
Lien, H.-.
Fineberg, S.
Thagaard, J.
Comerma, L.
Gonzalez-Ericsson, P.
Brogi, E.
Loi, S.
Saltz, J.
Klaushen, F.
Cooper, L.
Amgad, M.
Moore, D.A.
Salgado, R.
International Immuno-Oncology Biomarker Working Group,
(2020). Pitfalls in assessing stromal tumor infiltrating lymphocytes (sTILs) in breast cancer. Npj breast cancer,
Vol.6
(1),
p. 17.
show abstract
full text
Stromal tumor-infiltrating lymphocytes (sTILs) are important prognostic and predictive biomarkers in triple-negative (TNBC) and HER2-positive breast cancer. Incorporating sTILs into clinical practice necessitates reproducible assessment. Previously developed standardized scoring guidelines have been widely embraced by the clinical and research communities. We evaluated sources of variability in sTIL assessment by pathologists in three previous sTIL ring studies. We identify common challenges and evaluate impact of discrepancies on outcome estimates in early TNBC using a newly-developed prognostic tool. Discordant sTIL assessment is driven by heterogeneity in lymphocyte distribution. Additional factors include: technical slide-related issues; scoring outside the tumor boundary; tumors with minimal assessable stroma; including lymphocytes associated with other structures; and including other inflammatory cells. Small variations in sTIL assessment modestly alter risk estimation in early TNBC but have the potential to affect treatment selection if cutpoints are employed. Scoring and averaging multiple areas, as well as use of reference images, improve consistency of sTIL evaluation. Moreover, to assist in avoiding the pitfalls identified in this analysis, we developed an educational resource available at www.tilsinbreastcancer.org/pitfalls ..
Hudeček, J.
Voorwerk, L.
van Seijen, M.
Nederlof, I.
de Maaker, M.
van den Berg, J.
van de Vijver, K.K.
Sikorska, K.
Adams, S.
Demaria, S.
Viale, G.
Nielsen, T.O.
Badve, S.S.
Michiels, S.
Symmans, W.F.
Sotiriou, C.
Rimm, D.L.
Hewitt, S.M.
Denkert, C.
Loibl, S.
Loi, S.
Bartlett, J.M.
Pruneri, G.
Dillon, D.A.
Cheang, M.C.
Tutt, A.
Hall, J.A.
Kos, Z.
Salgado, R.
Kok, M.
Horlings, H.M.
International Immuno-Oncology Biomarker Working Group,
(2020). Application of a risk-management framework for integration of stromal tumor-infiltrating lymphocytes in clinical trials. Npj breast cancer,
Vol.6
(1),
p. 15.
show abstract
full text
Stromal tumor-infiltrating lymphocytes (sTILs) are a potential predictive biomarker for immunotherapy response in metastatic triple-negative breast cancer (TNBC). To incorporate sTILs into clinical trials and diagnostics, reliable assessment is essential. In this review, we propose a new concept, namely the implementation of a risk-management framework that enables the use of sTILs as a stratification factor in clinical trials. We present the design of a biomarker risk-mitigation workflow that can be applied to any biomarker incorporation in clinical trials. We demonstrate the implementation of this concept using sTILs as an integral biomarker in a single-center phase II immunotherapy trial for metastatic TNBC (TONIC trial, NCT02499367), using this workflow to mitigate risks of suboptimal inclusion of sTILs in this specific trial. In this review, we demonstrate that a web-based scoring platform can mitigate potential risk factors when including sTILs in clinical trials, and we argue that this framework can be applied for any future biomarker-driven clinical trial setting..
Chopra, N.
Tovey, H.
Pearson, A.
Cutts, R.
Toms, C.
Proszek, P.
Hubank, M.
Dowsett, M.
Dodson, A.
Daley, F.
Kriplani, D.
Gevensleben, H.
Davies, H.R.
Degasperi, A.
Roylance, R.
Chan, S.
Tutt, A.
Skene, A.
Evans, A.
Bliss, J.M.
Nik-Zainal, S.
Turner, N.C.
(2020). Homologous recombination DNA repair deficiency and PARP inhibition activity in primary triple negative breast cancer. Nat commun,
Vol.11
(1),
p. 2662.
show abstract
full text
Triple negative breast cancer (TNBC) encompasses molecularly different subgroups, with a subgroup harboring evidence of defective homologous recombination (HR) DNA repair. Here, within a phase 2 window clinical trial, RIO trial (EudraCT 2014-003319-12), we investigate the activity of PARP inhibitors in 43 patients with untreated TNBC. The primary end point, decreased Ki67, occured in 12% of TNBC. In secondary end point analyses, HR deficiency was identified in 69% of TNBC with the mutational-signature-based HRDetect assay. Cancers with HRDetect mutational signatures of HR deficiency had a functional defect in HR, assessed by impaired RAD51 foci formation on end of treatment biopsy. Following rucaparib treatment there was no association of Ki67 change with HR deficiency. In contrast, early circulating tumor DNA dynamics identified activity of rucaparib, with end of treatment ctDNA levels suppressed by rucaparib in mutation-signature HR-deficient cancers. In ad hoc analysis, rucaparib induced expression of interferon response genes in HR-deficient cancers. The majority of TNBCs have a defect in DNA repair, identifiable by mutational signature analysis, that may be targetable with PARP inhibitors..
Chabanon, R.M.
Muirhead, G.
Krastev, D.B.
Adam, J.
Morel, D.
Garrido, M.
Lamb, A.
Hénon, C.
Dorvault, N.
Rouanne, M.
Marlow, R.
Bajrami, I.
Cardeñosa, M.L.
Konde, A.
Besse, B.
Ashworth, A.
Pettitt, S.J.
Haider, S.
Marabelle, A.
Tutt, A.N.
Soria, J.-.
Lord, C.J.
Postel-Vinay, S.
(2019). PARP inhibition enhances tumor cell-intrinsic immunity in ERCC1-deficient non-small cell lung cancer. J clin invest,
Vol.129
(3),
pp. 1211-1228.
show abstract
full text
The cyclic GMP-AMP synthase/stimulator of IFN genes (cGAS/STING) pathway detects cytosolic DNA to activate innate immune responses. Poly(ADP-ribose) polymerase inhibitors (PARPi) selectively target cancer cells with DNA repair deficiencies such as those caused by BRCA1 mutations or ERCC1 defects. Using isogenic cell lines and patient-derived samples, we showed that ERCC1-defective non-small cell lung cancer (NSCLC) cells exhibit an enhanced type I IFN transcriptomic signature and that low ERCC1 expression correlates with increased lymphocytic infiltration. We demonstrated that clinical PARPi, including olaparib and rucaparib, have cell-autonomous immunomodulatory properties in ERCC1-defective NSCLC and BRCA1-defective triple-negative breast cancer (TNBC) cells. Mechanistically, PARPi generated cytoplasmic chromatin fragments with characteristics of micronuclei; these were found to activate cGAS/STING, downstream type I IFN signaling, and CCL5 secretion. Importantly, these effects were suppressed in PARP1-null TNBC cells, suggesting that this phenotype resulted from an on-target effect of PARPi on PARP1. PARPi also potentiated IFN-γ-induced PD-L1 expression in NSCLC cell lines and in fresh patient tumor cells; this effect was enhanced in ERCC1-deficient contexts. Our data provide a preclinical rationale for using PARPi as immunomodulatory agents in appropriately molecularly selected populations..
Quist, J.
Mirza, H.
Cheang, M.C.
Telli, M.L.
O'Shaughnessy, J.A.
Lord, C.J.
Tutt, A.N.
Grigoriadis, A.
(2019). A Four-gene Decision Tree Signature Classification of Triple-negative Breast Cancer: Implications for Targeted Therapeutics. Mol cancer ther,
Vol.18
(1),
pp. 204-212.
show abstract
full text
The molecular complexity of triple-negative breast cancers (TNBCs) provides a challenge for patient management. We set out to characterize this heterogeneous disease by combining transcriptomics and genomics data, with the aim of revealing convergent pathway dependencies with the potential for treatment intervention. A Bayesian algorithm was used to integrate molecular profiles in two TNBC cohorts, followed by validation using five independent cohorts (n = 1,168), including three clinical trials. A four-gene decision tree signature was identified, which robustly classified TNBCs into six subtypes. All four genes in the signature (EXO1, TP53BP2, FOXM1, and RSU1) are associated with either genomic instability, malignant growth, or treatment response. One of the six subtypes, MC6, encompassed the largest proportion of tumors (∼50%) in early diagnosed TNBCs. In TNBC patients with metastatic disease, the MC6 proportion was reduced to 25%, and was independently associated with a higher response rate to platinum-based chemotherapy. In TNBC cell line data, platinum sensitivity was recapitulated, and a sensitivity to the inhibition of the phosphatase PPM1D was revealed. Molecularly, MC6-TNBCs displayed high levels of telomeric allelic imbalances, enrichment of CD4+ and CD8+ immune signatures, and reduced expression of genes negatively regulating the MAPK signaling pathway. These observations suggest that our integrative classification approach may identify TNBC patients with discernible and theoretically pharmacologically tractable features that merit further studies in prospective trials..
Wu, Y.
Kyle-Cezar, F.
Woolf, R.T.
Naceur-Lombardelli, C.
Owen, J.
Biswas, D.
Lorenc, A.
Vantourout, P.
Gazinska, P.
Grigoriadis, A.
Tutt, A.
Hayday, A.
(2019). An innate-like Vδ1+ γδ T cell compartment in the human breast is associated with remission in triple-negative breast cancer. Sci transl med,
Vol.11
(513).
show abstract
Innate-like tissue-resident γδ T cell compartments capable of protecting against carcinogenesis are well established in mice. Conversely, the degree to which they exist in humans, their potential properties, and their contributions to host benefit are mostly unresolved. Here, we demonstrate that healthy human breast harbors a distinct γδ T cell compartment, primarily expressing T cell receptor (TCR) Vδ1 chains, by comparison to Vδ2 chains that predominate in peripheral blood. Breast-resident Vδ1+ cells were functionally skewed toward cytolysis and IFN-γ production, but not IL-17, which has been linked with inflammatory pathologies. Breast-resident Vδ1+ cells could be activated innately via the NKG2D receptor, whereas neighboring CD8+ αβ T cells required TCR signaling. A comparable population of Vδ1+ cells was found in human breast tumors, and when paired tumor and nonmalignant samples from 11 patients with triple-negative breast cancer were analyzed, progression-free and overall survival correlated with Vδ1+ cell representation, but not with either total γδ T cells or Vδ2+ T cells. As expected, progression-free survival also correlated with αβ TCRs. However, whereas in most cases TCRαβ repertoires focused, typical of antigen-specific responses, this was not observed for Vδ1+ cells, consistent with their innate-like responsiveness. Thus, maximal patient benefit may accrue from the collaboration of innate-like responses mounted by tissue-resident Vδ1+ compartments and adaptive responses mounted by αβ T cells..
Mateo, J.
Lord, C.J.
Serra, V.
Tutt, A.
Balmaña, J.
Castroviejo-Bermejo, M.
Cruz, C.
Oaknin, A.
Kaye, S.B.
de Bono, J.S.
(2019). A decade of clinical development of PARP inhibitors in perspective. Ann oncol,
Vol.30
(9),
pp. 1437-1447.
show abstract
full text
Genomic instability is a hallmark of cancer, and often is the result of altered DNA repair capacities in tumour cells. DNA damage repair defects are common in different cancer types; these alterations can also induce tumour-specific vulnerabilities that can be exploited therapeutically. In 2009, a first-in-man clinical trial of the poly(ADP-ribose) polymerase (PARP) inhibitor olaparib clinically validated the synthetic lethal interaction between inhibition of PARP1, a key sensor of DNA damage, and BRCA1/BRCA2 deficiency. In this review, we summarize a decade of PARP inhibitor clinical development, a work that has resulted in the registration of several PARP inhibitors in breast (olaparib and talazoparib) and ovarian cancer (olaparib, niraparib and rucaparib, either alone or following platinum chemotherapy as maintenance therapy). Over the past 10 years, our knowledge on the mechanism of action of PARP inhibitor as well as how tumours become resistant has been extended, and we summarise this work here. We also discuss opportunities for expanding the precision medicine approach with PARP inhibitors, identifying a wider population who could benefit from this drug class. This includes developing and validating better predictive biomarkers for patient stratification, mainly based on homologous recombination defects beyond BRCA1/BRCA2 mutations, identifying DNA repair deficient tumours in other cancer types such as prostate or pancreatic cancer, or by designing combination therapies with PARP inhibitors..
Evans, R.
Flores-Borja, F.
Nassiri, S.
Miranda, E.
Lawler, K.
Grigoriadis, A.
Monypenny, J.
Gillet, C.
Owen, J.
Gordon, P.
Male, V.
Cheung, A.
Noor, F.
Barber, P.
Marlow, R.
Francesch-Domenech, E.
Fruhwirth, G.
Squadrito, M.
Vojnovic, B.
Tutt, A.
Festy, F.
De Palma, M.
Ng, T.
(2019). Integrin-Mediated Macrophage Adhesion Promotes Lymphovascular Dissemination in Breast Cancer. Cell rep,
Vol.27
(7),
pp. 1967-1978.e4.
show abstract
full text
Lymphatic vasculature is crucial for metastasis in triple-negative breast cancer (TNBC); however, cellular and molecular drivers controlling lymphovascular metastasis are poorly understood. We define a macrophage-dependent signaling cascade that facilitates metastasis through lymphovascular remodeling. TNBC cells instigate mRNA changes in macrophages, resulting in β4 integrin-dependent adhesion to the lymphovasculature. β4 integrin retains macrophages proximal to lymphatic endothelial cells (LECs), where release of TGF-β1 drives LEC contraction via RhoA activation. Macrophages promote gross architectural changes to lymphovasculature by increasing dilation, hyperpermeability, and disorganization. TGF-β1 drives β4 integrin clustering at the macrophage plasma membrane, further promoting macrophage adhesion and demonstrating the dual functionality of TGF-β1 signaling in this context. β4 integrin-expressing macrophages were identified in human breast tumors, and a combination of vascular-remodeling macrophage gene signature and TGF-β signaling scores correlates with metastasis. We postulate that future clinical strategies for patients with TNBC should target crosstalk between β4 integrin and TGF-β1..
Fitzpatrick, A.
Tutt, A.
(2019). Controversial issues in the neoadjuvant treatment of triple-negative breast cancer. Ther adv med oncol,
Vol.11,
p. 1758835919882581.
show abstract
full text
Triple-negative breast cancer (TNBC), as a collective group of heterogenous tumours, displays the highest rate of distant recurrence and lowest survival from metastatic disease across breast cancer subtypes. However, a subset of TNBC display impressive primary tumour response to neoadjuvant chemotherapy, translating to reduction in future relapse and increased overall survival. Maximizing early treatment response is crucial to improving the outlook in this subtype. Numerous systemic therapy strategies are being assessed in the neoadjuvant setting and the current paradigm of generic chemotherapy components in regimens for high-risk breast cancers, regardless of biological subtype, is changing. Therapeutic approaches with evidence of benefit include platinum drugs, polyadenosine diphosphate ribose polymerase (PARP) inhibitors, immunotherapy and second adjuvant therapy for those not achieving pathological complete response. Importantly, molecular testing can identify subgroups within TNBC, such as deoxyribonucleic acid (DNA) homologous recombination repair deficiency, lymphocyte-predominant tumours, and TNBC type 4 molecular subtypes. Clinical trials that address the interaction between these biomarkers and treatment approaches are a priority, to identify subgroups benefiting from additional therapy..
Naidoo, K.
Wai, P.T.
Maguire, S.L.
Daley, F.
Haider, S.
Kriplani, D.
Campbell, J.
Mirza, H.
Grigoriadis, A.
Tutt, A.
Moseley, P.M.
Abdel-Fatah, T.M.
Chan, S.Y.
Madhusudan, S.
Rhaka, E.A.
Ellis, I.O.
Lord, C.J.
Yuan, Y.
Green, A.R.
Natrajan, R.
(2018). Evaluation of CDK12 Protein Expression as a Potential Novel Biomarker for DNA Damage Response-Targeted Therapies in Breast Cancer. Mol cancer ther,
Vol.17
(1),
pp. 306-315.
show abstract
full text
Disruption of Cyclin-Dependent Kinase 12 (CDK12) is known to lead to defects in DNA repair and sensitivity to platinum salts and PARP1/2 inhibitors. However, CDK12 has also been proposed as an oncogene in breast cancer. We therefore aimed to assess the frequency and distribution of CDK12 protein expression by IHC in independent cohorts of breast cancer and correlate this with outcome and genomic status. We found that 21% of primary unselected breast cancers were CDK12 high, and 10.5% were absent, by IHC. CDK12 positivity correlated with HER2 positivity but was not an independent predictor of breast cancer-specific survival taking HER2 status into account; however, absent CDK12 protein expression significantly correlated with a triple-negative phenotype. Interestingly, CDK12 protein absence was associated with reduced expression of a number of DDR proteins including ATR, Ku70/Ku80, PARP1, DNA-PK, and γH2AX, suggesting a novel mechanism of CDK12-associated DDR dysregulation in breast cancer. Our data suggest that diagnostic IHC quantification of CDK12 in breast cancer is feasible, with CDK12 absence possibly signifying defective DDR function. This may have important therapeutic implications, particularly for triple-negative breast cancers. Mol Cancer Ther; 17(1); 306-15. ©2017 AACR..
Tutt, A.
(2018). Inhibited, trapped or adducted: the optimal selective synthetic lethal mix for BRCAness. Ann oncol,
Vol.29
(1),
pp. 18-21.
full text
Grigoriadis, A.
Gazinska, P.
Pai, T.
Irhsad, S.
Wu, Y.
Millis, R.
Naidoo, K.
Owen, J.
Gillett, C.E.
Tutt, A.
Coolen, A.C.
Pinder, S.E.
(2018). Histological scoring of immune and stromal features in breast and axillary lymph nodes is prognostic for distant metastasis in lymph node-positive breast cancers. J pathol clin res,
Vol.4
(1),
pp. 39-54.
show abstract
full text
The prognostic importance of lymph node (LN) status and tumour-infiltrating lymphocytes (TILs), is well established, particularly TILs in triple negative breast cancers (TNBCs). So far, few studies have interrogated changes in involved and uninvolved LNs and evaluated if their morphological patterns add valuable information for the prediction of disease progression in breast cancer. In a cohort of 309 patients enriched for TNBCs (170/309), we histologically characterised immune and stromal features in primary tumours and associated involved and uninvolved axillary LNs on routine haematoxylin and eosin stained sections. Of the 309 patients, 143 had LN-positive disease. Twenty-five histopathological features were assessed, including the degree of TIL presence, quantitative and qualitative assessment of germinal centres (GCs) and sinus histiocytosis. Multivariate and cross-validated proportional hazard regression analyses were used to identify optimal covariate sets for prediction of distant metastasis-free survival (DMFS). The degree of intratumoural and peritumoural immune infiltrate was associated with architectural changes in both uninvolved and involved LNs. By including clinicopathological characteristics as well as tumour and LN histopathological features in L2-regularised proportional hazard models, the prediction of 5-year DMFS was improved by 3-15% over the baseline in all cancers and in TNBCs. In LN-positive cancers, the combination of Salgado's classification, lymphocytic lobulitis, size and number of GCs in the uninvolved LNs and location of GCs in the involved LNs carried significant prognostic information. From these features, a multivariate cross-validation-stable risk signature was constructed, which identified low-risk groups within both LN-positive breast cancers and the LN-positive TNBCs group with a 10-year DMFS probability of 78 and 87%, respectively. This study illustrates that, by incorporating histopathological patterns of involved and uninvolved LNs combined with primary tumour immune and stromal features, the prediction of developing distant metastasis in LN-positive breast cancers can be estimated more accurately..
Rodrigues, N.V.
Correia, D.V.
Mensurado, S.
Nóbrega-Pereira, S.
deBarros, A.
Kyle-Cezar, F.
Tutt, A.
Hayday, A.C.
Norell, H.
Silva-Santos, B.
Dias, S.
