Pathogenic BRCA1 Mutations Research Articles

Patients with BRCA mutations have superior outcomes after intraperitoneal chemotherapy in optimally resected high grade ovarian cancer

ObjectivesTo compare the outcomes after intraperitoneal (IP) chemotherapy in patients with and without pathogenic BRCA mutations. MethodsPatients with high grade ovarian cancer who were treated with adjuvant IP chemotherapy in the initial setting between 2005 and 2016 were identified. Outcomes were compared between patients with pathogenic mutations in BRCA (BRCA+) and those who tested negative or were unknown (BRCA−). ResultsA total of 100 eligible patients were identified. The median follow-up was 47.0 months (range, 6.6–144.1 months). Of these 100 patients, 77 patients underwent BRCA testing; 25 patients (32%) were BRCA+ (23 germline, 2 somatic). No differences were noted between groups with respect to number of IP cycles, stage, or residual disease after surgery. The median progression-free survival (PFS) was longer in the BRCA+ group; median PFS was not reached in the BRCA+ group compared to 17.3 months in the BRCA− group (HR = 0.38; 95% CI 0.20–0.73, P = 0.003). Median overall survival (OS) was longer in the BRCA+ group at 110.4 months versus 67.1 months (HR = 0.28, 95% CI 0.11–0.73, P = 0.009). ConclusionsPathogenic BRCA mutations are more common than expected in optimally resected ovarian cancer patients selected for IP therapy. IP therapy was associated with a dramatic improvement in PFS and OS in BRCA+ patients compared with BRCA− patients. This improvement is greater than has been reported for BRCA+ patients with IV chemotherapy. The magnitude of this benefit suggests that patients with pathogenic mutations in BRCA may benefit from IP therapy.

Neurofeedback in Psychological Rehabilitation of Stroke Patients

Tissue genomic heterogeneity (t-HET) in patients with epithelial ovarian cancer (OVCA) is related to tissue plasticity, i.e., flexibility to adapt to adverse molecular environments. Here, we interrogated the presence and clinical relevance of OVCA t-HET.We applied high-depth (>2000×) sequencing on 297 paraffin tissue samples (fallopian tubes, ovaries, intra-abdominal metastases) from 71 treatment-naïve patients who subsequently received first-line platinum-based chemotherapy. Based on tissue mutation patterns, we distinguished tissue genotypes into: no mutation (33/297 samples; 11.1%), stable (173; 58.2%) and unstable (91; 30.7%). We profiled genotypes per patient and assessed t-HET in 69 patients. Predicted pathogenic mutations refer to germline and/or tissues.Among all 71 patients, 46 (64.8%) had pathogenic BRCA1 mutations and 15 (21.7%) had BRCA1/2 disruption (i.e., pathogenic mutations with position-LOH). We classified 29 patients with t-HET (42%), all with pathogenic BRCA1; t-HET was observed in 64% with such mutations (p < 0.001). As opposed to non-t-HET, matched tissues in t-HET shared pathogenic BRCA1 (p < 0.001) but not BRCA2 and TP53. Germline BRCA1 mutations in tissues exhibited position-LOH; heterozygous status; or, partial loss of the inherited allele accompanied by additional clonal mutations. Patients with t-HET had worse outcome (log-rank p = 0.048 [progression-free]; p = 0.037 [overall survival]), including 12/15 patients with disrupted BRCA1/2 and 3 BRCA1 carriers with partial germline loss in tissues.Pathogenic BRCA1 mutations appear necessary but may not be sufficient for the establishment of t-HET. t-HET may be associated with worse outcome, including in patients with disrupted BRCA1/2, which is usually considered as a favourable marker. OVCA t-HET may need to be addressed for treatment decisions.

Prevalence and Spectrum of Breast Cancer Inherited Mutations in Uganda and Cameroon Women

Background: Breast cancer among indigenous Africans is characterized by higher prevalence of triple-negative disease and poor prognosis. A previous study in Nigeria reported a strikingly high prevalence of deleterious germline mutations in BRCA1 and BRCA2 among Nigerian women with breast cancer. It is unknown if this is true in other sub-Saharan African countries. Aim: The objective of this study is to determine the frequency of germline mutations among an unselected sample of women in Africa. Methods: We conducted a case-control study of breast cancer in Uganda and Cameroon to investigate genetic and nongenetic risk factors for breast cancer. Breast cancer cases were enrolled in two tertiary hospitals in the two countries, unselected for age at diagnosis and family history. Controls who were free of breast cancer were enrolled in the same hospitals and matched to cases on age. A 24-gene sequencing panel was used to test germline mutations in cases and controls. Results: There were 176 cases and 177 controls with a mean age at diagnosis of 46.2 years for cases and mean age at interview of 46.7 years for controls. Among cases, 18.2% carried a pathogenic mutation in a breast cancer gene: 6.3% in BRCA1, 6.3% in BRCA2, 1.7% in ATM, 1.1% in PALB2, 0.6% in BARD1, 0.6% in CDH1, 0.6% in TP53, and 1.2% in any of 17 other genes. Among controls, 2.3% carried a pathogenic mutation in one of the 24 susceptibility genes. Cases were 9.6-fold more likely to carry a mutation compared with controls (odds ratio=9.61, 95% confidence interval: 3.28-38.1; P &lt; 0.001). The mean age of breast cancer cases with pathogenic BRCA1 mutations was 38.3 years compared with 46.7 years among other cases without such mutations ( P = 0.03). There was a trend that cases with a positive family history had higher chance of carrying a mutation (33.3%) than cases without (17.1%), but few cases reported a positive family history. Conclusion: Our findings confirm the earlier report of a high proportion of deleterious mutations in BRCA1 and BRCA2 among breast cancer patients in sub-Saharan Africa. As most of these women present with advanced breast cancer, there is an urgent need to improve access to genomic testing and life saving cancer medicines including chemotherapy and clinical trials of novel agents like PARP inhibitors. Given the high burden of inherited breast cancer, genetic risk assessment should be integrated into cancer control plans in sub-Saharan Africa.