(2018). Low-Density Lipoprotein Uptake Inhibits the Activation and Antitumor Functions of Human Vγ9Vδ2 T Cells. Cancer immunology research,
Vol.6
(4),
pp. 448-457.
show abstract
Abstract
Vγ9Vδ2 T cells, the main subset of γδ T lymphocytes in human peripheral blood, are endowed with antitumor functions such as cytotoxicity and IFNγ production. These functions are triggered upon T-cell receptor–dependent activation by non-peptidic prenyl pyrophosphates (“phosphoantigens”) that are selective agonists of Vγ9Vδ2 T cells, and which have been evaluated in clinical studies. Because phosphoantigens have shown interindividual variation in Vγ9Vδ2 T-cell activities, we asked whether metabolic resources, namely lipids such as cholesterol, could affect phosphoantigen-mediated Vγ9Vδ2 T-cell activation and function. We show here that Vγ9Vδ2 T cells express the LDL receptor upon activation and take up LDL cholesterol. Resulting changes, such as decreased mitochondrial mass and reduced ATP production, correlate with downregulation of Vγ9Vδ2 T-cell activation and functionality. In particular, the expression of IFNγ, NKG2D, and DNAM-1 were reduced upon LDL cholesterol treatment of phosphoantigen-expanded Vγ9Vδ2 T cells. As a result, their capacity to target breast cancer cells was compromised both in vitro and in an in vivo xenograft mouse model. Thus, this study describes the role of LDL cholesterol as an inhibitor of the antitumor functions of phosphoantigen-activated Vγ9Vδ2 T cells. Our observations have implications for therapeutic applications dependent on Vγ9Vδ2 T cells. Cancer Immunol Res; 6(4); 448–57. ©2018 AACR..
Muliaditan, T.
Opzoomer, J.W.
Caron, J.
Okesola, M.
Kosti, P.
Lall, S.
Van Hemelrijck, M.
Dazzi, F.
Tutt, A.
Grigoriadis, A.
Gillett, C.E.
Madden, S.F.
Burchell, J.M.
Kordasti, S.
Diebold, S.S.
Spicer, J.F.
Arnold, J.N.
(2018). Repurposing Tin Mesoporphyrin as an Immune Checkpoint Inhibitor Shows Therapeutic Efficacy in Preclinical Models of Cancer. Clinical cancer research,
Vol.24
(7),
pp. 1617-1628.
Bajrami, I.
Marlow, R.
van de Ven, M.
Brough, R.
Pemberton, H.N.
Frankum, J.
Song, F.
Rafiq, R.
Konde, A.
Krastev, D.B.
Menon, M.
Campbell, J.
Gulati, A.
Kumar, R.
Pettitt, S.J.
Gurden, M.D.
Cardenosa, M.L.
Chong, I.
Gazinska, P.
Wallberg, F.
Sawyer, E.J.
Martin, L.-.
Dowsett, M.
Linardopoulos, S.
Natrajan, R.
Ryan, C.J.
Derksen, P.W.
Jonkers, J.
Tutt, A.N.
Ashworth, A.
Lord, C.J.
(2018). E-Cadherin/ROS1 Inhibitor Synthetic Lethality in Breast Cancer. Cancer discov,
Vol.8
(4),
pp. 498-515.
show abstract
full text
The cell adhesion glycoprotein E-cadherin (CDH1) is commonly inactivated in breast tumors. Precision medicine approaches that exploit this characteristic are not available. Using perturbation screens in breast tumor cells with CRISPR/Cas9-engineered CDH1 mutations, we identified synthetic lethality between E-cadherin deficiency and inhibition of the tyrosine kinase ROS1. Data from large-scale genetic screens in molecularly diverse breast tumor cell lines established that the E-cadherin/ROS1 synthetic lethality was not only robust in the face of considerable molecular heterogeneity but was also elicited with clinical ROS1 inhibitors, including foretinib and crizotinib. ROS1 inhibitors induced mitotic abnormalities and multinucleation in E-cadherin-defective cells, phenotypes associated with a defect in cytokinesis and aberrant p120 catenin phosphorylation and localization. In vivo, ROS1 inhibitors produced profound antitumor effects in multiple models of E-cadherin-defective breast cancer. These data therefore provide the preclinical rationale for assessing ROS1 inhibitors, such as the licensed drug crizotinib, in appropriately stratified patients.Significance: E-cadherin defects are common in breast cancer but are currently not targeted with a precision medicine approach. Our preclinical data indicate that licensed ROS1 inhibitors, including crizotinib, should be repurposed to target E-cadherin-defective breast cancers, thus providing the rationale for the assessment of these agents in molecularly stratified phase II clinical trials. Cancer Discov; 8(4); 498-515. ©2018 AACR.This article is highlighted in the In This Issue feature, p. 371..
Tutt, A.
Tovey, H.
Cheang, M.C.
Kernaghan, S.
Kilburn, L.
Gazinska, P.
Owen, J.
Abraham, J.
Barrett, S.
Barrett-Lee, P.
Brown, R.
Chan, S.
Dowsett, M.
Flanagan, J.M.
Fox, L.
Grigoriadis, A.
Gutin, A.
Harper-Wynne, C.
Hatton, M.Q.
Hoadley, K.A.
Parikh, J.
Parker, P.
Perou, C.M.
Roylance, R.
Shah, V.
Shaw, A.
Smith, I.E.
Timms, K.M.
Wardley, A.M.
Wilson, G.
Gillett, C.
Lanchbury, J.S.
Ashworth, A.
Rahman, N.
Harries, M.
Ellis, P.
Pinder, S.E.
Bliss, J.M.
(2018). Carboplatin in BRCA1/2-mutated and triple-negative breast cancer BRCAness subgroups: the TNT Trial. Nat med,
Vol.24
(5),
pp. 628-637.
show abstract
full text
Germline mutations in BRCA1/2 predispose individuals to breast cancer (termed germline-mutated BRCA1/2 breast cancer, gBRCA-BC) by impairing homologous recombination (HR) and causing genomic instability. HR also repairs DNA lesions caused by platinum agents and PARP inhibitors. Triple-negative breast cancers (TNBCs) harbor subpopulations with BRCA1/2 mutations, hypothesized to be especially platinum-sensitive. Cancers in putative 'BRCAness' subgroups-tumors with BRCA1 methylation; low levels of BRCA1 mRNA (BRCA1 mRNA-low); or mutational signatures for HR deficiency and those with basal phenotypes-may also be sensitive to platinum. We assessed the efficacy of carboplatin and another mechanistically distinct therapy, docetaxel, in a phase 3 trial in subjects with unselected advanced TNBC. A prespecified protocol enabled biomarker-treatment interaction analyses in gBRCA-BC and BRCAness subgroups. The primary endpoint was objective response rate (ORR). In the unselected population (376 subjects; 188 carboplatin, 188 docetaxel), carboplatin was not more active than docetaxel (ORR, 31.4% versus 34.0%, respectively; P = 0.66). In contrast, in subjects with gBRCA-BC, carboplatin had double the ORR of docetaxel (68% versus 33%, respectively; biomarker, treatment interaction P = 0.01). Such benefit was not observed for subjects with BRCA1 methylation, BRCA1 mRNA-low tumors or a high score in a Myriad HRD assay. Significant interaction between treatment and the basal-like subtype was driven by high docetaxel response in the nonbasal subgroup. We conclude that patients with advanced TNBC benefit from characterization of BRCA1/2 mutations, but not BRCA1 methylation or Myriad HRD analyses, to inform choices on platinum-based chemotherapy. Additionally, gene expression analysis of basal-like cancers may also influence treatment selection..
Patel, N.
Weekes, D.
Drosopoulos, K.
Gazinska, P.
Noel, E.
Rashid, M.
Mirza, H.
Quist, J.
Brasó-Maristany, F.
Mathew, S.
Ferro, R.
Pereira, A.M.
Prince, C.
Noor, F.
Francesch-Domenech, E.
Marlow, R.
de Rinaldis, E.
Grigoriadis, A.
Linardopoulos, S.
Marra, P.
Tutt, A.N.
(2018). Integrated genomics and functional validation identifies malignant cell specific dependencies in triple negative breast cancer. Nat commun,
Vol.9
(1),
p. 1044.
show abstract
full text
Triple negative breast cancers (TNBCs) lack recurrent targetable driver mutations but demonstrate frequent copy number aberrations (CNAs). Here, we describe an integrative genomic and RNAi-based approach that identifies and validates gene addictions in TNBCs. CNAs and gene expression alterations are integrated and genes scored for pre-specified target features revealing 130 candidate genes. We test functional dependence on each of these genes using RNAi in breast cancer and non-malignant cells, validating malignant cell selective dependence upon 37 of 130 genes. Further analysis reveals a cluster of 13 TNBC addiction genes frequently co-upregulated that includes genes regulating cell cycle checkpoints, DNA damage response, and malignant cell selective mitotic genes. We validate the mechanism of addiction to a potential drug target: the mitotic kinesin family member C1 (KIFC1/HSET), essential for successful bipolar division of centrosome-amplified malignant cells and develop a potential selection biomarker to identify patients with tumors exhibiting centrosome amplification..
Irshad, S.
Flores-Borja, F.
Lawler, K.
Monypenny, J.
Evans, R.
Male, V.
Gordon, P.
Cheung, A.
Gazinska, P.
Noor, F.
Wong, F.
Grigoriadis, A.
Fruhwirth, G.O.
Barber, P.R.
Woodman, N.
Patel, D.
Rodriguez-Justo, M.
Owen, J.
Martin, S.G.
Pinder, S.E.
Gillett, C.E.
Poland, S.P.
Ameer-Beg, S.
McCaughan, F.
Carlin, L.M.
Hasan, U.
Withers, D.R.
Lane, P.
Vojnovic, B.
Quezada, S.A.
Ellis, P.
Tutt, A.N.
Ng, T.
(2017). RORγt + Innate Lymphoid Cells Promote Lymph Node Metastasis of Breast Cancers. Cancer research,
Vol.77
(5),
pp. 1083-1096.
full text
Curigliano, G.
Burstein, H.J.
Winer, E.P.
Gnant, M.
Dubsky, P.
Loibl, S.
Colleoni, M.
Regan, M.M.
Piccart-Gebhart, M.
Senn, H.-.
Thürlimann, B.
St. Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2017,
André, F.
Baselga, J.
Bergh, J.
Bonnefoi, H.
Brucker, S.Y.
Cardoso, F.
Carey, L.
Ciruelos, E.
Cuzick, J.
Denkert, C.
Di Leo, A.
Ejlertsen, B.
Francis, P.
Galimberti, V.
Garber, J.
Gulluoglu, B.
Goodwin, P.
Harbeck, N.
Hayes, D.F.
Huang, C.-.
Huober, J.
Hussein, K.
Jassem, J.
Jiang, Z.
Karlsson, P.
Morrow, M.
Orecchia, R.
Osborne, K.C.
Pagani, O.
Partridge, A.H.
Pritchard, K.
Ro, J.
Rutgers, E.J.
Sedlmayer, F.
Semiglazov, V.
Shao, Z.
Smith, I.
Toi, M.
Tutt, A.
Viale, G.
Watanabe, T.
Whelan, T.J.
Xu, B.
(2017). De-escalating and escalating treatments for early-stage breast cancer: the St Gallen International Expert Consensus Conference on the Primary Therapy of Early Breast Cancer 2017. Ann oncol,
Vol.28
(8),
pp. 1700-1712.
show abstract
full text
The 15th St. Gallen International Breast Cancer Conference 2017 in Vienna, Austria reviewed substantial new evidence on loco-regional and systemic therapies for early breast cancer. Treatments were assessed in light of their intensity, duration and side-effects, seeking where appropriate to escalate or de-escalate therapies based on likely benefits as predicted by tumor stage and tumor biology. The Panel favored several interventions that may reduce surgical morbidity, including acceptance of 2 mm margins for DCIS, the resection of residual cancer (but not baseline extent of cancer) in women undergoing neoadjuvant therapy, acceptance of sentinel node biopsy following neoadjuvant treatment of many patients, and the preference for neoadjuvant therapy in HER2 positive and triple-negative, stage II and III breast cancer. The Panel favored escalating radiation therapy with regional nodal irradiation in high-risk patients, while encouraging omission of boost in low-risk patients. The Panel endorsed gene expression signatures that permit avoidance of chemotherapy in many patients with ER positive breast cancer. For women with higher risk tumors, the Panel escalated recommendations for adjuvant endocrine treatment to include ovarian suppression in premenopausal women, and extended therapy for postmenopausal women. However, low-risk patients can avoid these treatments. Finally, the Panel recommended bisphosphonate use in postmenopausal women to prevent breast cancer recurrence. The Panel recognized that recommendations are not intended for all patients, but rather to address the clinical needs of the majority of common presentations. Individualization of adjuvant therapy means adjusting to the tumor characteristics, patient comorbidities and preferences, and managing constraints of treatment cost and access that may affect care in both the developed and developing world..
Davies, H.
Glodzik, D.
Morganella, S.
Yates, L.R.
Staaf, J.
Zou, X.
Ramakrishna, M.
Martin, S.
Boyault, S.
Sieuwerts, A.M.
Simpson, P.T.
King, T.A.
Raine, K.
Eyfjord, J.E.
Kong, G.
Borg, Å.
Birney, E.
Stunnenberg, H.G.
van de Vijver, M.J.
Børresen-Dale, A.-.
Martens, J.W.
Span, P.N.
Lakhani, S.R.
Vincent-Salomon, A.
Sotiriou, C.
Tutt, A.
Thompson, A.M.
Van Laere, S.
Richardson, A.L.
Viari, A.
Campbell, P.J.
Stratton, M.R.
Nik-Zainal, S.
(2017). HRDetect is a predictor of BRCA1 and BRCA2 deficiency based on mutational signatures. Nat med,
Vol.23
(4),
pp. 517-525.
show abstract
full text
Approximately 1-5% of breast cancers are attributed to inherited mutations in BRCA1 or BRCA2 and are selectively sensitive to poly(ADP-ribose) polymerase (PARP) inhibitors. In other cancer types, germline and/or somatic mutations in BRCA1 and/or BRCA2 (BRCA1/BRCA2) also confer selective sensitivity to PARP inhibitors. Thus, assays to detect BRCA1/BRCA2-deficient tumors have been sought. Recently, somatic substitution, insertion/deletion and rearrangement patterns, or 'mutational signatures', were associated with BRCA1/BRCA2 dysfunction. Herein we used a lasso logistic regression model to identify six distinguishing mutational signatures predictive of BRCA1/BRCA2 deficiency. A weighted model called HRDetect was developed to accurately detect BRCA1/BRCA2-deficient samples. HRDetect identifies BRCA1/BRCA2-deficient tumors with 98.7% sensitivity (area under the curve (AUC) = 0.98). Application of this model in a cohort of 560 individuals with breast cancer, of whom 22 were known to carry a germline BRCA1 or BRCA2 mutation, allowed us to identify an additional 22 tumors with somatic loss of BRCA1 or BRCA2 and 47 tumors with functional BRCA1/BRCA2 deficiency where no mutation was detected. We validated HRDetect on independent cohorts of breast, ovarian and pancreatic cancers and demonstrated its efficacy in alternative sequencing strategies. Integrating all of the classes of mutational signatures thus reveals a larger proportion of individuals with breast cancer harboring BRCA1/BRCA2 deficiency (up to 22%) than hitherto appreciated (∼1-5%) who could have selective therapeutic sensitivity to PARP inhibition..
Gracio, F.
Burford, B.
Gazinska, P.
Mera, A.
Mohd Noor, A.
Marra, P.
Gillett, C.
Grigoriadis, A.
Pinder, S.
Tutt, A.
de Rinaldis, E.
(2017). Splicing imbalances in basal-like breast cancer underpin perturbation of cell surface and oncogenic pathways and are associated with patients' survival. Sci rep,
Vol.7,
p. 40177.
show abstract
full text
Despite advancements in the use of transcriptional information to understand and classify breast cancers, the contribution of splicing to the establishment and progression of these tumours has only recently starting to emerge. Our work explores this lesser known landscape, with special focus on the basal-like breast cancer subtype where limited therapeutic opportunities and no prognostic biomarkers are currently available. Using ExonArray analysis of 176 breast cancers and 9 normal breast tissues we demonstrate that splicing levels significantly contribute to the diversity of breast cancer molecular subtypes and explain much of the differences compared with normal tissues. We identified pathways specifically affected by splicing imbalances whose perturbation would be hidden from a conventional gene-centric analysis of gene expression. We found that a large fraction of them involve cell-to-cell communication, extracellular matrix and transport, as well as oncogenic and immune-related pathways transduced by plasma membrane receptors. We identified 247 genes in which splicing imbalances are associated with clinical patients' outcome, whilst no association was detectable at the gene expression level. These include the signaling gene TGFBR1, the proto-oncogene MYB as well as many immune-related genes such as CCR7 and FCRL3, reinforcing evidence for a role of immune components in influencing breast cancer patients' prognosis..
Coles, C.E.
Griffin, C.L.
Kirby, A.M.
Titley, J.
Agrawal, R.K.
Alhasso, A.
Bhattacharya, I.S.
Brunt, A.M.
Ciurlionis, L.
Chan, C.
Donovan, E.M.
Emson, M.A.
Harnett, A.N.
Haviland, J.S.
Hopwood, P.
Jefford, M.L.
Kaggwa, R.
Sawyer, E.J.
Syndikus, I.
Tsang, Y.M.
Wheatley, D.A.
Wilcox, M.
Yarnold, J.R.
Bliss, J.M.
IMPORT Trialists,
(2017). Partial-breast radiotherapy after breast conservation surgery for patients with early breast cancer (UK IMPORT LOW trial): 5-year results from a multicentre, randomised, controlled, phase 3, non-inferiority trial. Lancet,
Vol.390
(10099),
pp. 1048-1060.
show abstract
full text
BACKGROUND: Local cancer relapse risk after breast conservation surgery followed by radiotherapy has fallen sharply in many countries, and is influenced by patient age and clinicopathological factors. We hypothesise that partial-breast radiotherapy restricted to the vicinity of the original tumour in women at lower than average risk of local relapse will improve the balance of beneficial versus adverse effects compared with whole-breast radiotherapy. METHODS: IMPORT LOW is a multicentre, randomised, controlled, phase 3, non-inferiority trial done in 30 radiotherapy centres in the UK. Women aged 50 years or older who had undergone breast-conserving surgery for unifocal invasive ductal adenocarcinoma of grade 1-3, with a tumour size of 3 cm or less (pT1-2), none to three positive axillary nodes (pN0-1), and minimum microscopic margins of non-cancerous tissue of 2 mm or more, were recruited. Patients were randomly assigned (1:1:1) to receive 40 Gy whole-breast radiotherapy (control), 36 Gy whole-breast radiotherapy and 40 Gy to the partial breast (reduced-dose group), or 40 Gy to the partial breast only (partial-breast group) in 15 daily treatment fractions. Computer-generated random permuted blocks (mixed sizes of six and nine) were used to assign patients to groups, stratifying patients by radiotherapy treatment centre. Patients and clinicians were not masked to treatment allocation. Field-in-field intensity-modulated radiotherapy was delivered using standard tangential beams that were simply reduced in length for the partial-breast group. The primary endpoint was ipsilateral local relapse (80% power to exclude a 2·5% increase [non-inferiority margin] at 5 years for each experimental group; non-inferiority was shown if the upper limit of the two-sided 95% CI for the local relapse hazard ratio [HR] was less than 2·03), analysed by intention to treat. Safety analyses were done in all patients for whom data was available (ie, a modified intention-to-treat population). This study is registered in the ISRCTN registry, number ISRCTN12852634. FINDINGS: Between May 3, 2007, and Oct 5, 2010, 2018 women were recruited. Two women withdrew consent for use of their data in the analysis. 674 patients were analysed in the whole-breast radiotherapy (control) group, 673 in the reduced-dose group, and 669 in the partial-breast group. Median follow-up was 72·2 months (IQR 61·7-83·2), and 5-year estimates of local relapse cumulative incidence were 1·1% (95% CI 0·5-2·3) of patients in the control group, 0·2% (0·02-1·2) in the reduced-dose group, and 0·5% (0·2-1·4) in the partial-breast group. Estimated 5-year absolute differences in local relapse compared with the control group were -0·73% (-0·99 to 0·22) for the reduced-dose and -0·38% (-0·84 to 0·90) for the partial-breast groups. Non-inferiority can be claimed for both reduced-dose and partial-breast radiotherapy, and was confirmed by the test against the critical HR being more than 2·03 (p=0·003 for the reduced-dose group and p=0·016 for the partial-breast group, compared with the whole-breast radiotherapy group). Photographic, patient, and clinical assessments recorded similar adverse effects after reduced-dose or partial-breast radiotherapy, including two patient domains achieving statistically significantly lower adverse effects (change in breast appearance [p=0·007 for partial-breast] and breast harder or firmer [p=0·002 for reduced-dose and p<0·0001 for partial-breast]) compared with whole-breast radiotherapy. INTERPRETATION: We showed non-inferiority of partial-breast and reduced-dose radiotherapy compared with the standard whole-breast radiotherapy in terms of local relapse in a cohort of patients with early breast cancer, and equivalent or fewer late normal-tissue adverse effects were seen. This simple radiotherapy technique is implementable in radiotherapy centres worldwide. FUNDING: Cancer Research UK..
Koo, C.-.
Giacomini, C.
Reyes-Corral, M.
Olmos, Y.
Tavares, I.A.
Marson, C.M.
Linardopoulos, S.
Tutt, A.N.
Morris, J.D.
(2017). Targeting TAO Kinases Using a New Inhibitor Compound Delays Mitosis and Induces Mitotic Cell Death in Centrosome Amplified Breast Cancer Cells. Mol cancer ther,
Vol.16
(11),
pp. 2410-2421.
show abstract
full text
Thousand-and-one amino acid kinases (TAOK) 1 and 2 are activated catalytically during mitosis and can contribute to mitotic cell rounding and spindle positioning. Here, we characterize a compound that inhibits TAOK1 and TAOK2 activity with IC50 values of 11 to 15 nmol/L, is ATP-competitive, and targets these kinases selectively. TAOK inhibition or depletion in centrosome-amplified SKBR3 or BT549 breast cancer cell models increases the mitotic population, the percentages of mitotic cells displaying amplified centrosomes and multipolar spindles, induces cell death, and inhibits cell growth. In contrast, nontumorigenic and dividing bipolar MCF-10A breast cells appear less dependent on TAOK activity and can complete mitosis and proliferate in the presence of the TAOK inhibitor. We demonstrate that TAOK1 and TAOK2 localize to the cytoplasm and centrosomes respectively during mitosis. Live cell imaging shows that the TAOK inhibitor prolongs the duration of mitosis in SKBR3 cells, increases mitotic cell death, and reduces the percentages of cells exiting mitosis, whereas MCF-10A cells continue to divide and proliferate. Over 80% of breast cancer tissues display supernumerary centrosomes, and tumor cells frequently cluster extra centrosomes to avoid multipolar mitoses and associated cell death. Consequently, drugs that stimulate centrosome declustering and induce multipolarity are likely to target dividing centrosome-amplified cancer cells preferentially, while sparing normal bipolar cells. Our results demonstrate that TAOK inhibition can enhance centrosome declustering and mitotic catastrophe in cancer cells, and these proteins may therefore offer novel therapeutic targets suitable for drug inhibition and the potential treatment of breast cancers, where supernumerary centrosomes occur. Mol Cancer Ther; 16(11); 2410-21. ©2017 AACR..
Lawler, K.
Papouli, E.
Naceur-Lombardelli, C.
Mera, A.
Ougham, K.
Tutt, A.
Kimbung, S.
Hedenfalk, I.
Zhan, J.
Zhang, H.
Buus, R.
Dowsett, M.
Ng, T.
Pinder, S.E.
Parker, P.
Holmberg, L.
Gillett, C.E.
Grigoriadis, A.
Purushotham, A.
(2017). Gene expression modules in primary breast cancers as risk factors for organotropic patterns of first metastatic spread: a case control study. Breast cancer res,
Vol.19
(1),
p. 113.
show abstract
full text
BACKGROUND: Metastases from primary breast cancers can involve single or multiple organs at metastatic disease diagnosis. Molecular risk factors for particular patterns of metastastic spread in a clinical population are limited. METHODS: A case-control design including 1357 primary breast cancers was used to study three distinct clinical patterns of metastasis, which occur within the first six months of metastatic disease: bone and visceral metasynchronous spread, bone-only, and visceral-only metastasis. Whole-genome expression profiles were obtained using whole genome (WG)-DASL assays from formalin-fixed paraffin-embedded (FFPE) samples. A systematic protocol was developed for handling FFPE samples together with stringent data quality controls to identify robust expression profiling data. A panel of published and novel gene sets were tested for association with these specific patterns of metastatic spread and odds ratios (ORs) were calculated. RESULTS: Metasynchronous metastasis to bone and viscera was found in all intrinsic breast cancer subtypes, while immunohistochemically (IHC)-defined receptor status and specific IntClust subgroups were risk factors for visceral-only or bone-only first metastases. Among gene modules, those related to proliferation increased the risk of metasynchronous metastasis (OR (95% CI) = 2.3 (1.1-4.8)) and visceral-only first metastasis (OR (95% CI) = 2.5 (1.2-5.1)) but not bone-only metastasis (OR (95% CI) = 0.97 (0.56-1.7)). A 21-gene module (BV) was identified in estrogen-receptor-positive breast cancers with metasynchronous metastasis to bone and viscera (area under the curve = 0.77), and its expression increased the risk of bone and visceral metasynchronous spread in this population. BV was further orthogonally validated with NanoString nCounter in primary breast cancers, and was reproducible in their matched lymph nodes metastases and an external cohort. CONCLUSION: This case-control study of WG-DASL global expression profiles from FFPE tumour samples, after careful quality control and RNA selection, revealed that gene modules in the primary tumour have differing risks for clinical patterns of metasynchronous first metastases. Moreover, a novel gene module was identified as a putative risk factor for metasynchronous bone and visceral first metastatic spread, with potential implications for disease monitoring and treatment planning..
Yates, L.R.
Knappskog, S.
Wedge, D.
Farmery, J.H.
Gonzalez, S.
Martincorena, I.
Alexandrov, L.B.
Van Loo, P.
Haugland, H.K.
Lilleng, P.K.
Gundem, G.
Gerstung, M.
Pappaemmanuil, E.
Gazinska, P.
Bhosle, S.G.
Jones, D.
Raine, K.
Mudie, L.
Latimer, C.
Sawyer, E.
Desmedt, C.
Sotiriou, C.
Stratton, M.R.
Sieuwerts, A.M.
Lynch, A.G.
Martens, J.W.
Richardson, A.L.
Tutt, A.
Lonning, P.E.
Campbell, P.J.
(2017). Genomic Evolution of Breast Cancer Metastasis and Relapse. Cancer cell,
Vol.32
(2),
pp. 169-+.
full text
Eisenblaetter, M.
Flores-Borja, F.
Lee, J.J.
Wefers, C.
Smith, H.
Hueting, R.
Cooper, M.S.
Blower, P.J.
Patel, D.
Rodriguez-Justo, M.
Milewicz, H.
Vogl, T.
Roth, J.
Tutt, A.
Schaeffter, T.
Ng, T.
(2017). Visualization of Tumor-Immune Interaction - Target-Specific Imaging of S100A8/A9 Reveals Pre-Metastatic Niche Establishment. Theranostics,
Vol.7
(9),
pp. 2392-2401.
full text
Denkert, C.
Liedtke, C.
Tutt, A.
von Minckwitz, G.
(2017). Molecular alterations in triple-negative breast cancer-the road to new treatment strategies. Lancet,
Vol.389
(10087),
pp. 2430-2442.
show abstract
full text
Triple-negative breast cancer is a heterogeneous disease and specific therapies have not been available for a long time. Therefore, conventional chemotherapy is still considered the clinical state of the art. Different subgroups of triple-negative breast cancer have been identified on the basis of protein expression, mRNA signatures, and genomic alterations. Important elements of triple-negative breast cancer biology include high proliferative activity, an increased immunological infiltrate, a basal-like and a mesenchymal phenotype, and deficiency in homologous recombination, which is in part associated with loss of BRCA1 or BRCA2 function. A minority of triple-negative tumours express luminal markers, such as androgen receptors, and have a lower proliferative activity. These biological subgroups are overlapping and currently cannot be combined into a unified model of triple-negative breast cancer biology. Nevertheless, the molecular analysis of this disease has identified potential options for targeted therapeutic intervention. This has led to promising clinical strategies, including modified chemotherapy approaches targeting the DNA damage response, angiogenesis inhibitors, immune checkpoint inhibitors, or even anti-androgens, all of which are being evaluated in phase 1-3 clinical studies. This Series paper focuses on the most relevant clinical questions, summarises the results of recent clinical trials, and gives an overview of ongoing studies and trial concepts that will lead to a more refined therapy for this tumour type..
Ju, Y.S.
Martincorena, I.
Gerstung, M.
Petljak, M.
Alexandrov, L.B.
Rahbari, R.
Wedge, D.C.
Davies, H.R.
Ramakrishna, M.
Fullam, A.
Martin, S.
Alder, C.
Patel, N.
Gamble, S.
O'Meara, S.
Giri, D.D.
Sauer, T.
Pinder, S.E.
Purdie, C.A.
Borg, Å.
Stunnenberg, H.
van de Vijver, M.
Tan, B.K.
Caldas, C.
Tutt, A.
Ueno, N.T.
van 't Veer, L.J.
Martens, J.W.
Sotiriou, C.
Knappskog, S.
Span, P.N.
Lakhani, S.R.
Eyfjörd, J.E.
Børresen-Dale, A.-.
Richardson, A.
Thompson, A.M.
Viari, A.
Hurles, M.E.
Nik-Zainal, S.
Campbell, P.J.
Stratton, M.R.
(2017). Somatic mutations reveal asymmetric cellular dynamics in the early human embryo. Nature,
Vol.543
(7647),
pp. 714-718.
show abstract
full text
Somatic cells acquire mutations throughout the course of an individual's life. Mutations occurring early in embryogenesis are often present in a substantial proportion of, but not all, cells in postnatal humans and thus have particular characteristics and effects. Depending on their location in the genome and the proportion of cells they are present in, these mosaic mutations can cause a wide range of genetic disease syndromes and predispose carriers to cancer. They have a high chance of being transmitted to offspring as de novo germline mutations and, in principle, can provide insights into early human embryonic cell lineages and their contributions to adult tissues. Although it is known that gross chromosomal abnormalities are remarkably common in early human embryos, our understanding of early embryonic somatic mutations is very limited. Here we use whole-genome sequences of normal blood from 241 adults to identify 163 early embryonic mutations. We estimate that approximately three base substitution mutations occur per cell per cell-doubling event in early human embryogenesis and these are mainly attributable to two known mutational signatures. We used the mutations to reconstruct developmental lineages of adult cells and demonstrate that the two daughter cells of many early embryonic cell-doubling events contribute asymmetrically to adult blood at an approximately 2:1 ratio. This study therefore provides insights into the mutation rates, mutational processes and developmental outcomes of cell dynamics that operate during early human embryogenesis..
Fittall, M.W.
Cheung, A.
Petranyi, G.M.
Rodriguez-Dominguez, D.
Bax, H.J.
Karagiannis, P.
Ilieva, K.M.
Tutt, A.
Karagiannis, S.N.
(2016). Exploring folate receptor a immunotherapy of breast carcinomas: Human monocytic cell-mediated killing triggered by IgG1 and IgE antibodies. Cancer immunology research,
Vol.4
(1).
Fittall, M.W.
Levi, D.
Clifford, A.
Shah, V.
Cheung, A.
Tutt, A.
Karagiannis, S.N.
(2016). The circulating memory B cell compartment of breast cancer patients is depleted in comparison with healthy volunteers. Cancer immunology research,
Vol.4
(1).
Marra, P.
Mathew, S.
Catchpole, S.
Facchetti, A.
Tutt, A.
(2016). Interferon gamma and post-translational modifications control the dynamics and plasma membrane association of IL15RA expressed by Triple Negative Breast Cancer (TNBC). Cancer immunology research,
Vol.4
(1).
Nik-Zainal, S.
Davies, H.
Staaf, J.
Ramakrishna, M.
Glodzik, D.
Zou, X.
Martincorena, I.
Alexandrov, L.B.
Martin, S.
Wedge, D.C.
Van Loo, P.
Ju, Y.S.
Smid, M.
Brinkman, A.B.
Morganella, S.
Aure, M.R.
Lingjærde, O.C.
Langerød, A.
Ringnér, M.
Ahn, S.-.
Boyault, S.
Brock, J.E.
Broeks, A.
Butler, A.
Desmedt, C.
Dirix, L.
Dronov, S.
Fatima, A.
Foekens, J.A.
Gerstung, M.
Hooijer, G.K.
Jang, S.J.
Jones, D.R.
Kim, H.-.
King, T.A.
Krishnamurthy, S.
Lee, H.J.
Lee, J.-.
Li, Y.
McLaren, S.
Menzies, A.
Mustonen, V.
O'Meara, S.
Pauporté, I.
Pivot, X.
Purdie, C.A.
Raine, K.
Ramakrishnan, K.
Rodríguez-González, F.G.
Romieu, G.
Sieuwerts, A.M.
Simpson, P.T.
Shepherd, R.
Stebbings, L.
Stefansson, O.A.
Teague, J.
Tommasi, S.
Treilleux, I.
Van den Eynden, G.G.
Vermeulen, P.
Vincent-Salomon, A.
Yates, L.
Caldas, C.
van't Veer, L.
Tutt, A.
Knappskog, S.
Tan, B.K.
Jonkers, J.
Borg, Å.
Ueno, N.T.
Sotiriou, C.
Viari, A.
Futreal, P.A.
Campbell, P.J.
Span, P.N.
Van Laere, S.
Lakhani, S.R.
Eyfjord, J.E.
Thompson, A.M.
Birney, E.
Stunnenberg, H.G.
van de Vijver, M.J.
Martens, J.W.
Børresen-Dale, A.-.
Richardson, A.L.
Kong, G.
Thomas, G.
Stratton, M.R.
(2016). Landscape of somatic mutations in 560 breast cancer whole-genome sequences. Nature,
Vol.534
(7605),
pp. 47-54.
show abstract
full text
We analysed whole-genome sequences of 560 breast cancers to advance understanding of the driver mutations conferring clonal advantage and the mutational processes generating somatic mutations. We found that 93 protein-coding cancer genes carried probable driver mutations. Some non-coding regions exhibited high mutation frequencies, but most have distinctive structural features probably causing elevated mutation rates and do not contain driver mutations. Mutational signature analysis was extended to genome rearrangements and revealed twelve base substitution and six rearrangement signatures. Three rearrangement signatures, characterized by tandem duplications or deletions, appear associated with defective homologous-recombination-based DNA repair: one with deficient BRCA1 function, another with deficient BRCA1 or BRCA2 function, the cause of the third is unknown. This analysis of all classes of somatic mutation across exons, introns and intergenic regions highlights the repertoire of cancer genes and mutational processes operating, and progresses towards a comprehensive account of the somatic genetic basis of breast cancer..
Watkins, J.
Tutt, A.
Grigoriadis, A.
(2016). Tandem duplications contribute to not one but two distinct phenotypes. Proc natl acad sci u s a,
Vol.113
(36),
pp. E5257-E5258.
full text
Cheung, A.
Bax, H.J.
Josephs, D.H.
Ilieva, K.M.
Pellizzari, G.
Opzoomer, J.
Bloomfield, J.
Fittall, M.
Grigoriadis, A.
Figini, M.
Canevari, S.
Spicer, J.F.
Tutt, A.N.
Karagiannis, S.N.
(2016). Targeting folate receptor alpha for cancer treatment. Oncotarget,
Vol.7
(32),
pp. 52553-52574.
show abstract
full text
Promising targeted treatments and immunotherapy strategies in oncology and advancements in our understanding of molecular pathways that underpin cancer development have reignited interest in the tumor-associated antigen Folate Receptor alpha (FRα). FRα is a glycosylphosphatidylinositol (GPI)-anchored membrane protein. Its overexpression in tumors such as ovarian, breast and lung cancers, low and restricted distribution in normal tissues, alongside emerging insights into tumor-promoting functions and association of expression with patient prognosis, together render FRα an attractive therapeutic target. In this review, we summarize the role of FRα in cancer development, we consider FRα as a potential diagnostic and prognostic tool, and we discuss different targeted treatment approaches with a specific focus on monoclonal antibodies. Renewed attention to FRα may point to novel individualized treatment approaches to improve the clinical management of patient groups that do not adequately benefit from current conventional therapies..
Brasó-Maristany, F.
Filosto, S.
Catchpole, S.
Marlow, R.
Quist, J.
Francesch-Domenech, E.
Plumb, D.A.
Zakka, L.
Gazinska, P.
Liccardi, G.
Meier, P.
Gris-Oliver, A.
Cheang, M.C.
Perdrix-Rosell, A.
Shafat, M.
Noël, E.
Patel, N.
McEachern, K.
Scaltriti, M.
Castel, P.
Noor, F.
Buus, R.
Mathew, S.
Watkins, J.
Serra, V.
Marra, P.
Grigoriadis, A.
Tutt, A.N.
(2016). PIM1 kinase regulates cell death, tumor growth and chemotherapy response in triple-negative breast cancer. Nat med,
Vol.22
(11),
pp. 1303-1313.
show abstract
full text
Triple-negative breast cancers (TNBCs) have poor prognosis and lack targeted therapies. Here we identified increased copy number and expression of the PIM1 proto-oncogene in genomic data sets of patients with TNBC. TNBC cells, but not nonmalignant mammary epithelial cells, were dependent on PIM1 for proliferation and protection from apoptosis. PIM1 knockdown reduced expression of the anti-apoptotic factor BCL2, and dynamic BH3 profiling of apoptotic priming revealed that PIM1 prevents mitochondrial-mediated apoptosis in TNBC cell lines. In TNBC tumors and their cellular models, PIM1 expression was associated with several transcriptional signatures involving the transcription factor MYC, and PIM1 depletion in TNBC cell lines decreased, in a MYC-dependent manner, cell population growth and expression of the MYC target gene MCL1. Treatment with the pan-PIM kinase inhibitor AZD1208 impaired the growth of both cell line and patient-derived xenografts and sensitized them to standard-of-care chemotherapy. This work identifies PIM1 as a malignant-cell-selective target in TNBC and the potential use of PIM1 inhibitors for sensitizing TNBC to chemotherapy-induced apoptotic cell death..
Flores-Borja, F.
Irshad, S.
Gordon, P.
Wong, F.
Sheriff, I.
Tutt, A.
Ng, T.
(2016). Crosstalk between Innate Lymphoid Cells and Other Immune Cells in the Tumor Microenvironment. J immunol res,
Vol.2016,
p. 7803091.
show abstract
full text
Our knowledge and understanding of the tumor microenvironment (TME) have been recently expanded with the recognition of the important role of innate lymphoid cells (ILC). Three different groups of ILC have been described based on their ability to produce cytokines that mediate the interactions between innate and adaptive immune cells in a variety of immune responses in infection, allergy, and autoimmunity. However, recent evidence from experimental models and clinical studies has demonstrated that ILC contribute to the mechanisms that generate suppressive or tolerant environments that allow tumor regression or progression. Defining the complex network of interactions and crosstalk of ILC with other immune cells and understanding the specific contributions of each type of ILC leading to tumor development will allow the manipulation of their function and will be important to develop new interventions and therapeutic strategies..
Akre, M.K.
Starrett, G.J.
Quist, J.S.
Temiz, N.A.
Carpenter, M.A.
Tutt, A.N.
Grigoriadis, A.
Harris, R.S.
(2016). Mutation Processes in 293-Based Clones Overexpressing the DNA Cytosine Deaminase APOBEC3B. Plos one,
Vol.11
(5),
p. e0155391.
show abstract
Molecular, cellular, and clinical studies have combined to demonstrate a contribution from the DNA cytosine deaminase APOBEC3B (A3B) to the overall mutation load in breast, head/neck, lung, bladder, cervical, ovarian, and other cancer types. However, the complete landscape of mutations attributable to this enzyme has yet to be determined in a controlled human cell system. We report a conditional and isogenic system for A3B induction, genomic DNA deamination, and mutagenesis. Human 293-derived cells were engineered to express doxycycline-inducible A3B-eGFP or eGFP constructs. Cells were subjected to 10 rounds of A3B-eGFP exposure that each caused 80-90% cell death. Control pools were subjected to parallel rounds of non-toxic eGFP exposure, and dilutions were done each round to mimic A3B-eGFP induced population fluctuations. Targeted sequencing of portions of TP53 and MYC demonstrated greater mutation accumulation in the A3B-eGFP exposed pools. Clones were generated and microarray analyses were used to identify those with the greatest number of SNP alterations for whole genome sequencing. A3B-eGFP exposed clones showed global increases in C-to-T transition mutations, enrichments for cytosine mutations within A3B-preferred trinucleotide motifs, and more copy number aberrations. Surprisingly, both control and A3B-eGFP clones also elicited strong mutator phenotypes characteristic of defective mismatch repair. Despite this additional mutational process, the 293-based system characterized here still yielded a genome-wide view of A3B-catalyzed mutagenesis in human cells and a system for additional studies on the compounded effects of simultaneous mutation mechanisms in cancer cells..