Abstract B51: Genetic etiology of ER low/HER2- breast tumors

Abstract Background: In 2010, the American Society of Clinical Oncology (ASCO) and the College of American Pathologists (CAP) issued guidelines that tumors with &amp;gt;1% positively staining cells should be considered ER positive. Numerous studies have demonstrated that tumors with 1-9% ER-positive cells (low staining) represent ER-negative intrinsic subtypes with outcomes similar to ER-negative tumors. Given the similarities between low-staining tumors and TNBC, we investigated whether low-staining tumors share a germline etiology with TNBC. Methods: The Clinical Breast Care Project was queried to identify all tumors with percent ER-positive cells and HER2 data available. Genomic DNA was isolated from available blood samples from patients with HER2-negative tumors with either negative (TNBC) or low-staining (1-9%) ER status. Targeted sequencing was performed using the Illumina TruSight Cancer panel. Pathogenic mutations were identified using VariantStudio and classified as pathogenic, likely pathogenic, uncertain significance (VUS), likely benign, or benign using ClinVar. Results: Low ER/HER2- tumors and TNBC represented 50 (2.7%) and 223 (12.2%) of the 1819 tumors in the database. Patients with low-staining tumors and TNBC did not differ significantly by age at diagnosis, ethnicity, family history, tumor stage, size, grade, or survival, and all the low ER/HER2- tumors with gene expression data available (n=17) were basal-like. Four (9%) of the women with low ER/HER2- tumors had pathogenic mutations in BRCA1 or BRCA2; an additional 4 (9%) had pathogenic mutations in the cancer predisposition genes BLM (n=2), FH (n=1), and MUTYH (n=1). Seven others had VUS in BRCA2, BRIP1, CHEK2, MSH2, MSH6, and SMAD4. Frequency of pathogenic mutations did not differ significantly (P=0.139) between low-staining tumors (18%) and TNBC (13%). Discussion: Low ER/HER2- tumors share similar clinicopathologic characteristics and outcomes with TNBC and have a significant genetic component, with 9% of patients having BRCA1 or BRCA2 germline mutations and an additional 9% having mutations in other cancer-predisposing genes. These data suggest that while ASCO/CAP guidelines define low ER tumors as ER positive, women with low ER/HER2- tumors may benefit from the clinical care and genetic testing provided to women with TNBC. The views expressed in this article are those of the author and do not reflect the official policy of the Department of Army/Navy/Air Force, the Department of Defense, or the U.S. Government. Citation Format: Leann A. Lovejoy, Craig D. Shriver, Rachel E. Ellsworth. Genetic etiology of ER low/HER2- breast tumors [abstract]. In: Proceedings of the AACR Special Conference: Advances in Breast Cancer Research; 2017 Oct 7-10; Hollywood, CA. Philadelphia (PA): AACR; Mol Cancer Res 2018;16(8_Suppl):Abstract nr B51.

Abstract PR08: Longitudinal sampling of ctDNA reveals actionable mutations to optimize treatment of patients with high-grade serous ovarian cancer