Vollan, H.K.
Rueda, O.M.
Chin, S.-.
Curtis, C.
Turashvili, G.
Shah, S.
Lingjærde, O.C.
Yuan, Y.
Ng, C.K.
Dunning, M.J.
Dicks, E.
Provenzano, E.
Sammut, S.
McKinney, S.
Ellis, I.O.
Pinder, S.
Purushotham, A.
Murphy, L.C.
Kristensen, V.N.
METABRIC Group,
Brenton, J.D.
Pharoah, P.D.
Børresen-Dale, A.-.
Aparicio, S.
Caldas, C.
(2015). A tumor DNA complex aberration index is an independent predictor of survival in breast and ovarian cancer. Mol oncol,
Vol.9
(1),
pp. 115-127.
show abstract
full text
Complex focal chromosomal rearrangements in cancer genomes, also called "firestorms", can be scored from DNA copy number data. The complex arm-wise aberration index (CAAI) is a score that captures DNA copy number alterations that appear as focal complex events in tumors, and has potential prognostic value in breast cancer. This study aimed to validate this DNA-based prognostic index in breast cancer and test for the first time its potential prognostic value in ovarian cancer. Copy number alteration (CNA) data from 1950 breast carcinomas (METABRIC cohort) and 508 high-grade serous ovarian carcinomas (TCGA dataset) were analyzed. Cases were classified as CAAI positive if at least one complex focal event was scored. Complex alterations were frequently localized on chromosome 8p (n = 159), 17q (n = 176) and 11q (n = 251). CAAI events on 11q were most frequent in estrogen receptor positive (ER+) cases and on 17q in estrogen receptor negative (ER-) cases. We found only a modest correlation between CAAI and the overall rate of genomic instability (GII) and number of breakpoints (r = 0.27 and r = 0.42, p < 0.001). Breast cancer specific survival (BCSS), overall survival (OS) and ovarian cancer progression free survival (PFS) were used as clinical end points in Cox proportional hazard model survival analyses. CAAI positive breast cancers (43%) had higher mortality: hazard ratio (HR) of 1.94 (95%CI, 1.62-2.32) for BCSS, and of 1.49 (95%CI, 1.30-1.71) for OS. Representations of the 70-gene and the 21-gene predictors were compared with CAAI in multivariable models and CAAI was independently significant with a Cox adjusted HR of 1.56 (95%CI, 1.23-1.99) for ER+ and 1.55 (95%CI, 1.11-2.18) for ER- disease. None of the expression-based predictors were prognostic in the ER- subset. We found that a model including CAAI and the two expression-based prognostic signatures outperformed a model including the 21-gene and 70-gene signatures but excluding CAAI. Inclusion of CAAI in the clinical prognostication tool PREDICT significantly improved its performance. CAAI positive ovarian cancers (52%) also had worse prognosis: HRs of 1.3 (95%CI, 1.1-1.7) for PFS and 1.3 (95%CI, 1.1-1.6) for OS. This study validates CAAI as an independent predictor of survival in both ER+ and ER- breast cancer and reveals a significant prognostic value for CAAI in high-grade serous ovarian cancer..
Tutt, A.
(2015). Targeting DNA repair pathways. Breast,
Vol.24,
pp. S9-S10.
Kunkler, I.H.
Williams, L.J.
Jack, W.J.
Cameron, D.A.
Dixon, J.M.
PRIME II investigators,
(2015). Breast-conserving surgery with or without irradiation in women aged 65 years or older with early breast cancer (PRIME II): a randomised controlled trial. Lancet oncol,
Vol.16
(3),
pp. 266-273.
show abstract
BACKGROUND: For most older women with early breast cancer, standard treatment after breast-conserving surgery is adjuvant whole-breast radiotherapy and adjuvant endocrine treatment. We aimed to assess the effect omission of whole-breast radiotherapy would have on local control in older women at low risk of local recurrence at 5 years. METHODS: Between April 16, 2003, and Dec 22, 2009, 1326 women aged 65 years or older with early breast cancer judged low-risk (ie, hormone receptor-positive, axillary node-negative, T1-T2 up to 3 cm at the longest dimension, and clear margins; grade 3 tumour histology or lymphovascular invasion, but not both, were permitted), who had had breast-conserving surgery and were receiving adjuvant endocrine treatment, were recruited into a phase 3 randomised controlled trial at 76 centres in four countries. Eligible patients were randomly assigned to either whole-breast radiotherapy (40-50 Gy in 15-25 fractions) or no radiotherapy by computer-generated permuted block randomisation, stratified by centre, with a block size of four. The primary endpoint was ipsilateral breast tumour recurrence. Follow-up continues and will end at the 10-year anniversary of the last randomised patient. Analyses were done by intention to treat. The trial is registered on ISRCTN.com, number ISRCTN95889329. FINDINGS: 658 women who had undergone breast-conserving surgery and who were receiving adjuvant endocrine treatment were randomly assigned to receive whole-breast irradiation and 668 were allocated to no further treatment. After median follow-up of 5 years (IQR 3·84-6·05), ipsilateral breast tumour recurrence was 1·3% (95% CI 0·2-2·3; n=5) in women assigned to whole-breast radiotherapy and 4·1% (2·4-5·7; n=26) in those assigned no radiotherapy (p=0·0002). Compared with women allocated to whole-breast radiotherapy, the univariate hazard ratio for ipsilateral breast tumour recurrence in women assigned to no radiotherapy was 5·19 (95% CI 1·99-13·52; p=0·0007). No differences in regional recurrence, distant metastases, contralateral breast cancers, or new breast cancers were noted between groups. 5-year overall survival was 93·9% (95% CI 91·8-96·0) in both groups (p=0·34). 89 women died; eight of 49 patients allocated to no radiotherapy and four of 40 assigned to radiotherapy died from breast cancer. INTERPRETATION: Postoperative whole-breast radiotherapy after breast-conserving surgery and adjuvant endocrine treatment resulted in a significant but modest reduction in local recurrence for women aged 65 years or older with early breast cancer 5 years after randomisation. However, the 5-year rate of ipsilateral breast tumour recurrence is probably low enough for omission of radiotherapy to be considered for some patients. FUNDING: Chief Scientist Office (Scottish Government), Breast Cancer Institute (Western General Hospital, Edinburgh)..
Soady, K.J.
Kendrick, H.
Gao, Q.
Tutt, A.
Zvelebil, M.
Ordonez, L.D.
Quist, J.
Tan, D.W.
Isacke, C.M.
Grigoriadis, A.
Smalley, M.J.
(2015). Mouse mammary stem cells express prognostic markers for triple-negative breast cancer. Breast cancer res,
Vol.17
(1),
p. 31.
show abstract
full text
INTRODUCTION: Triple-negative breast cancer (TNBC) is a heterogeneous group of tumours in which chemotherapy, the current mainstay of systemic treatment, is often initially beneficial but with a high risk of relapse and metastasis. There is currently no means of predicting which TNBC will relapse. We tested the hypothesis that the biological properties of normal stem cells are re-activated in tumour metastasis and that, therefore, the activation of normal mammary stem cell-associated gene sets in primary TNBC would be highly prognostic for relapse and metastasis. METHODS: Mammary basal stem and myoepithelial cells were isolated by flow cytometry and tested in low-dose transplant assays. Gene expression microarrays were used to establish expression profiles of the stem and myoepithelial populations; these were compared to each other and to our previously established mammary epithelial gene expression profiles. Stem cell genes were classified by Gene Ontology (GO) analysis and the expression of a subset analysed in the stem cell population at single cell resolution. Activation of stem cell genes was interrogated across different breast cancer cohorts and within specific subtypes and tested for clinical prognostic power. RESULTS: A set of 323 genes was identified that was expressed significantly more highly in the purified basal stem cells compared to all other cells of the mammary epithelium. A total of 109 out of 323 genes had been associated with stem cell features in at least one other study in addition to our own, providing further support for their involvement in the biology of this cell type. GO analysis demonstrated an enrichment of these genes for an association with cell migration, cytoskeletal regulation and tissue morphogenesis, consistent with a role in invasion and metastasis. Single cell resolution analysis showed that individual cells co-expressed both epithelial- and mesenchymal-associated genes/proteins. Most strikingly, we demonstrated that strong activity of this stem cell gene set in TNBCs identified those tumours most likely to rapidly progress to metastasis. CONCLUSIONS: Our findings support the hypothesis that the biological properties of normal stem cells are drivers of metastasis and that these properties can be used to stratify patients with a highly heterogeneous disease such as TNBC..
Watkins, J.
Weekes, D.
Shah, V.
Gazinska, P.
Joshi, S.
Sidhu, B.
Gillett, C.
Pinder, S.
Vanoli, F.
Jasin, M.
Mayrhofer, M.
Isaksson, A.
Cheang, M.C.
Mirza, H.
Frankum, J.
Lord, C.J.
Ashworth, A.
Vinayak, S.
Ford, J.M.
Telli, M.L.
Grigoriadis, A.
Tutt, A.N.
(2015). Genomic Complexity Profiling Reveals That HORMAD1 Overexpression Contributes to Homologous Recombination Deficiency in Triple-Negative Breast Cancers. Cancer discov,
Vol.5
(5),
pp. 488-505.
show abstract
full text
UNLABELLED: Triple-negative breast cancers (TNBC) are characterized by a wide spectrum of genomic alterations, some of which might be caused by defects in DNA repair processes such as homologous recombination (HR). Despite this understanding, associating particular patterns of genomic instability with response to therapy has been challenging. Here, we show that allelic-imbalanced copy-number aberrations (AiCNA) are more prevalent in TNBCs that respond to platinum-based chemotherapy, thus providing a candidate predictive biomarker for this disease. Furthermore, we show that a high level of AiCNA is linked with elevated expression of a meiosis-associated gene, HORMAD1. Elevated HORMAD1 expression suppresses RAD51-dependent HR and drives the use of alternative forms of DNA repair, the generation of AiCNAs, as well as sensitizing cancer cells to HR-targeting therapies. Our data therefore provide a mechanistic association between HORMAD1 expression, a specific pattern of genomic instability, and an association with response to platinum-based chemotherapy in TNBC. SIGNIFICANCE: Previous studies have shown correlation between mutational "scars" and sensitivity to platinums extending beyond associations with BRCA1/2 mutation, but do not elucidate the mechanism. Here, a novel allele-specific copy-number characterization of genome instability identifies and functionally validates the inappropriate expression of the meiotic gene HORMAD1 as a driver of HR deficiency in TNBC, acting to induce allelic imbalance and moderate platinum and PARP inhibitor sensitivity with implications for the use of such "scars" and expression of meiotic genes as predictive biomarkers..
Ju, Y.S.
Tubio, J.M.
Mifsud, W.
Fu, B.
Davies, H.R.
Ramakrishna, M.
Li, Y.
Yates, L.
Gundem, G.
Tarpey, P.S.
Behjati, S.
Papaemmanuil, E.
Martin, S.
Fullam, A.
Gerstung, M.
ICGC Prostate Cancer Working Group,
ICGC Bone Cancer Working Group,
ICGC Breast Cancer Working Group,
Nangalia, J.
Green, A.R.
Caldas, C.
Borg, Å.
Tutt, A.
Lee, M.T.
van't Veer, L.J.
Tan, B.K.
Aparicio, S.
Span, P.N.
Martens, J.W.
Knappskog, S.
Vincent-Salomon, A.
Børresen-Dale, A.-.
Eyfjörd, J.E.
Myklebost, O.
Flanagan, A.M.
Foster, C.
Neal, D.E.
Cooper, C.
Eeles, R.
Bova, S.G.
Lakhani, S.R.
Desmedt, C.
Thomas, G.
Richardson, A.L.
Purdie, C.A.
Thompson, A.M.
McDermott, U.
Yang, F.
Nik-Zainal, S.
Campbell, P.J.
Stratton, M.R.
(2015). Frequent somatic transfer of mitochondrial DNA into the nuclear genome of human cancer cells. Genome res,
Vol.25
(6),
pp. 814-824.
show abstract
Mitochondrial genomes are separated from the nuclear genome for most of the cell cycle by the nuclear double membrane, intervening cytoplasm, and the mitochondrial double membrane. Despite these physical barriers, we show that somatically acquired mitochondrial-nuclear genome fusion sequences are present in cancer cells. Most occur in conjunction with intranuclear genomic rearrangements, and the features of the fusion fragments indicate that nonhomologous end joining and/or replication-dependent DNA double-strand break repair are the dominant mechanisms involved. Remarkably, mitochondrial-nuclear genome fusions occur at a similar rate per base pair of DNA as interchromosomal nuclear rearrangements, indicating the presence of a high frequency of contact between mitochondrial and nuclear DNA in some somatic cells. Transmission of mitochondrial DNA to the nuclear genome occurs in neoplastically transformed cells, but we do not exclude the possibility that some mitochondrial-nuclear DNA fusions observed in cancer occurred years earlier in normal somatic cells..
Schouten, P.C.
Grigoriadis, A.
Kuilman, T.
Mirza, H.
Watkins, J.A.
Cooke, S.A.
van Dyk, E.
Severson, T.M.
Rueda, O.M.
Hoogstraat, M.
Verhagen, C.V.
Natrajan, R.
Chin, S.-.
Lips, E.H.
Kruizinga, J.
Velds, A.
Nieuwland, M.
Kerkhoven, R.M.
Krijgsman, O.
Vens, C.
Peeper, D.
Nederlof, P.M.
Caldas, C.
Tutt, A.N.
Wessels, L.F.
Linn, S.C.
(2015). Robust BRCA1-like classification of copy number profiles of samples repeated across different datasets and platforms. Mol oncol,
Vol.9
(7),
pp. 1274-1286.
show abstract
Breast cancers with BRCA1 germline mutation have a characteristic DNA copy number (CN) pattern. We developed a test that assigns CN profiles to be 'BRCA1-like' or 'non-BRCA1-like', which refers to resembling a BRCA1-mutated tumor or resembling a tumor without a BRCA1 mutation, respectively. Approximately one third of the BRCA1-like breast cancers have a BRCA1 mutation, one third has hypermethylation of the BRCA1 promoter and one third has an unknown reason for being BRCA1-like. This classification is indicative of patients' response to high dose alkylating and platinum containing chemotherapy regimens, which targets the inability of BRCA1 deficient cells to repair DNA double strand breaks. We investigated whether this classification can be reliably obtained with next generation sequencing and copy number platforms other than the bacterial artificial chromosome (BAC) array Comparative Genomic Hybridization (aCGH) on which it was originally developed. We investigated samples from 230 breast cancer patients for which a CN profile had been generated on two to five platforms, comprising low coverage CN sequencing, CN extraction from targeted sequencing panels (CopywriteR), Affymetrix SNP6.0, 135K/720K oligonucleotide aCGH, Affymetrix Oncoscan FFPE (MIP) technology, 3K BAC and 32K BAC aCGH. Pairwise comparison of genomic position-mapped profiles from the original aCGH platform and other platforms revealed concordance. For most cases, biological differences between samples exceeded the differences between platforms within one sample. We observed the same classification across different platforms in over 80% of the patients and kappa values of at least 0.36. Differential classification could be attributed to CN profiles that were not strongly associated to one class. In conclusion, we have shown that the genomic regions that define our BRCA1-like classifier are robustly measured by different CN profiling technologies, providing the possibility to retro- and prospectively investigate BRCA1-like classification across a wide range of CN platforms..
Di Leo, A.
Curigliano, G.
Diéras, V.
Malorni, L.
Sotiriou, C.
Swanton, C.
Thompson, A.
Tutt, A.
Piccart, M.
(2015). New approaches for improving outcomes in breast cancer in Europe. Breast,
Vol.24
(4),
pp. 321-330.
show abstract
Considerable progress has been made in breast cancer treatment in Europe over the past three decades, yet survival rates for metastatic disease remain poor, underlining the need for further advances. While the use of predictive biomarkers for response to systemic therapy could improve drug development efficiency, progress in identifying such markers has been slow. The currently inadequate classification of breast cancer subtypes is a further challenge. Improved understanding of the molecular pathology of the disease has led to the identification of new targets for drug treatment, and evolving classifications should reflect these developments. Further ongoing challenges include difficulties in finding optimal combinations and sequences of systemic therapies, circumventing multidrug resistance and intra-tumor heterogeneity, problems associated with fragmentation in clinical trials and translational research efforts. Adoption of some of the strategies identified in this article may lead to further improvements in outcomes for patients with the disease..
Coates, A.S.
Winer, E.P.
Goldhirsch, A.
Gelber, R.D.
Gnant, M.
Piccart-Gebhart, M.
Thürlimann, B.
Senn, H.-.
Panel Members,
(2015). Tailoring therapies--improving the management of early breast cancer: St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2015. Ann oncol,
Vol.26
(8),
pp. 1533-1546.
show abstract
The 14th St Gallen International Breast Cancer Conference (2015) reviewed substantial new evidence on locoregional and systemic therapies for early breast cancer. Further experience has supported the adequacy of tumor margins defined as 'no ink on invasive tumor or DCIS' and the safety of omitting axillary dissection in specific cohorts. Radiotherapy trials support irradiation of regional nodes in node-positive disease. Considering subdivisions within luminal disease, the Panel was more concerned with indications for the use of specific therapies, rather than surrogate identification of intrinsic subtypes as measured by multiparameter molecular tests. For the treatment of HER2-positive disease in patients with node-negative cancers up to 1 cm, the Panel endorsed a simplified regimen comprising paclitaxel and trastuzumab without anthracycline as adjuvant therapy. For premenopausal patients with endocrine responsive disease, the Panel endorsed the role of ovarian function suppression with either tamoxifen or exemestane for patients at higher risk. The Panel noted the value of an LHRH agonist given during chemotherapy for premenopausal women with ER-negative disease in protecting against premature ovarian failure and preserving fertility. The Panel noted increasing evidence for the prognostic value of commonly used multiparameter molecular markers, some of which also carried prognostic information for late relapse. The Panel noted that the results of such tests, where available, were frequently used to assist decisions about the inclusion of cytotoxic chemotherapy in the treatment of patients with luminal disease, but noted that threshold values had not been established for this purpose for any of these tests. Multiparameter molecular assays are expensive and therefore unavailable in much of the world. The majority of new breast cancer cases and breast cancer deaths now occur in less developed regions of the world. In these areas, less expensive pathology tests may provide valuable information. The Panel recommendations on treatment are not intended to apply to all patients, but rather to establish norms appropriate for the majority. Again, economic considerations may require that less expensive and only marginally less effective therapies may be necessary in less resourced areas. Panel recommendations do not imply unanimous agreement among Panel members. Indeed, very few of the 200 questions received 100% agreement from the Panel. In the text below, wording is intended to convey the strength of Panel support for each recommendation, while details of Panel voting on each question are available in supplementary Appendix S2, available at Annals of Oncology online..
Tutt, A.
(2015). RESURRECTING PARP INHIBITION. Breast,
Vol.24,
pp. S27-S27.
Johnson, P.
Challis, R.
Chowdhury, F.
Gao, Y.
Harvey, M.
Geldart, T.
Kerr, P.
Chan, C.
Smith, A.
Steven, N.
Edwards, C.
Ashton-Key, M.
Hodges, E.
Tutt, A.
Ottensmeier, C.
Glennie, M.
Williams, A.
(2015). Clinical and biological effects of an agonist anti-CD40 antibody: a Cancer Research UK phase I study. Clin cancer res,
Vol.21
(6),
pp. 1321-1328.
show abstract
PURPOSE: This phase I study aimed to establish the biologic effects and MTD of the agonistic IgG1 chimeric anti-CD40 antibody ChiLob7/4 in patients (pts) with a range of CD40-expressing solid tumors and diffuse large B-cell lymphoma, resistant to conventional therapy. Potential mechanisms of action for agonistic anti-CD40 include direct cytotoxic effects on tumor cells and conditioning of antigen-presenting cells. EXPERIMENTAL DESIGN: ChiLob7/4 was given by IV infusion weekly for 4 doses at a range from 0.5 to 240 mg/dose. Validated ELISAs were used to quantify ChiLob7/4 in serum and test for anti-chimeric MAb (HACA) responses. Pharmacodynamic assessments included quantitation of T-cell, natural killer-cell, and B-cell numbers and activation in blood by flow cytometry and a panel of cytokines in plasma by Luminex technology. Planned dose escalation was in cohorts of 3 patients until MTD or biologic effect, defined as reduction of peripheral blood CD19(+) B cells to 10% or less of baseline. RESULTS: Twenty-nine courses of treatment were given to 28 subjects. The MTD was 200 mg × 4, with dose-limiting toxicity of liver transaminase elevations at 240 mg. At 200 mg (range between 2.1 mg/kg and 3.3 mg/kg based on patient body weight), the trough level pretreatment was above 25 μg/mL. Grade 1-2 infusion reactions were seen above the dose of 16 mg, but could be prevented with single-dose corticosteroid premedication. HACA responses were seen after doses between 1.6 mg and 50 mg, but not above this. There were dose-dependent falls in blood B-cell numbers accompanied by reduced expression of CD21, and transient reductions in NK cell numbers with increased CD54 expression from 50 mg upward. MIP-1β and IL12 plasma concentrations rose after doses above 16 mg. Fifteen of 29 treatments were accompanied by disease stabilization for a median 6 months, the longest for 37 months. CONCLUSIONS: ChiLob7/4 can activate B and NK cells at doses that can be administered safely, and should be tested in combination with other antibodies and chemotherapy agents..
Lord, C.J.
Tutt, A.N.
Ashworth, A.
(2015). Synthetic lethality and cancer therapy: lessons learned from the development of PARP inhibitors. Annu rev med,
Vol.66,
pp. 455-470.
show abstract
The genetic concept of synthetic lethality, in which the combination or synthesis of mutations in multiple genes results in cell death, provides a framework to design novel therapeutic approaches to cancer. Already there are promising indications, from clinical trials exploiting this concept by using poly(ADP-ribose) polymerase (PARP) inhibitors in patients with germline BRCA1 or BRCA2 gene mutations, that this approach could be beneficial. We discuss the biological rationale for BRCA-PARP synthetic lethality, how the synthetic lethal approach is being assessed in the clinic, and how mechanisms of resistance are starting to be dissected. Applying the synthetic lethal concept to target non-BRCA-mutant cancers also has clear potential, and we discuss how some of the principles learned in developing PARP inhibitors might also drive the development of additional genetic approaches..
Tsang, Y.
Ciurlionis, L.
Kirby, A.M.
Locke, I.
Venables, K.
Yarnold, J.R.
Titley, J.
Bliss, J.
Coles, C.E.
IMPORT Trial Management Group,
(2015). Clinical impact of IMPORT HIGH trial (CRUK/06/003) on breast radiotherapy practices in the United Kingdom. Br j radiol,
Vol.88
(1056),
p. 20150453.
show abstract
full text
OBJECTIVE: IMPORT HIGH is a multicentre randomized UK trial testing dose-escalated intensity-modulated radiotherapy (IMRT) after tumour excision in females with early breast cancer and higher than average local recurrence risk. A survey was carried out to investigate the impact of this trial on the adoption of advanced breast radiotherapy (RT) techniques in the UK. METHODS: A questionnaire was sent to all 26 IMPORT HIGH recruiting RT centres to determine whether the trial has influenced non-trial breast RT techniques in terms of volume delineation, dosimetry, treatment delivery and verification. In order to compare the clinical practice of breast RT between IMPORT HIGH and non-IMPORT HIGH centres, parts of the Royal College of Radiologists (RCR) breast RT audit result were used in this study. RESULTS: 26/26 participating centres completed the questionnaire. After joining the trial, the number of centres routinely using tumour bed clips to guide whole-breast RT rose from 5 (19%) to 21 (81%). 20/26 (77%) centres now contour target volumes and organs at risk (OARs) in some or all patients compared with 14 (54%) before the trial. 14/26 (54%) centres offer inverse-planned IMRT for selected non-trial patients with breast cancer, and 10/14 (71%) have adopted the IMPORT HIGH trial protocol for target volume and OARs dose constraints. Only 2/26 (8%) centres used clip information routinely for breast treatment verification prior to IMPORT HIGH, a minority that has since risen to 7/26 (27%). Data on 1386 patients was included from the RCR audit. This suggested that more cases from IMPORT HIGH centres had surgical clips implanted (83 vs 67%), were treated using CT guided planning with full three-dimensional dose compensation (100 vs 75%), and were treated with photon boost RT (30 vs 8%). CONCLUSION: The study suggests that participation in the IMPORT HIGH trial has played an important part in providing the guidance and support networks needed for the safe integration of advanced RT techniques, where appropriate, as a standard of care for breast cancer patients treated at participating cancer centres. ADVANCES IN KNOWLEDGE: We investigated the impact of the IMPORT HIGH trial on the adoption of advanced breast RT techniques in the UK and the trial has influenced non-trial breast RT techniques in terms of volume delineation, dosimetry, treatment delivery and verification..
Marra, P.
Mathew, S.
Grigoriadis, A.
Wu, Y.
Kyle-Cezar, F.
Watkins, J.
Rashid, M.
De Rinaldis, E.
Hessey, S.
Gazinska, P.
Hayday, A.
Tutt, A.
(2014). IL15RA Drives Antagonistic Mechanisms of Cancer Development and Immune Control in Lymphocyte-Enriched Triple-Negative Breast Cancers. Cancer research,
Vol.74
(17),
pp. 4908-4921.
Watkins, J.A.
Irshad, S.
Grigoriadis, A.
Tutt, A.N.
(2014). Genomic scars as biomarkers of homologous recombination deficiency and drug response in breast and ovarian cancers. Breast cancer res,
Vol.16
(3),
p. 211.
show abstract
Poly (ADP-ribose) polymerase (PARP) inhibitors and platinum-based chemotherapies have been found to be particularly effective in tumors that harbor deleterious germline or somatic mutations in the BRCA1 or BRCA2 genes, the products of which contribute to the conservative homologous recombination repair of DNA double-strand breaks. Nonetheless, several setbacks in clinical trial settings have highlighted some of the issues surrounding the investigation of PARP inhibitors, especially the identification of patients who stand to benefit from such drugs. One potential approach to finding this patient subpopulation is to examine the tumor DNA for evidence of a homologous recombination defect. However, although the genomes of many breast and ovarian cancers are replete with aberrations, the presence of numerous factors able to shape the genomic landscape means that only some of the observed DNA abnormalities are the outcome of a cancer cell's inability to faithfully repair DNA double-strand breaks. Consequently, recently developed methods for comprehensively capturing the diverse ways in which homologous recombination deficiencies may arise beyond BRCA1/2 mutation have used DNA microarray and sequencing data to account for potentially confounding features in the genome. Scores capturing telomeric allelic imbalance, loss of heterozygosity (LOH) and large scale transition score, as well as the total number of coding mutations are measures that summarize the total burden of certain forms of genomic abnormality. By contrast, other studies have comprehensively catalogued different types of mutational pattern and their relative contributions to a given tumor sample. Although at least one study to explore the use of the LOH scar in a prospective clinical trial of a PARP inhibitor in ovarian cancer is under way, limitations that result in a relatively low positive predictive value for these biomarkers remain. Tumors whose genome has undergone one or more events that restore high-fidelity homologous recombination are likely to be misclassified as double-strand break repair-deficient and thereby sensitive to PARP inhibitors and DNA damaging chemotherapies as a result of prior repair deficiency and its genomic scarring. Therefore, we propose that integration of a genomic scar-based biomarker with a marker of resistance in a high genomic scarring burden context may improve the performance of any companion diagnostic for PARP inhibitors..
Irshad, S.
Gillett, C.
Pinder, S.E.
A'hern, R.P.
Dowsett, M.
Ellis, I.O.
Bartlett, J.M.
Bliss, J.M.
Hanby, A.
Johnston, S.
Barrett-Lee, P.
Ellis, P.
Tutt, A.
(2014). Assessment of microtubule-associated protein (MAP)-Tau expression as a predictive and prognostic marker in TACT; a trial assessing substitution of sequential docetaxel for FEC as adjuvant chemotherapy for early breast cancer. Breast cancer res treat,
Vol.144
(2),
pp. 331-341.
show abstract
The TACT trial is the largest study assessing the benefit of taxanes as part of adjuvant therapy for early breast cancer. The goal of this translational study was to clarify the predictive and prognostic value of Tau within the TACT trial. Tissue microarrays (TMA) were available from 3,610 patients. ER, PR, HER2 from the TACT trial and Tau protein expression was determined by immunohistochemistry on duplicate TMAs. Two parallel scoring systems were generated for Tau expression ('dichotomised' vs. 'combined' score). The positivity rate of Tau expression was 50 % in the trial population (n = 2,483). Tau expression correlated positively with ER (p < 0.001) and PR status (p < 0.001); but negatively with histological grade (p < 0.001) and HER2 status (p < 0.001). Analyses with either scoring systems for Tau expression demonstrated no significant interaction between Tau expression and efficacy of docetaxel. Contrary to the hypothesis that taxane benefit would be enriched in Tau negative/low patients, the only groups with a suggestion of a reduced event rate in the taxane group were the HER2-positive, Tau positive subgroups. Tau expression was seen to be a prognostic factor on univariate analysis associated with an improved DFS, independent of the treatment group (p < 0.001). It had no prognostic value in ER-negative tumours and the weak prognostic effect of Tau in ER-positive tumours (p = 0.02) diminished, when considering ER as an ordinal variable. On multivariable analyses, Tau had no prognostic value in either group. In addition, no significant interaction between Tau expression and benefit from docetaxel in patients within the PR-positive and negative subsets was seen. This is now the second large adjuvant study, and the first with quantitative analysis of ER and Tau expression, failing to show an association between Tau and taxane benefit with limited utility as a prognostic marker for Tau in ER-positive early breast cancer patients..
Parikh, J.
Selmi, M.
Charles-Edwards, G.
Glendenning, J.
Ganeshan, B.
Verma, H.
Mansi, J.
Harries, M.
Tutt, A.
Goh, V.
(2014). Changes in Primary Breast Cancer Heterogeneity May Augment Midtreatment MR Imaging Assessment of Response to Neoadjuvant Chemotherapy. Radiology,
Vol.272
(1),
pp. 100-112.
Vantourout, P.
Willcox, C.
Turner, A.
Swanson, C.M.
Haque, Y.
Sobolev, O.
Grigoriadis, A.
Tutt, A.
Hayday, A.
(2014). Immunological Visibility: Posttranscriptional Regulation of Human NKG2D Ligands by the EGF Receptor Pathway. Science translational medicine,
Vol.6
(231).
full text
Kyle, F.
Hessey, S.
Wu, Y.
Tutt, A.
Hayday, A.
(2014). Characterising the immune landscape of healthy breast tissue and breast tumours in humans. Immunology,
Vol.143,
pp. 144-144.
Kiuchi, T.
Ortiz-Zapater, E.
Monypenny, J.
Matthews, D.R.
Nguyen, L.K.
Barbeau, J.
Coban, O.
Lawler, K.
Burford, B.
Rolfe, D.J.
de Rinaldis, E.
Dafou, D.
Simpson, M.A.
Woodman, N.
Pinder, S.
Gillett, C.E.
Devauges, V.
Poland, S.P.
Fruhwirth, G.
Marra, P.
Boersma, Y.L.
Plueckthun, A.
Gullick, W.J.
Yarden, Y.
Santis, G.
Winn, M.
Kholodenko, B.N.
Martin-Fernandez, M.L.
Parker, P.
Tutt, A.
Ameer-Beg, S.M.
Ng, T.
(2014). The ErbB4 CYT2 variant protects EGFR from ligand-induced degradation to enhance cancer cell motility. Science signaling,
Vol.7
(339).
Eccles, S.A.
Aboagye, E.O.
Ali, S.
Anderson, A.S.
Armes, J.
Berditchevski, F.
Blaydes, J.P.
Brennan, K.
Brown, N.J.
Bryant, H.E.
Bundred, N.J.
Burchell, J.M.
Campbell, A.M.
Carroll, J.S.
Clarke, R.B.
Coles, C.E.
Cook, G.J.
Cox, A.
Curtin, N.J.
Dekker, L.V.
Silva, I.D.
Duffy, S.W.
Easton, D.F.
Eccles, D.M.
Edwards, D.R.
Edwards, J.
Evans, D.
Fenlon, D.F.
Flanagan, J.M.
Foster, C.
Gallagher, W.M.
Garcia-Closas, M.
Gee, J.M.
Gescher, A.J.
Goh, V.
Groves, A.M.
Harvey, A.J.
Harvie, M.
Hennessy, B.T.
Hiscox, S.
Holen, I.
Howell, S.J.
Howell, A.
Hubbard, G.
Hulbert-Williams, N.
Hunter, M.S.
Jasani, B.
Jones, L.J.
Key, T.J.
Kirwan, C.C.
Kong, A.
Kunkler, I.H.
Langdon, S.P.
Leach, M.O.
Mann, D.J.
Marshall, J.F.
Martin, L.
Martin, S.G.
Macdougall, J.E.
Miles, D.W.
Miller, W.R.
Morris, J.R.
Moss, S.M.
Mullan, P.
Natrajan, R.
O'Connor, J.P.
O'Connor, R.
Palmieri, C.
Pharoah, P.D.
Rakha, E.A.
Reed, E.
Robinson, S.P.
Sahai, E.
Saxton, J.M.
Schmid, P.
Smalley, M.J.
Speirs, V.
Stein, R.
Stingl, J.
Streuli, C.H.
Tutt, A.N.
Velikova, G.
Walker, R.A.
Watson, C.J.
Williams, K.J.
Young, L.S.
Thompson, A.M.
(2013). Critical research gaps and translational priorities for the successful prevention and treatment of breast cancer. Breast cancer res,
Vol.15
(5),
p. R92.
show abstract
INTRODUCTION: Breast cancer remains a significant scientific, clinical and societal challenge. This gap analysis has reviewed and critically assessed enduring issues and new challenges emerging from recent research, and proposes strategies for translating solutions into practice. METHODS: More than 100 internationally recognised specialist breast cancer scientists, clinicians and healthcare professionals collaborated to address nine thematic areas: genetics, epigenetics and epidemiology; molecular pathology and cell biology; hormonal influences and endocrine therapy; imaging, detection and screening; current/novel therapies and biomarkers; drug resistance; metastasis, angiogenesis, circulating tumour cells, cancer 'stem' cells; risk and prevention; living with and managing breast cancer and its treatment. The groups developed summary papers through an iterative process which, following further appraisal from experts and patients, were melded into this summary account. RESULTS: The 10 major gaps identified were: (1) understanding the functions and contextual interactions of genetic and epigenetic changes in normal breast development and during malignant transformation; (2) how to implement sustainable lifestyle changes (diet, exercise and weight) and chemopreventive strategies; (3) the need for tailored screening approaches including clinically actionable tests; (4) enhancing knowledge of molecular drivers behind breast cancer subtypes, progression and metastasis; (5) understanding the molecular mechanisms of tumour heterogeneity, dormancy, de novo or acquired resistance and how to target key nodes in these dynamic processes; (6) developing validated markers for chemosensitivity and radiosensitivity; (7) understanding the optimal duration, sequencing and rational combinations of treatment for improved personalised therapy; (8) validating multimodality imaging biomarkers for minimally invasive diagnosis and monitoring of responses in primary and metastatic disease; (9) developing interventions and support to improve the survivorship experience; (10) a continuing need for clinical material for translational research derived from normal breast, blood, primary, relapsed, metastatic and drug-resistant cancers with expert bioinformatics support to maximise its utility. The proposed infrastructural enablers include enhanced resources to support clinically relevant in vitro and in vivo tumour models; improved access to appropriate, fully annotated clinical samples; extended biomarker discovery, validation and standardisation; and facilitated cross-discipline working. CONCLUSIONS: With resources to conduct further high-quality targeted research focusing on the gaps identified, increased knowledge translating into improved clinical care should be achievable within five years..
Marlow, R.
Honeth, G.
Lombardi, S.
Cariati, M.
Hessey, S.
Pipili, A.
Mariotti, V.
Buchupalli, B.
Foster, K.
Bonnet, D.
Grigoriadis, A.
Rameshwar, P.
Purushotham, A.
Tutt, A.
Dontu, G.
(2013). A Novel Model of Dormancy for Bone Metastatic Breast Cancer Cells. Cancer research,
Vol.73
(23),
pp. 6886-6899.
Jack, R.H.
Davies, E.A.
Renshaw, C.
Tutt, A.
Grocock, M.J.
Coupland, V.H.
Moller, H.
(2013). Differences in breast cancer hormone receptor status in ethnic groups: A London population. European journal of cancer,
Vol.49
(3),
pp. 696-702.
Alexandrov, L.B.
Nik-Zainal, S.
Wedge, D.C.
Aparicio, S.A.
Behjati, S.
Biankin, A.V.
Bignell, G.R.
Bolli, N.
Borg, A.
Børresen-Dale, A.-.
Boyault, S.
Burkhardt, B.
Butler, A.P.
Caldas, C.
Davies, H.R.
Desmedt, C.
Eils, R.
Eyfjörd, J.E.
Foekens, J.A.
Greaves, M.
Hosoda, F.
Hutter, B.
Ilicic, T.
Imbeaud, S.
Imielinski, M.
Jäger, N.
Jones, D.T.
Jones, D.
Knappskog, S.
Kool, M.
Lakhani, S.R.
López-Otín, C.
Martin, S.
Munshi, N.C.
Nakamura, H.
Northcott, P.A.
Pajic, M.
Papaemmanuil, E.
Paradiso, A.
Pearson, J.V.
Puente, X.S.
Raine, K.
Ramakrishna, M.
Richardson, A.L.
Richter, J.
Rosenstiel, P.
Schlesner, M.
Schumacher, T.N.
Span, P.N.
Teague, J.W.
Totoki, Y.
Tutt, A.N.
Valdés-Mas, R.
van Buuren, M.M.
van 't Veer, L.
Vincent-Salomon, A.
Waddell, N.
Yates, L.R.
Australian Pancreatic Cancer Genome Initiative,
ICGC Breast Cancer Consortium,
ICGC MMML-Seq Consortium,
ICGC PedBrain,
Zucman-Rossi, J.
Futreal, P.A.
McDermott, U.
Lichter, P.
Meyerson, M.
Grimmond, S.M.
Siebert, R.
Campo, E.
Shibata, T.
Pfister, S.M.
Campbell, P.J.
Stratton, M.R.
(2013). Signatures of mutational processes in human cancer. Nature,
Vol.500
(7463),
pp. 415-421.
show abstract
All cancers are caused by somatic mutations; however, understanding of the biological processes generating these mutations is limited. The catalogue of somatic mutations from a cancer genome bears the signatures of the mutational processes that have been operative. Here we analysed 4,938,362 mutations from 7,042 cancers and extracted more than 20 distinct mutational signatures. Some are present in many cancer types, notably a signature attributed to the APOBEC family of cytidine deaminases, whereas others are confined to a single cancer class. Certain signatures are associated with age of the patient at cancer diagnosis, known mutagenic exposures or defects in DNA maintenance, but many are of cryptic origin. In addition to these genome-wide mutational signatures, hypermutation localized to small genomic regions, 'kataegis', is found in many cancer types. The results reveal the diversity of mutational processes underlying the development of cancer, with potential implications for understanding of cancer aetiology, prevention and therapy..
Yau, C.
Sninsky, J.
Kwok, S.
Wang, A.
Degnim, A.
Ingle, J.N.
Gillett, C.
Tutt, A.
Waldman, F.
Moore, D.
Esserman, L.
Benz, C.C.
(2013). An optimized five-gene multi-platform predictor of hormone receptor negative and triple negative breast cancer metastatic risk. Breast cancer research,
Vol.15
(5).
Kaufmann, M.
von Minckwitz, G.
Mamounas, E.P.
Cameron, D.
Carey, L.A.
Cristofanilli, M.
Denkert, C.
Eiermann, W.
Gnant, M.
Harris, J.R.
Karn, T.
Liedtke, C.
Mauri, D.
Rouzier, R.
Ruckhaeberle, E.
Semiglazov, V.
Symmans, W.F.
Tutt, A.
Pusztai, L.
(2012). Recommendations from an International Consensus Conference on the Current Status and Future of Neoadjuvant Systemic Therapy in Primary Breast Cancer. Annals of surgical oncology,
Vol.19
(5),
pp. 1508-1516.
Vichapat, V.
Garmo, H.
Holmberg, L.
Fentiman, I.S.
Tutt, A.
Gillett, C.
Luechtenborg, M.
(2012). Patterns of metastasis in women with metachronous contralateral breast cancer. British journal of cancer,
Vol.107
(2),
pp. 221-223.
Tutt, A.
(2012). The biology of BRCA-associated breast cancer. Breast,
Vol.21,
pp. S2-S2.
Robertson, L.
Hanson, H.
Seal, S.
Warren-Perry, M.
Hughes, D.
Howell, I.
Turnbull, C.
Houlston, R.
Shanley, S.
Butler, S.
Evans, D.G.
Ross, G.
Eccles, D.
Tutt, A.
Rahman, N.
TNT Trial TMG,
BCSC (UK),
(2012). BRCA1 testing should be offered to individuals with triple-negative breast cancer diagnosed below 50 years. Br j cancer,
Vol.106
(6),
pp. 1234-1238.
show abstract
BACKGROUND: Triple-negative (TN) tumours are the predominant breast cancer subtype in BRCA1 mutation carriers. Recently, it was proposed that all individuals below 50 years of age with TN breast cancer should be offered BRCA testing. We have evaluated the BRCA1 mutation frequency and the implications for clinical practice of undertaking genetic testing in women with TN breast cancer. METHODS: We undertook BRCA1 mutation analysis in 308 individuals with TN breast cancer, 159 individuals from unselected series of breast cancer and 149 individuals from series ascertained on the basis of young age and/or family history. RESULTS: BRCA1 mutations were present in 45 out of 308 individuals. Individuals with TN cancer <50 years had >10% likelihood of carrying a BRCA1 mutation in both the unselected (11 out of 58, 19%) and selected (26 out of 111, 23%) series. However, over a third would not have been offered testing using existing criteria. We estimate that testing all individuals with TN breast cancer <50 years would generate an extra 1200 tests annually in England. CONCLUSION: Women with TN breast cancer diagnosed below 50 years have >10% likelihood of carrying a BRCA1 mutation and are therefore eligible for testing in most centres. However, implementation may place short-term logistical and financial burdens on genetic services..
Turner, N.C.
Tutt, A.N.
(2012). Platinum chemotherapy for BRCA1-related breast cancer: do we need more evidence?. Breast cancer res,
Vol.14
(6),
p. 115.
show abstract
A recent prospective clinical trial provides further evidence that breast cancers arising in germline BRCA1 mutation carriers are highly sensitive to cisplatin chemotherapy. The potential significance of these data for the management of patients with BRCA1-related and BRCA2-related breast cancer is discussed..
Grigoriadis, A.
Mackay, A.
Noel, E.
Wu, P.J.
Natrajan, R.
Frankum, J.
Reis-Filho, J.S.
Tutt, A.
(2012). Molecular characterisation of cell line models for triple-negative breast cancers. Bmc genomics,
Vol.13.
Marlow, R.
Cariati, M.
Honeth, G.M.
Buchupalli, B.
Lombardi, S.
Foster, K.
Bonnet, D.
Purushotham, A.
Tutt, A.
Dontu, G.
(2012). Modeling the breast cancer metastatic niche. Cancer research,
Vol.72.
Metzger-Filho, O.
Tutt, A.
de Azambuja, E.
Saini, K.S.
Viale, G.
Loi, S.
Bradbury, I.
Bliss, J.M.
Azim, H.A.
Ellis, P.
Di Leo, A.
Baselga, J.
Sotiriou, C.
Piccart-Gebhart, M.
(2012). Dissecting the heterogeneity of triple-negative breast cancer. J clin oncol,
Vol.30
(15),
pp. 1879-1887.
show abstract
Triple-negative breast cancer (TNBC) accounts for 15% to 20% of breast cancers. It is a heterogeneous disease, not only on the molecular level, but also on the pathologic and clinical levels. TNBC is associated with a significantly higher probability of relapse and poorer overall survival in the first few years after diagnosis when compared with other breast cancer subtypes. This is observed despite its usual high sensitivity to chemotherapy. In the advanced setting, responses observed with chemotherapy lack durability. Early-stage clinical studies suggested impressive potential when a poly (ADP-ribose) polymerase (PARP) inhibitor is given for the treatment of advanced TNBC with BRCA gene dysfunction. The molecular complexity of TNBC has led to proposed subclassifications, which will be of great value for the development of targeted therapies. In this review, we discuss the biology of TNBC at the pathologic and the molecular levels. We also elaborate on the role of systemic therapies and the results of the first phase III clinical trial evaluating the addition of iniparib, a novel investigational anticancer agent that does not possess characteristics typical of the PARP inhibitor class, in combination with chemotherapy in advanced TNBC..
Stephens, P.J.
Tarpey, P.S.
Davies, H.
Van Loo, P.
Greenman, C.
Wedge, D.C.
Nik-Zainal, S.
Martin, S.
Varela, I.
Bignell, G.R.
Yates, L.R.
Papaemmanuil, E.
Beare, D.
Butler, A.
Cheverton, A.
Gamble, J.
Hinton, J.
Jia, M.
Jayakumar, A.
Jones, D.
Latimer, C.
Lau, K.W.
McLaren, S.
McBride, D.J.
Menzies, A.
Mudie, L.
Raine, K.
Rad, R.
Chapman, M.S.
Teague, J.
Easton, D.
Langerod, A.
Lee, M.T.
Shen, C.-.
Tee, B.T.
Huimin, B.W.
Broeks, A.
Vargas, A.C.
Turashvili, G.
Martens, J.
Fatima, A.
Miron, P.
Chin, S.-.
Thomas, G.
Boyault, S.
Mariani, O.
Lakhani, S.R.
van de Vijver, M.
Van 'tVeer, L.
Foekens, J.
Desmedt, C.
Sotiriou, C.
Tutt, A.
Caldas, C.
Reis-Filho, J.S.
Aparicio, S.A.
Salomon, A.V.
Borresen-Dale, A.-.
Richardson, A.L.
Campbell, P.J.
Futreal, P.A.
Stratton, M.R.
OSBREAC,
(2012). The landscape of cancer genes and mutational processes in breast cancer. Nature,
Vol.486
(7403),
pp. 400-+.
Nik-Zainal, S.
Van Loo, P.
Wedge, D.C.
Alexandrov, L.B.
Greenman, C.D.
Lau, K.W.
Raine, K.
Jones, D.
Marshall, J.
Ramakrishna, M.
Shlien, A.
Cooke, S.L.
Hinton, J.
Menzies, A.
Stebbings, L.A.
Leroy, C.
Jia, M.
Rance, R.
Mudie, L.J.
Gamble, S.J.
Stephens, P.J.
McLaren, S.
Tarpey, P.S.
Papaemmanuil, E.
Davies, H.R.
Varela, I.
McBride, D.J.
Bignell, G.R.
Leung, K.
Butler, A.P.
Teague, J.W.
Martin, S.
Joensson, G.
Mariani, O.
Boyault, S.
Miron, P.
Fatima, A.
Langerod, A.
Aparicio, S.A.
Tutt, A.
Sieuwerts, A.M.
Borg, A.
Thomas, G.
Salomon, A.V.
Richardson, A.L.
Borresen-Dale, A.-.
Futreal, P.A.
Stratton, M.R.
Campbell, P.J.
Consortium, I.C.
(2012). The Life History of 21 Breast Cancers. Cell,
Vol.149
(5).
Nik-Zainal, S.
Alexandrov, L.B.
Wedge, D.C.
Van Loo, P.
Greenman, C.D.
Raine, K.
Jones, D.
Hinton, J.
Marshall, J.
Stebbings, L.A.
Menzies, A.
Martin, S.
Leung, K.
Chen, L.
Leroy, C.
Ramakrishna, M.
Rance, R.
Lau, K.W.
Mudie, L.J.
Varela, I.
McBride, D.J.
Bignell, G.R.
Cooke, S.L.
Shlien, A.
Gamble, J.
Whitmore, I.
Maddison, M.
Tarpey, P.S.
Davies, H.R.
Papaemmanuil, E.
Stephens, P.J.
McLaren, S.
Butler, A.P.
Teague, J.W.
Jonsson, G.
Garber, J.E.
Silver, D.
Miron, P.
Fatima, A.
Boyault, S.
Langerod, A.
Tutt, A.
Martens, J.W.
Aparicio, S.A.
Borg, A.
Salomon, A.V.
Thomas, G.
Borresen-Dale, A.-.
Richardson, A.L.
Neuberger, M.S.
Futreal, P.A.
Campbell, P.J.
Stratton, M.R.
Consortium, I.C.
(2012). Mutational Processes Molding the Genomes of 21 Breast Cancers. Cell,
Vol.149
(5),
pp. 979-993.
Irshad, S.
Grigoriadis, A.
Lawler, K.
Ng, T.
Tutt, A.
(2012). Profiling the Immune Stromal Interface in Breast Cancer and Its Potential for Clinical Impact. Breast care,
Vol.7
(4),
pp. 273-280.
Tutt, A.
(2011). PARP inhibitors - walking them through to adjuvant. Breast,
Vol.20,
pp. S1-S1.
Balmana, J.
Domchek, S.M.
Tutt, A.
Garber, J.E.
(2011). Stumbling Blocks on the Path to Personalized Medicine in Breast Cancer: The Case of PARP Inhibitors for BRCA1/2-Associated Cancers. Cancer discovery,
Vol.1
(1),
pp. 29-34.
Irshad, S.
Ashworth, A.
Tutt, A.
(2011). Therapeutic potential of PARP inhibitors for metastatic breast cancer. Expert review of anticancer therapy,
Vol.11
(8),
pp. 1243-1251.
Vichapat, V.
Gillett, C.
Fentiman, I.S.
Tutt, A.
Holmberg, L.
Luechtenborg, M.
(2011). Risk factors for metachronous contralateral breast cancer suggest two aetiological pathways. European journal of cancer,
Vol.47
(13),
pp. 1919-1927.
Tutt, A.
(2011). ROLE OF PARP INHIBITORS IN TN ADVANCED BREAST CANCER. Breast,
Vol.20,
pp. S17-S17.
Glendenning, J.
Tutt, A.
(2011). PARP inhibitors - current status and the walk towards early breast cancer. Breast,
Vol.20,
pp. S12-S19.
Domchek, S.M.
Mitchell, G.
Lindeman, G.J.
Tung, N.M.
Balmana, J.
Isakoff, S.J.
Schmutzler, R.
Audeh, M.W.
Loman, N.
Scott, C.
Friedlander, M.
Kaufman, B.
Garber, J.E.
Tutt, A.
Robson, M.E.
(2011). Challenges to the Development of New Agents for Molecularly Defined Patient Subsets: Lessons From BRCA1/2-Associated Breast Cancer. Journal of clinical oncology,
Vol.29
(32),
pp. 4224-4226.
Irshad, S.
Ellis, P.
Tutt, A.
(2011). Molecular heterogeneity of triple-negative breast cancer and its clinical implications. Current opinion in oncology,
Vol.23
(6),
pp. 566-577.
Vichapat, V.
Garmo, H.
Holmberg, L.
Fentiman, I.S.
Tutt, A.
Gillett, C.
Luechtenborg, M.
(2011). Prognosis of metachronous contralateral breast cancer: importance of stage, age and interval time between the two diagnoses. Breast cancer research and treatment,
Vol.130
(2),
pp. 609-618.
Julien, S.
Ivetic, A.
Grigoriadis, A.
Qize, D.
Burford, B.
Sproviero, D.
Picco, G.
Gillett, C.
Papp, S.L.
Schaffer, L.
Tutt, A.
Taylor-Papadimitriou, J.
Pinder, S.E.
Burchell, J.M.
(2011). Selectin Ligand Sialyl-Lewis x Antigen Drives Metastasis of Hormone-Dependent Breast Cancers. Cancer research,
Vol.71
(24),
pp. 7683-7693.
full text
Tutt, A.
(2011). Synthetic lethality: Targeting DNA repair mechanisms in clinical trials. Cancer research,
Vol.71.
Jiao, Y.
Lawler, K.
Patel, G.S.
Purushotham, A.
Jones, A.F.
Grigoriadis, A.
Tutt, A.
Ng, T.
Teschendorff, A.E.
(2011). DART: Denoising Algorithm based on Relevance network Topology improves molecular pathway activity inference. Bmc bioinformatics,
Vol.12.
Basu, B.
Ang, J.E.
Crawley, D.
Folkerd, E.
Sarker, D.
Blanco-Codesido, M.
Moran, K.
Wan, S.
Dobbs, N.
Raynaud, F.
Johnston, S.R.
Dowsett, M.
Tutt, A.N.
Spicer, J.F.
Swanton, C.
De Bono, J.S.
(2011). Phase I study of abiraterone acetate (AA) in patients (pts) with estrogen receptor- (ER) or androgen receptor (AR) -positive advanced breast carcinoma resistant to standard endocrine therapies. J clin oncol,
Vol.29
(15_suppl),
p. 2525.
show abstract
2525 Background: Many advanced ER+ breast cancer pts show intrinsic resistance to endocrine treatment with the remainder acquiring resistance. Androgenic steroids upstream of aromatase can drive steroid receptor signaling critical to tumour growth. Evidence also exists for an ERα-/AR+ subset of breast cancers transcriptionally similar to ERα+ disease. We hypothesized that AA, a cytochrome (CYP) 17 inhibitor that irreversibly inhibits androgen and estrogen synthesis, would have anti-tumour activity in ER+ or ER-/AR+ pts. METHODS: Post-menopausal women with ER+ or ER-/AR+ advanced breast cancer resistant to >2 lines of hormone therapies were treated in 6-pt cohorts with AA at doses between 250 and 2000mg daily. RESULTS: A total of 25 pts were treated in the phase I trial with 2 pts ongoing on study. Median time on treatment was 1.8 mths (range 0.7 - 11.6 mths). There were no dose limiting toxicities. The majority of adverse events (AEs) were Common Toxicity Criteria grade 1 or 2: fatigue, nausea, anorexia, dyspnoea, palpitations, dizziness and flushes. Hypokalemia was frequent, due to secondary mineralocorticoid syndrome: grade 3/4 hypokalemia occurred in 4 pts. This was effectively managed with potassium supplementation, hydrocortisone (20 mg mane, 10 mg nocte) and eplerenone (50-200 mg) with no clinical consequences. Other grade 3 AEs were neutropenia and reduced left ventricular ejection fraction (1 pt) and exercise-induced hypotension with dizziness (1 pt). At the 2000mg dose 5/5 subjects had suppression of estradiol, testosterone, DHEA and DHEAS to below the lower limit of detection of the assay. Two pts (both ER+/AR+) continued on study beyond 11 mths, one of whom achieved a radiological partial response and 80% reduction in serum CA15.3 from baseline. Pharmacokinetic data will be presented. CONCLUSIONS: AA is well tolerated in advanced breast cancer pts with preliminary evidence of antitumour activity. The predominant AEs are mechanism-based (hypokalemia) and can be managed expectantly, although careful monitoring of potassium levels is recommended..
Basu, B.
Ang, J.E.
Crawley, D.
Folkerd, E.
Sarker, D.
Blanco-Codesido, M.
Moran, K.
Wan, S.
Dobbs, N.
Raynaud, F.
Johnston, S.R.
Dowsett, M.
Tutt, A.N.
Spicer, J.F.
Swanton, C.
De Bono, J.S.
(2011). Phase I study of abiraterone acetate (AA) in patients (pts) with estrogen receptor-(ER) or androgen receptor (AR)-positive advanced breast carcinoma resistant to standard endocrine therapies. Journal of clinical oncology,
Vol.29
(15).
Turner, N.
Pearson, A.
Sharpe, R.
Lambros, M.
Geyer, F.
Lopez-Garcia, M.A.
Natrajan, R.
Marchio, C.
Iorns, E.
Mackay, A.
Gillett, C.
Grigoriadis, A.
Tutt, A.
Reis-Filho, J.S.
Ashworth, A.
(2010). FGFR1 Amplification Drives Endocrine Therapy Resistance and Is a Therapeutic Target in Breast Cancer. Cancer research,
Vol.70
(5),
pp. 2085-10.
Yap, T.A.
de Bono, J.S.
Kaye, S.B.
Tutt, A.
Ashworth, A.
Schellens, J.H.
(2010). Reply to J Veeck et al. J clin oncol,
.
Molyneux, G.
Geyer, F.C.
Magnay, F.-.
McCarthy, A.
Kendrick, H.
Natrajan, R.
Mackay, A.
Grigoriadis, A.
Tutt, A.
Ashworth, A.
Reis-Filho, J.S.
Smalley, M.J.
(2010). BRCA1 basal-like breast cancers originate from luminal epithelial progenitors and not from basal stem cells. Cell stem cell,
Vol.7
(3),
pp. 403-417.
show abstract
Breast cancers in BRCA1 mutation carriers frequently have a distinctive basal-like phenotype. It has been suggested that this results from an origin in basal breast epithelial stem cells. Here, we demonstrate that deleting Brca1 in mouse mammary epithelial luminal progenitors produces tumors that phenocopy human BRCA1 breast cancers. They also resemble the majority of sporadic basal-like breast tumors. However, directing Brca1 deficiency to basal cells generates tumors that express molecular markers of basal breast cancers but do not histologically resemble either human BRCA1 or the majority of sporadic basal-like breast tumors. These findings support a derivation of the majority of human BRCA1-associated and sporadic basal-like tumors from luminal progenitors rather than from basal stem cells. They also demonstrate that when target cells for transformation have the potential for phenotypic plasticity, tumor phenotypes may not directly reflect histogenesis. This has important implications for cancer prevention strategies..
Stebbing, J.
Ellis, P.
Tutt, A.
(2010). PARP inhibitors in BRCA1-/BRCA2-associated and triple-negative breast cancers. Future oncology,
Vol.6
(4),
pp. 485-486.
Pichert, G.
Jacobs, C.
Jacobs, I.
Menon, U.
Manchanda, R.
Johnson, M.
Hamed, H.
Firth, C.
Evison, M.
Tutt, A.
de Silva, L.
Langman, C.
Izatt, L.
(2010). Novel one-stop multidisciplinary follow-up clinic significantly improves cancer risk management in BRCA1/2 carriers. Familial cancer,
Vol.9
(3),
pp. 313-319.
Cazet, A.
Lefebvre, J.
Adriaenssens, E.
Julien, S.
Bobowski, M.
Grigoriadis, A.
Tutt, A.
Tulasne, D.
Le Bourhis, X.
Delannoy, P.
(2010). G(D3) Synthase Expression Enhances Proliferation and Tumor Growth of MDA-MB-231 Breast Cancer Cells through c-Met Activation. Molecular cancer research,
Vol.8
(11),
pp. 1526-1535.
Fong, P.C.
Yap, T.A.
Boss, D.S.
Carden, C.P.
Mergui-Roelvink, M.
Gourley, C.
De Greve, J.
Lubinski, J.
Shanley, S.
Messiou, C.
A'Hern, R.
Tutt, A.
Ashworth, A.
Stone, J.
Carmichael, J.
Schellens, J.H.
de Bono, J.S.
Kaye, S.B.
(2010). Poly(ADP)-ribose polymerase inhibition: frequent durable responses in BRCA carrier ovarian cancer correlating with platinum-free interval. J clin oncol,
Vol.28
(15),
pp. 2512-2519.
show abstract
PURPOSE: Selective tumor cell cytotoxicity can be achieved through a synthetic lethal strategy using poly(ADP)-ribose polymerase (PARP) inhibitor therapy in BRCA1/2 mutation carriers in whom tumor cells have defective homologous recombination (HR) DNA repair. Platinum-based chemotherapy responses correlate with HR DNA repair capacity. Olaparib is a potent, oral PARP inhibitor that is well tolerated, with antitumor activity in BRCA1/2 mutation carriers. PATIENTS AND METHODS: Patients with BRCA1/2-mutated ovarian cancer were treated with olaparib within a dose-escalation and single-stage expansion of a phase I trial. Antitumor activity was subsequently correlated with platinum sensitivity. RESULTS: Fifty patients were treated: 48 had germline BRCA1/2 mutations; one had a BRCA2 germline sequence change of unknown significance, and another had a strong family history of BRCA1/2-associated cancers who declined mutation testing. Of the 50 patients, 13 had platinum-sensitive disease, 24 had platinum-resistant disease, and 13 had platinum-refractory disease (according to platinum-free interval). Twenty (40%; 95% CI, 26% to 55%) achieved Response Evaluation Criteria in Solid Tumors (RECIST) complete or partial responses and/or tumor marker (CA125) responses, and three (6.0%) maintained RECIST disease stabilization for more than 4 months, giving an overall clinical benefit rate of 46% (95% CI, 32% to 61%). Median response duration was 28 weeks. There was a significant association between the clinical benefit rate and platinum-free interval across the platinum-sensitive, resistant, and refractory subgroups (69%, 45%, and 23%, respectively). Post hoc analyses indicated associations between platinum sensitivity and extent of olaparib response (radiologic change, P = .001; CA125 change, P = .002). CONCLUSION: Olaparib has antitumor activity in BRCA1/2 mutation ovarian cancer, which is associated with platinum sensitivity..
Uva, P.
Lahm, A.
Sbardellati, A.
Grigoriadis, A.
Tutt, A.
de Rinaldis, E.
(2010). Comparative Membranome Expression Analysis in Primary Tumors and Derived Cell Lines. Plos one,
Vol.5
(7).
Tutt, A.
Robson, M.
Garber, J.E.
Domchek, S.M.
Audeh, M.W.
Weitzel, J.N.
Friedlander, M.
Arun, B.
Loman, N.
Schmutzler, R.K.
Wardley, A.
Mitchell, G.
Earl, H.
Wickens, M.
Carmichael, J.
(2010). Oral poly(ADP-ribose) polymerase inhibitor olaparib in patients with BRCA1 or BRCA2 mutations and advanced breast cancer: a proof-of-concept trial. Lancet,
Vol.376
(9737),
pp. 235-244.
Audeh, M.W.
Carmichael, J.
Penson, R.T.
Friedlander, M.
Powell, B.
Bell-McGuinn, K.M.
Scott, C.
Weitzel, J.N.
Oaknin, A.
Loman, N.
Lu, K.
Schmutzler, R.K.
Matulonis, U.
Wickens, M.
Tutt, A.
(2010). Oral poly(ADP-ribose) polymerase inhibitor olaparib in patients with BRCA1 or BRCA2 mutations and recurrent ovarian cancer: a proof-of-concept trial. Lancet,
Vol.376
(9737),
pp. 245-251.
Iverson, A.A.
Gillett, C.
Cane, P.
Santini, C.D.
Vess, T.M.
Kam-Morgan, L.
Wang, A.
Eisenberg, M.
Rowland, C.M.
Hessling, J.J.
Broder, S.E.
Sninsky, J.J.
Tutt, A.
Anderson, S.
Chang, S.-.
(2009). A Single-Tube Quantitative Assay for mRNA Levels of Hormonal and Growth Factor Receptors in Breast Cancer Specimens. Journal of molecular diagnostics,
Vol.11
(2),
pp. 117-130.
Loi, S.
Haibe-Kains, B.
Lallemand, F.
Pusztai, L.
Tutt, A.
Gillett, C.E.
Piccart, M.
Phillips, W.A.
McArthur, G.
Sotiriou, C.
(2009). PIK3CA, AKT1 MUTATION AND HER2 AMPLIFICATION GENE SIGNATURES (GS) SUGGEST PREDOMINANTLY NEGATIVE FEEDBACK INHIBITION OF PI3K/AKT PATHWAY IN HUMAN BREAST CANCER (BC). Annals of oncology,
Vol.20,
pp. 45-45.
Rhiem, K.
Wappenschmidt, B.
Bosse, K.
Koeppler, H.
Tutt, A.N.
Schmutzler, R.K.
(2009). Platinum Sensitivity in a BRCA1 Mutation Carrier with Advanced Breast Cancer. Clinical oncology,
Vol.21
(6),
pp. 448-450.
Fong, P.C.
Boss, D.S.
Yap, T.A.
Tutt, A.
Wu, P.
Mergui-Roelvink, M.
Mortimer, P.
Swaisland, H.
Lau, A.
O'Connor, M.J.
Ashworth, A.
Carmichael, J.
Kaye, S.B.
Schellens, J.H.
de Bono, J.S.
(2009). Inhibition of poly(ADP-ribose) polymerase in tumors from BRCA mutation carriers. N engl j med,
Vol.361
(2),
pp. 123-134.
show abstract
BACKGROUND: The inhibition of poly(adenosine diphosphate [ADP]-ribose) polymerase (PARP) is a potential synthetic lethal therapeutic strategy for the treatment of cancers with specific DNA-repair defects, including those arising in carriers of a BRCA1 or BRCA2 mutation. We conducted a clinical evaluation in humans of olaparib (AZD2281), a novel, potent, orally active PARP inhibitor. METHODS: This was a phase 1 trial that included the analysis of pharmacokinetic and pharmacodynamic characteristics of olaparib. Selection was aimed at having a study population enriched in carriers of a BRCA1 or BRCA2 mutation. RESULTS: We enrolled and treated 60 patients; 22 were carriers of a BRCA1 or BRCA2 mutation and 1 had a strong family history of BRCA-associated cancer but declined to undergo mutational testing. The olaparib dose and schedule were increased from 10 mg daily for 2 of every 3 weeks to 600 mg twice daily continuously. Reversible dose-limiting toxicity was seen in one of eight patients receiving 400 mg twice daily (grade 3 mood alteration and fatigue) and two of five patients receiving 600 mg twice daily (grade 4 thrombocytopenia and grade 3 somnolence). This led us to enroll another cohort, consisting only of carriers of a BRCA1 or BRCA2 mutation, to receive olaparib at a dose of 200 mg twice daily. Other adverse effects included mild gastrointestinal symptoms. There was no obvious increase in adverse effects seen in the mutation carriers. Pharmacokinetic data indicated rapid absorption and elimination; pharmacodynamic studies confirmed PARP inhibition in surrogate samples (of peripheral-blood mononuclear cells and plucked eyebrow-hair follicles) and tumor tissue. Objective antitumor activity was reported only in mutation carriers, all of whom had ovarian, breast, or prostate cancer and had received multiple treatment regimens. CONCLUSIONS: Olaparib has few of the adverse effects of conventional chemotherapy, inhibits PARP, and has antitumor activity in cancer associated with the BRCA1 or BRCA2 mutation. (ClinicalTrials.gov number, NCT00516373.).
Natrajan, R.
Lambros, M.B.
Rodríguez-Pinilla, S.M.
Moreno-Bueno, G.
Tan, D.S.
Marchió, C.
Vatcheva, R.
Rayter, S.
Mahler-Araujo, B.
Fulford, L.G.
Hungermann, D.
Mackay, A.
Grigoriadis, A.
Fenwick, K.
Tamber, N.
Hardisson, D.
Tutt, A.
Palacios, J.
Lord, C.J.
Buerger, H.
Ashworth, A.
Reis-Filho, J.S.
(2009). Tiling path genomic profiling of grade 3 invasive ductal breast cancers. Clin cancer res,
Vol.15
(8),
pp. 2711-2722.
show abstract
PURPOSE: To characterize the molecular genetic profiles of grade 3 invasive ductal carcinomas of no special type using high-resolution microarray-based comparative genomic hybridization (aCGH) and to identify recurrent amplicons harboring putative therapeutic targets associated with luminal, HER-2, and basal-like tumor phenotypes. EXPERIMENTAL DESIGN: Ninety-five grade 3 invasive ductal carcinomas of no special type were classified into luminal, HER-2, and basal-like subgroups using a previously validated immunohistochemical panel. Tumor samples were microdissected and subjected to aCGH using a tiling path 32K BAC array platform. Selected regions of recurrent amplification were validated by means of in situ hybridization. Expression of genes pertaining to selected amplicons was investigated using quantitative real-time PCR and gene silencing was done using previously validated short hairpin RNA constructs. RESULTS: We show that basal-like and HER-2 tumors are characterized by "sawtooth" and "firestorm" genetic patterns, respectively, whereas luminal cancers were more heterogeneous. Apart from confirming known amplifications associated with basal-like (1q21, 10p, and 12p), luminal (8p12, 11q13, and 11q14), and HER-2 (17q12) cancers, we identified previously unreported recurrent amplifications associated with each molecular subgroup: 19q12 in basal-like, 1q32.1 in luminal, and 14q12 in HER-2 cancers. PPM1D gene amplification (17q23.2) was found in 20% and 8% of HER-2 and luminal cancers, respectively. Silencing of PPM1D by short hairpin RNA resulted in selective loss of viability in tumor cell lines harboring the 17q23.2 amplification. CONCLUSIONS: Our results show the power of aCGH analysis in unraveling the genetic profiles of specific subgroups of cancer and for the identification of novel therapeutic targets..
Tutt, A.
Robson, M.
Garber, J.E.
Domchek, S.
Audeh, M.W.
Weitzel, J.N.
Friedlander, M.
Carmichael, J.
(2009). Phase II trial of the oral PARP inhibitor olaparib in BRCA-deficient advanced breast cancer. J clin oncol,
Vol.27
(18_suppl),
p. CRA501.
show abstract
CRA501 Background: Olaparib (AZD2281; KU-0059436) is a novel, orally active PARP inhibitor that induces synthetic lethality in homozygous BRCA-deficient cells. A phase I trial identified 400 mg bd as the maximum tolerated dose (MTD) with an initial signal of efficacy in BRCA-deficient ovarian cancers (ASCO 2008; abst 5510). The primary aim of this study was to test the efficacy of olaparib in confirmed BRCA1/BRCA2 carriers with advanced refractory breast cancer. The secondary aim was to assess safety and tolerability in this population. METHODS: In an international, multicenter, proof-of-concept, single-arm, phase II study, two sequential patient (pt) cohorts received continuous oral olaparib in 28-day cycles initially at the MTD, 400 mg bd (27 pts), and subsequently at 100 mg bd, a previously identified PARP inhibitory dose (27 pts). Eligibility criteria included confirmed BRCA1/BRCA2 mutation and recurrent, measurable chemotherapy-refractory breast cancer. The primary efficacy endpoint was best objective response rate (ORR; RECIST) post baseline. Progression-free survival (PFS) and clinical benefit rate were secondary endpoints. All adverse events were reported using CTCAE v3. RESULTS: On November 20, 2008, 54 pts exposed to a median of three prior lines of chemotherapy had been enrolled. 27 pts were dosed at 400 mg bd (18 BRCA1 deficient and 9 BRCA2 deficient), and 24 of these had databased RECIST assessments. The ORR (currently based on unconfirmed responses) was 38% (9/24) (400 mg bd). Causally-related toxicity was mainly mild (grade 1-2) in severity; 9/27 pts (33%) had fatigue; 7/27 (26%) had nausea; 4/27 (15%) had vomiting; and 1/27 (4%) had anemia. Causally-related grade 3 or higher toxicities were seen in 5 pts (19%) with fatigue (3 pts), nausea (2 pts), and anemia (1 pt). 27 pts were treated in the subsequent 100 mg bd cohort where no data are currently available. CONCLUSIONS: Olaparib at 400 mg bd is well tolerated and highly active in advanced chemotherapy-refractory BRCA-deficient breast cancer. Toxicity in BRCA1/BRCA2 carriers was similar to that reported previously in non-carriers. This first study with olaparib in BRCA-deficient breast cancers provides positive proof of concept for high activity and tolerability of a genetically defined targeted therapy. [Table: see text]..
Tutt, A.
Robson, M.
Garber, J.E.
Domchek, S.
Audeh, M.W.
Weitzel, J.N.
Friedlander, M.
Carmichael, J.
(2009). Phase II trial of the oral PARP inhibitor olaparib in BRCA-deficient advanced breast cancer. J clin oncol,
Vol.27
(15_suppl),
p. CRA501.
show abstract
CRA501 The full, final text of this abstract will be available in Part II of the 2009 ASCO Annual Meeting Proceedings, distributed onsite at the Meeting on May 30, 2009, and as a supplement to the June 20, 2009, issue of the Journal of Clinical Oncology. [Table: see text]..
Tutt, A.
Ashworth, A.
(2008). Can genetic testing guide treatment in breast cancer?. European journal of cancer,
Vol.44
(18),
pp. 2774-7.
Reis-Filho, J.S.
Tutt, A.N.
(2008). Triple negative tumours: a critical review. Histopathology,
Vol.52
(1),
pp. 108-118.
show abstract
Breast cancer is a heterogeneous disease that encompasses several distinct entities with remarkably different biological characteristics and clinical behaviour. Currently, breast cancer patients are managed according to algorithms based on a constellation of clinical and histopathological parameters in conjunction with assessment of hormone receptor (oestrogen and progesterone receptor) status and HER2 overexpression/gene amplification. Although effective tailored therapies have been developed for patients with hormone receptor-positive or HER2+ disease, chemotherapy is the only modality of systemic therapy for patients with breast cancers lacking the expression of these markers (triple-negative cancers). Recent microarray expression profiling analyses have demonstrated that breast cancers can be systematically characterized into biologically and clinically meaningful groups. These studies have led to the re-discovery of basal-like breast cancers, which preferentially show a triple-negative phenotype. Both triple-negative and basal-like cancers preferentially affect young and African-American women, are of high histological grade and have more aggressive clinical behaviour. Furthermore, a significant overlap between the biological and clinical characteristics of sporadic triple-negative and basal-like cancers and breast carcinomas arising in BRCA1 mutation carriers has been repeatedly demonstrated. In this review, we critically address the characteristics of basal-like and triple-negative cancers, their similarities and differences, their response to chemotherapy as well as strategies for the development of novel therapeutic targets for these aggressive types of breast cancer. In addition, the possible mechanisms are discussed leading to BRCA1 pathway dysfunction in sporadic triple-negative and basal-like cancers and animal models for these tumour types..
Iorns, E.
Turner, N.C.
Elliott, R.
Syed, N.
Garrone, O.
Gasco, M.
Tutt, A.N.
Crook, T.
Lord, C.J.
Ashworth, A.
(2008). Identification of CDK10 as an important determinant of resistance to endocrine therapy for breast cancer. Cancer cell,
Vol.13
(2),
pp. 91-104.
show abstract
Therapies that target estrogen signaling have transformed the treatment of breast cancer. However, the effectiveness of these agents is limited by the development of resistance. Here, an RNAi screen was used to identify modifiers of tamoxifen sensitivity. We demonstrate that CDK10 is an important determinant of resistance to endocrine therapies and show that CDK10 silencing increases ETS2-driven transcription of c-RAF, resulting in MAPK pathway activation and loss of tumor cell reliance upon estrogen signaling. Patients with ER alpha-positive tumors that express low levels of CDK10 relapse early on tamoxifen, demonstrating the clinical significance of these observations. The association of low levels of CDK10 with methylation of the CDK10 promoter suggests a mechanism by which CDK10 expression is reduced in tumors..
Turner, N.C.
Lord, C.J.
Iorns, E.
Brough, R.
Swift, S.
Elliott, R.
Rayter, S.
Tutt, A.N.
Ashworth, A.
(2008). A synthetic lethal siRNA screen identifying genes mediating sensitivity to a PARP inhibitor. Embo journal,
Vol.27
(9),
pp. 1368-10.
Tutt, A.
Wang, A.
Rowland, C.
Gillett, C.
Lau, K.
Chew, K.
Dai, H.
Kwok, S.
Ryder, K.
Shu, H.
Springall, R.
Cane, P.
McCallie, B.
Kam-Morgan, L.
Anderson, S.
Buerger, H.
Gray, J.
Bennington, J.
Esserman, L.
Hastie, T.
Broder, S.
Sninsky, J.
Brandt, B.
Waldman, F.
(2008). Risk estimation of distant metastasis in node-negative, estrogen receptor-positive breast cancer patients using an RT-PCR based prognostic expression signature. Bmc cancer,
Vol.8.
Hiley, C.
Tutt, A.
Torres, M.
Palmieri, C.
(2008). Adjuvant radiotherapy for breast cancer. British medical journal,
Vol.337.
Turner, N.C.
Reis-Filho, J.S.
Russell, A.M.
Springall, R.J.
Ryder, K.
Steele, D.
Savage, K.
Gillett, C.E.
Schmitt, F.C.
Ashworth, A.
Tutt, A.N.
(2007). BRCA1 dysfunction in sporadic basal-like breast cancer. Oncogene,
Vol.26
(14),
pp. 2126-2132.
show abstract
Basal-like breast cancers form a distinct subtype of breast cancer characterized by the expression of markers expressed in normal basal/myoepithelial cells. Breast cancers arising in carriers of germline BRCA1 mutations are predominately of basal-like type, suggesting that BRCA1 dysfunction may play a role in the pathogenesis of sporadic basal-like cancers. We analysed 37 sporadic breast cancers expressing the basal marker cytokeratin 5/6, and age- and grade-matched controls, for downregulation of BRCA1. Although BRCA1 promoter methylation was no more common in basal-like cancers (basal 14% vs controls 11%, P=0.72), BRCA1 messenger RNA expression was twofold lower in basal-like breast cancers compared to matched controls (P=0.008). ID4, a negative regulator of BRCA1, was expressed at 9.1-fold higher levels in basal-like breast cancer (P<0.0001), suggesting a potential mechanism of BRCA1 downregulation. BRCA1 downregulation correlated with the presence of multiple basal markers, revealing heterogeneity in the basal-like phenotype. Finally, we found that 63% of metaplastic breast cancers, a rare type of basal-like cancers, had BRCA1 methylation, in comparison to 12% of controls (P<0.0001). The high prevalence of BRCA1 dysfunction identified in this study could be exploited in the development of novel approaches to targeted treatment of basal-like breast cancer..
Rakha, E.A.
Tan, D.S.
Foulkes, W.D.
Ellis, I.O.
Tutt, A.
Nielsen, T.O.
Reis-Filho, J.S.
(2007). Are triple-negative tumours and basal-like breast cancer synonymous?. Breast cancer res,
Vol.9
(6),
p. 404.
Shanley, S.
McReynolds, K.
Ardern-Jones, A.
Ahern, R.
Fernando, I.
Yarnold, J.
Evans, G.
Eccles, D.
Hodgson, S.
Ashley, S.
Ashcroft, L.
Tutt, A.
Bancroft, E.
Short, S.
Gui, G.
Breast Unit of the Royal Marsden NHS Foundation Trust,
Barr, L.
Baildam, A.
Howell, A.
Royle, G.
Pierce, L.
Easton, D.
Eeles, R.
(2006). Late toxicity is not increased in BRCA1/BRCA2 mutation carriers undergoing breast radiotherapy in the United Kingdom. Clin cancer res,
Vol.12
(23),
pp. 7025-7032.
show abstract
PURPOSE: To undertake the first substantial clinical study of breast radiotherapy toxicity in BRCA1 and BRCA2 mutation carriers in the United Kingdom. EXPERIMENTAL DESIGN: Acute and late radiation effects were evaluated in a retrospective study of 55 BRCA1 and BRCA2 mutation carriers treated with radiotherapy for breast cancer at four centers between 1983 and 2002. Individual matching with controls who had sporadic breast cancer was undertaken for age at diagnosis, time since completion of radiation, and treatment variables. Detailed assessments were undertaken by one examiner. Median follow-up was 6.75 years for carriers and 7.75 years for controls. Rates of late events (rib fractures, lung fibrosis, necrosis of soft tissue/bone, and pericarditis) as well as LENT-SOMA scores and clinical photography scores of breast size, shape, and skin telangiectasia were the primary end points. RESULTS: No increase in clinically significant late toxicity was seen in the mutation carriers. CONCLUSIONS: These data add substantial weight to the evidence that the outcomes in the treated breast from radiotherapy in women with BRCA1 or BRCA2 mutations are comparable with those in women with sporadic breast cancer..
Shanley, S.
McReynolds, K.
Ardern-Jones, A.
Ahern, R.
Fernando, I.
Yarnold, J.
Evans, G.
Eccles, D.
Hodgson, S.
Ashley, S.
Ashcroft, L.
Tutt, A.
Bancroft, E.
Short, S.
Smith, I.
Gui, G.
Royal Marsden NHS Foundation Trust,
Barr, L.
Baildam, A.
Howell, A.
Royle, G.
Pierce, L.
Easton, D.
Eeles, R.
(2006). Acute chemotherapy-related toxicity is not increased in BRCA1 and BRCA2 mutation carriers treated for breast cancer in the United Kingdom. Clin cancer res,
Vol.12
(23),
pp. 7033-7038.
show abstract
PURPOSE: To evaluate acute toxicity induced by chemotherapy for breast cancer in a retrospective study of 62 BRCA1/2 mutation carriers matched 1:1 with women who had treatment for sporadic disease in the United Kingdom between 1983 and 2003. EXPERIMENTAL DESIGN: All participants were interviewed by one of two researchers using standardized questionnaires, and their medical records were reviewed by one research nurse. The two main regimens received were cyclophosphamide, methotrexate, and fluorouracil and fluorouracil, epirubicin, and cyclophosphamide. The proportion of cases and controls receiving anthracycline-based treatment was equivalent, but fewer BRCA1 cases received this treatment than did BRCA2 mutation carriers. Toxicity was documented using the Eastern Cooperative Oncology Group Common Toxicity Criteria for hematologic, infective, and gastrointestinal toxicities. No increase in toxicity was seen in BRCA1/2 mutation carriers. RESULTS: The only significant difference was that neutropenia was less evident in BRCA2 mutation carriers than in either BRCA1 mutation carriers or controls. As a result, there was no requirement for dose reduction among BRCA2 mutation carriers, in contrast to 10 of 39 BRCA1 carriers and 16 of 62 controls (P = 0.02). CONCLUSIONS: This result has implications for therapy and indicates that women with mutations in BRCA1 and BRCA2 may be given the same doses of chemotherapy as noncarriers..
Tutt, A.
Yarnold, J.
(2006). Radiobiology of breast cancer. Clin oncol (r coll radiol),
Vol.18
(3),
pp. 166-178.
show abstract
Advances in molecular and cellular biology are transforming our understanding of breast cancer and promise the same for radiotherapy over the next few years. At the clinical level, the molecular basis of fractionation dependency and other tumour and normal tissue responses are likely to become clearer. More importantly, they will become useful in the clinic, where molecular characterisation of the patient and tumour will start to determine therapeutic options. Although many of the fundamental processes are only amenable to study in laboratory systems, the power of array-based technologies makes it possibly to address highly relevant questions in the clinic, using functional imaging and/or tissue biopsies. To help clinical oncologists exploit these opportunities in translational research, some aspects of the molecular and cellular basis of radiotherapy are described below in their relation to breast cancer..
de Bono, J.S.
Fong, P.C.
Boss, D.
Spicer, J.
Roelvink, M.
Tutt, A.
Mortimer, P.
O'Connor, M.
Schellens, J.H.
Kaye, S.B.
(2006). Phase I pharmacolkinetic (PK) and pharmacodynamic (PD) evaluation of an oral small molecule inhibitor of Poly ADP-Ribose Polymerase (PARP), Ku in patients (p) with advanced tumours. Ejc supplements,
Vol.4
(12),
pp. 153-153.
McCabe, N.
Turner, N.C.
Lord, C.J.
Kluzek, K.
Bialkowska, A.
Swift, S.
Giavara, S.
O'Connor, M.J.
Tutt, A.N.
Zdzienicka, M.Z.
Smith, G.C.
Ashworth, A.
(2006). Deficiency in the repair of DNA damage by homologous recombination and sensitivity to poly(ADP-ribose) polymerase inhibition. Cancer res,
Vol.66
(16),
pp. 8109-8115.
show abstract
Deficiency in either of the breast cancer susceptibility proteins BRCA1 or BRCA2 induces profound cellular sensitivity to the inhibition of poly(ADP-ribose) polymerase (PARP) activity. We hypothesized that the critical role of BRCA1 and BRCA2 in the repair of double-strand breaks by homologous recombination (HR) was the underlying reason for this sensitivity. Here, we examine the effects of deficiency of several proteins involved in HR on sensitivity to PARP inhibition. We show that deficiency of RAD51, RAD54, DSS1, RPA1, NBS1, ATR, ATM, CHK1, CHK2, FANCD2, FANCA, or FANCC induces such sensitivity. This suggests that BRCA-deficient cells are, at least in part, sensitive to PARP inhibition because of HR deficiency. These results indicate that PARP inhibition might be a useful therapeutic strategy not only for the treatment of BRCA mutation-associated tumors but also for the treatment of a wider range of tumors bearing a variety of deficiencies in the HR pathway or displaying properties of 'BRCAness.'.
Fong, P.C.
Spicer, J.
Reade, S.
Reid, A.
Vidal, L.
Schellens, J.H.
Tutt, A.
Harris, P.A.
Kaye, S.
De Bono, J.S.
(2006). Phase I pharmacokinetic (PK) and pharmacodynamic (PD) evaluation of a small molecule inhibitor of Poly ADP-Ribose Polymerase (PARP), KU-0059436 (Ku) in patients (p) with advanced tumours. J clin oncol,
Vol.24
(18_suppl),
p. 3022.
show abstract
3022 Background: PARP is a DNA strand break and base damage repair enzyme. Ku inhibits PARP-1 and 2 with a mean IC50 of 2nM. Inhibition of PARP leads to defective DNA repair and induces selective cytotoxicity in cells with defective homologous recombination repair through, for example, loss of BRCA 1/2 function (Farmer et al, Nature 2005 Apr 14;434(7035):917-21). This is a first in man Phase I trial of Ku. PD studies included functional evaluation of PARP-1 activity in surrogate and tumor tissue. METHODS: Ku was administered once a day for 14 of every 21 days to p with advanced solid tumors refractory to standard treatment. Cohorts of 3-6 p were treated, with a starting dose of 10 mg/day. The dose was doubled in the absence of drug related grade 2 CTC-AE toxicity. Drug related toxicity in cancer patients known to carry a BRCA mutation is being compared to toxicity in other patients. RESULTS: To date 12 p (6 male; median age 55y, range 25-73) with solid tumours have received 25 courses (range 1-8), with 1 p being a known BRCA1 mutation carrier (dose level 40mg). Dose levels evaluated to date include 10, 20, 40, and 80mg per day. Minimal toxicity has been observed to date, with only intermittent grade 1 nausea being reported. PK data to date, up to 40mg per day, supports dose proportionality with a mean elimination half-life of 6.91 hours (Range: 5.3-9.5), a mean clearance of 4.1 L/h (Range: 1.3-9.4) and a mean volume of distribution of 39.8 L (Range: 16.3-86.9). PD studies indicate inhibition of PARP functional activity in peripheral blood mononuclear cells with increasing inhibition observed with increasing dose of Ku. Initial studies in tumor biopsies performed pre-treatment and on day 8 revealed PARP inhibition of >50% at 40 mg/day. A p with metastatic soft tissue sarcoma and progressing disease pretreatment achieved stable disease for 24 weeks. CONCLUSIONS: Dose escalation continues with more BRCA carriers planned. PARP inhibition in both surrogate and tumor tissue is achievable with minimal toxicity in cancer patients, and has not resulted in any short term toxicity in a BRCA mutation carrier. [Table: see text]..
Turner, N.
Tutt, A.
Ashworth, A.
(2005). Targeting the DNA repair defect of BRCA tumours. Curr opin pharmacol,
Vol.5
(4),
pp. 388-393.
show abstract
Carriers of heterozygous mutations in BRCA1 or BRCA2 are strongly predisposed to breast and ovarian cancers. Cancers arising in these individuals have consistently lost the wild-type allele during tumour progression, and are therefore deficient in BRCA1 or BRCA2 function. Both BRCA1 and BRCA2 proteins have been implicated in the repair of double-strand DNA breaks by homologous recombination. This functional role in DNA repair could be exploited in the treatment of BRCA-deficient cancers by targeting the tumours with drugs that create DNA damage highly reliant on BRCA1 or BRCA2 for repair..
McCabe, N.
Lord, C.J.
Tutt, A.N.
Martin, N.M.
Smith, G.C.
Ashworth, A.
(2005). BRCA2-deficient CAPAN-1 cells are extremely sensitive to the inhibition of Poly (ADP-Ribose) polymerase: an issue of potency. Cancer biol ther,
Vol.4
(9),
pp. 934-936.
show abstract
We have previously demonstrated that deficiency of either the BRCA1 or BRCA2 breast cancer susceptibility proteins confers substantial cellular sensitivity to the inhibition of Poly(ADP-Ribose) polymerase (PARP). PARP is a key enzyme in the repair of single strand DNA damage via the Base Excision Repair pathway. We suggested that PARP inhibition produces persistent single-strand DNA breaks or gaps which degenerate into stalled replication forks and double-strand breaks, which may be repaired by homologous recombination, a process partially dependent on BRCA1 and BRCA2. It has recently been suggested that our results might be limited to certain BRCA2 mutations as the CAPAN-1 cell line, which carries a naturally occurring 6174delT mutation in one BRCA2 allele accompanied by loss of the wild-type allele, is apparently insensitive to two PARP inhibitors 3-aminobenzamide (IC50 33 microM) and NU1025 (IC50 400 nM). Here we show that CAPAN-1 cells are in fact very sensitive to the potent PARP inhibitors KU0058684 (IC50 3.2 nM) and KU0058948 (IC50 3.4 nM). In contrast, our results reveal much less sensitivity to a chemically related but much less active compound KU0051529 (IC50 730 nM) and to NU1025. These results confirm that treatment with potent PARP inhibitors remains an exciting potential therapy for cancers involving BRCA1 or BRCA2 deficiency..
Farmer, H.
McCabe, N.
Lord, C.J.
Tutt, A.N.
Johnson, D.A.
Richardson, T.B.
Santarosa, M.
Dillon, K.J.
Hickson, I.
Knights, C.
Martin, N.M.
Jackson, S.P.
Smith, G.C.
Ashworth, A.
(2005). Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature,
Vol.434
(7035),
pp. 917-921.
show abstract
BRCA1 and BRCA2 are important for DNA double-strand break repair by homologous recombination, and mutations in these genes predispose to breast and other cancers. Poly(ADP-ribose) polymerase (PARP) is an enzyme involved in base excision repair, a key pathway in the repair of DNA single-strand breaks. We show here that BRCA1 or BRCA2 dysfunction unexpectedly and profoundly sensitizes cells to the inhibition of PARP enzymatic activity, resulting in chromosomal instability, cell cycle arrest and subsequent apoptosis. This seems to be because the inhibition of PARP leads to the persistence of DNA lesions normally repaired by homologous recombination. These results illustrate how different pathways cooperate to repair damage, and suggest that the targeted inhibition of particular DNA repair pathways may allow the design of specific and less toxic therapies for cancer..
Turner, N.
Tutt, A.
Ashworth, A.
(2004). Hallmarks of 'BRCAness' in sporadic cancers. Nat rev cancer,
Vol.4
(10),
pp. 814-819.
show abstract
Germline mutations in the BRCA1, BRCA2 and Fanconi anaemia genes confer cancer susceptibility, and the proteins encoded by these genes have distinct functions in related DNA-repair processes. Emerging evidence indicates that these processes are disrupted by numerous mechanisms in sporadic cancers. Collectively, there are properties that define 'BRCAness' - that is, traits that some sporadic cancers share with those occurring in either BRCA1- or BRCA2-mutation carriers. These common properties might have important implications for the clinical management of these cancers..
Gudmundsdottir, K.
Lord, C.J.
Witt, E.
Tutt, A.N.
Ashworth, A.
(2004). DSS1 is required for RAD51 focus formation and genomic stability in mammalian cells. Embo rep,
Vol.5
(10),
pp. 989-993.
show abstract
full text
BRCA2 is a breast cancer susceptibility gene implicated in the repair of double-strand breaks by homologous recombination with RAD51. BRCA2 associates with a 70-amino-acid protein, DSS1, but the functional significance of this interaction has remained unclear. Recently, deficiency of a DSS1 orthologue in the fungus Ustilago maydis has been shown to cause a defect in recombinational DNA repair. Here we have investigated the consequences of DSS1 depletion in mammalian cells. We show that like BRCA2, DSS1 is required for DNA damage-induced RAD51 focus formation and for the maintenance of genomic stability, indicating a function conserved from lower eukaryotes to humans. However, DSS1 seems to be not required for BRCA2 or RAD51 stability or for BRCA2 and RAD51 to interact, raising the possibility that DSS1 may be required for the BRCA2-RAD51 complex to become associated with sites of DNA damage..
Tutt, A.
Connor, F.
Bertwistle, D.
Kerr, P.
Peacock, J.
Ross, G.
Ashworth, A.
(2003). Cell cycle and genetic background dependence of the effect of loss of BRCA2 on ionizing radiation sensitivity. Oncogene,
Vol.22
(19),
pp. 2926-2931.
show abstract
Carriers of mutations in the BRCA2 gene are at a highly elevated risk of breast and other cancers. The BRCA2 gene encodes a very large protein thought to play a role in DNA repair. To examine the effect of mutation of BRCA2 on sensitivity to ionizing radiation, we used a previously described mouse model system (Brca2(Tr)) in which the Brca2 open reading frame is truncated. Mouse embryo fibroblasts carrying this mutation have a proliferative defect, which we show here can be substantially rescued by genetic ablation of p53. Proliferating Brca2(Tr/Tr)/p53(-/-) cells, like Brca2(Tr/Tr) cells, show genomic instability. We used the clonogenic survival assay, which depends on the ability of cells to proliferate, to examine the cell cycle dependence of radiation sensitivity of Brca2(Tr/Tr)/p53(-/-) compared to p53(-/-) and wild-type cells. This showed that the Brca2 mutation had little effect on cells irradiated in quiescence but sensitized proliferating cells to ionizing radiation on a p53(-/-) background. These results suggest that the major role of Brca2 in mediating cell survival after irradiation is in the S and G(2) phases of the cell cycle..
Tutt, A.N.
van Oostrom, C.T.
Ross, G.M.
van Steeg, H.
Ashworth, A.
(2002). Disruption of Brca2 increases the spontaneous mutation rate in vivo: synergism with ionizing radiation. Embo rep,
Vol.3
(3),
pp. 255-260.
show abstract
full text
The breast cancer predisposition gene BRCA2 encodes a protein involved in the repair of DNA double-strand breaks, which arise spontaneously and following exposure to ionizing radiation (IR). To develop a mouse model that examines the effect of BRCA2 mutation and IR exposure on in vivo somatic mutation acquisition, we crossed mice with targeted disruption of Brca2 with a LacZ transgenic mutation reporter strain. Loss of both wild-type Brca2 alleles caused a 2.3-fold increase, equivalent to an extra 100 mutations per cell, in the in vivo acquisition of spontaneous somatic mutation by 2 weeks gestation. IR (4 Gy) had a disproportionate effect on animals homozygous for Brca2 disruption, inducing 3.4-fold more mutations compared with wild-type animals. These data provide the first evidence that loss of Brca2 increases in vivo somatic mutation acquisition and synergizes with IR exposure, with potential attendant implications for mammographic screening and therapeutic IR in mutation carriers..
Tutt, A.
Ashworth, A.
(2002). The relationship between the roles of BRCA genes in DNA repair and cancer predisposition. Trends mol med,
Vol.8
(12),
pp. 571-576.
show abstract
The proteins encoded by the breast-cancer-susceptibility genes, BRCA1 and BRCA2, have recently been implicated in DNA-repair processes, thereby improving our understanding of how the loss of these genes contributes to cancer initiation and progression. It appears that the role of BRCA1 in DNA repair, which could involve the integration of several pathways, is broader than that of BRCA2. BRCA1 functions in the signalling of DNA damage and its repair by homologous recombination, nucleotide-excision repair and possibly non-homologous end-joining. BRCA2 has a more specific role in DNA repair, regulating the activity of RAD51, which is required for homologous recombination. An improved understanding of the interactions of BRCA1 and BRCA2 with other proteins in large macromolecular complexes is helping to reveal their exact role in DNA repair..
Tutt, A.
Bertwistle, D.
Valentine, J.
Gabriel, A.
Swift, S.
Ross, G.
Griffin, C.
Thacker, J.
Ashworth, A.
(2001). Mutation in Brca2 stimulates error-prone homology-directed repair of DNA double-strand breaks occurring between repeated sequences. Embo j,
Vol.20
(17),
pp. 4704-4716.
show abstract
full text
Mutation of BRCA2 causes familial early onset breast and ovarian cancer. BRCA2 has been suggested to be important for the maintenance of genome integrity and to have a role in DNA repair by homology- directed double-strand break (DSB) repair. By studying the repair of a specific induced chromosomal DSB we show that loss of Brca2 leads to a substantial increase in error-prone repair by homology-directed single-strand annealing and a reduction in DSB repair by conservative gene conversion. These data demonstrate that loss of Brca2 causes misrepair of chromosomal DSBs occurring between repeated sequences by stimulating use of an error-prone homologous recombination pathway. Furthermore, loss of Brca2 causes a large increase in genome-wide error-prone repair of both spontaneous DNA damage and mitomycin C-induced DNA cross-links at the expense of error-free repair by sister chromatid recombination. This provides insight into the mechanisms that induce genome instability in tumour cells lacking BRCA2..
Tutt, A.N.
Bertwistle, D.
Valentine, J.
Gabriel, A.
Swift, S.
Ross, G.M.
Griffin, C.
Thacker, J.
Ashworth, A.
(2001). Mutation in BRCA2 reduces use of error-free DNA repair by sister chromatid recombination and stimulates error-prone DNA repair by homology directed single-strand annealing. Breast cancer research and treatment,
Vol.69
(3),
pp. 255-255.
Tutt, A.
Gabriel, A.
Bertwistle, D.
Connor, F.
Paterson, H.
Peacock, J.
Ross, G.
Ashworth, A.
(1999). Absence of Brca2 causes genome instability by chromosome breakage and loss associated with centrosome amplification. Curr biol,
Vol.9
(19),
pp. 1107-1110.
show abstract
Women heterozygous for mutations in the breast-cancer susceptibility genes BRCA1 and BRCA2 have a highly elevated risk of developing breast cancer [1]. BRCA1 and BRCA2 encode large proteins with no sequence similarity to one another. Although involvement in DNA repair and transcription has been suggested, it is still not understood how loss of function of these genes leads to breast cancer [2]. Embryonic fibroblasts (MEFs) derived from mice homozygous for a hypomorphic mutation (Brca2(Tr2014)) within the 3' region of exon 11 in Brca2 [3], or a similar mutation (Brca2(Tr)) [4], proliferate poorly in culture and overexpress the tumour suppressor p53 and the cyclin-dependent kinase inhibitor p21(Waf1/Cip1). These MEFs have intact p53-dependent DNA damage G(1)-S [3] [4] and G(2)-M checkpoints [4], but are impaired in DNA double-strand break repair [3] and develop chromosome aberrations [4]. Here, we report that Brca2(Tr2014/Tr2014) MEFs frequently develop micronuclei. These abnormal DNA-containing bodies were formed through both loss of acentric chromosome fragments and by chromosome missegregation, which resulted in aneuploidy. Absence of Brca2 also led to centrosome amplification, which we found associated with the formation of micronuclei. These data suggest a potential mechanism whereby loss of BRCA2 may, within subclones, drive the loss of cell-cycle regulation genes, enabling proliferation and tumourigenesis..
Tutt, A.N.
A'hern, R.
Henk, J.M.
(1998). Cervical node metastasis of unknown primary site: The Royal Marsden Hospital 15 year experience. British journal of cancer,
Vol.78,
pp. 6-6.
de Rinaldis, E.
Gazinska, P.
Mera, A.
Modrusan, Z.
Fedorowicz, G.M.
Burford, B.
Gillett, C.
Marra, P.
Grigoriadis, A.
Dornan, D.
Holmberg, L.
Pinder, S.
Tutt, A.
Integrated genomic analysis of triple-negative breast cancers reveals novel microRNAs associated with clinical and molecular phenotypes and sheds light on the pathways they control. Bmc genomics,
(14).