Abstract Tumor cells leak their DNA into the blood stream, which allows us to use plasma sample from a patient to measure biomarker levels. In particular, circulating tumor DNA (ctDNA) sampling together with next-generation sequencing technologies provide a rapid and noninvasive test for quantifying tumor response to a treatment as well as finding potentially actionable mutations. The main objective of our study was to use ctDNA to establish a therapeutic window for targeting potentially actionable mutations to optimize the treatment of high-grade serous ovarian cancer (HGSOC) patients. We employed a targeted sequencing panel of 508 clinically annotated cancer genes to screen for actionable copy-number variations (CNV) and single-nucleotide variants (SNV) in 56 plasma and 20 fresh tumor samples from 13 patients with stage IIB-IVB HGSOC treated at the Turku University Hospital, Finland. All patients were surgically debulked and had received standard first-line carboplatin and paclitaxel chemotherapy. We analyzed longitudinal samples (from 3-6 time points per patient) from 11 patients during primary therapy, and from two patients with multiple (two and three samples per patient) relapsed disease. DNA isolated from tumor tissue and plasma was analyzed for genetic alterations by targeted deep-sequencing. White blood cell DNA was used as germline controls. Pathogenic germline mutations in 14 known ovarian cancer susceptibility genes were evaluated with CADD annotation framework and ClinVar database. Minimum requirements for passed somatic SNVs were CADD phred score 10, sequencing depth 100 (ctDNA) or 30 reads (tumor and blood), variant read count 4 or frequency 0.001. Variants detected in blood with &amp;gt; 2 reads were excluded. Additionally, a variant that passed filtering in at least one sample of a patient was included, regardless of threshold, if present in any other specimen. Thresholds of &amp;lt; 0.7 and &amp;gt; 4 copies were used as functionally relevant losses and gains, respectively. Clinically relevant germline pathogenic mutations in BRCA2, ATR, APC, and RB1 genes were identified from blood samples of five (38%) patients. The preoperative samples were also characterized by the highest somatic mutation burden, with a median total variant allele frequency (VAF) of 0.16 (range 0.02-0.77). Importantly, clinically relevant CNVs were detected in preoperative ctDNA of 6 (46%) patients, in potentially targetable driver genes such as ERBB2, PIK3CA, and CDK12. In six patients undergoing neoadjuvant chemotherapy (NACT), we noted a significant decrease in total ctDNA mutation burden during treatment, consistent with a drop in CA-125 levels. Interestingly, the levels of CNV counts increased in ctDNA during NACT in two patients, which associated with improved response to chemotherapy. During adjuvant treatment after debulking surgery, potentially actionable mutations were detected in all patients, with median total VAF of 0.03 (range 0.006-0.43). Importantly, in patients with no macroscopic residual disease, we detected a median of 2.5 (range 1-21) mutations in adjuvant treatment samples, indicating a molecularly persistent disease. Further, a subset of these alterations in, e.g., FANCA and PRCKB persisted during adjuvant treatment, revealing a potential therapeutic window for targeted treatments. Finally, two patients with progressive disease were characterized by predominantly either ctDNA gains or losses. Further, potentially actionable mutations in, e.g., TNXRD1, which sensitizes to AKT inhibitors, could be identified and tracked during disease progression. Taken together, longitudinal analysis of a panel-based ctDNA targeted sequencing can reliably detect very low VAF alterations, which can be used to identify and track actionable genomic alterations. Thus, ctDNA analysis can open a therapeutic window to target molecular residual disease in patients with newly diagnosed and advanced HGSOC. This abstract is also being presented as Poster B06. Citation Format: Kaiyang Zhang, Liina Salminen, Jaana Oikkonen, Kaisa Huhtinen, Johanna Hynninen, Seija Grénman, Sakari Hietanen, Rainer Lehtonen, Anniina Färkkilä, Sampsa Hautaniemi. Longitudinal sampling of ctDNA reveals actionable mutations to optimize treatment of patients with high-grade serous ovarian cancer. [abstract]. In: Proceedings of the AACR Conference: Addressing Critical Questions in Ovarian Cancer Research and Treatment; Oct 1-4, 2017; Pittsburgh, PA. Philadelphia (PA): AACR; Clin Cancer Res 2018;24(15_Suppl):Abstract nr PR08.

DNA Repair Gene Alterations and PARP Inhibitor Response in Patients With Metastatic Castration-Resistant Prostate Cancer.

Background: PARP inhibition is a promising therapeutic strategy for the treatment of men with metastatic castration-resistant prostate cancer whose tumors harbor homologous recombination DNA repair gene alterations. However, questions remain for many practicing clinicians about which patients are ideally suited for PARP inhibitor treatment. This report details our institutional experience using PARP inhibitor therapy in patients whose tumors harbored specific DNA repair gene alterations. Patients and Methods: We performed a retrospective chart review to identify patients at Oregon Health & Science University who were treated with PARP inhibition. We identified 8 patients and determined the impact of the specific DNA repair gene alterations on tumor response and time on treatment with PARP inhibition. Results: A number of DNA repair gene alterations were identified. Three patients had pathogenic BRCA2 mutations and one had a BRCA2 mutation of uncertain significance. Conversely, the 4 other patients' tumors harbored alterations in other DNA repair genes, none of which were clearly pathogenic. A statistically significant difference in benefit was seen between patients whose tumors harbored BRCA2 gene alterations and those whose tumors did not, as measured by >50% decline in prostate-specific antigen levels (100% vs 0%; P=.03) and duration on therapy (31.4 vs 6.4 weeks; P=.03). Conclusions: Our results demonstrate that not all DNA repair alterations are equally predictive of PARP inhibitor response. Importantly, all responding patients had tumors harboring BRCA2 DNA repair alterations, including one without a known pathogenic mutation. Conversely, among the 4 nonresponders, several DNA repair alterations in genes other than BRCA2 were identified that were not clearly pathogenic. This demonstrates the need to carefully examine the functional relevance of the DNA repair alterations identified, especially in genes other than BRCA2, when considering patients for PARP inhibitor treatment.

Abstract IA19: Homologous recombination repair deficiency and breast cancer progression in high-risk populations

Abstract Analysis of cancer genomes has rapidly become an integral part of the practice of clinical oncology, with implications for diagnosis, prognosis, treatment, and prevention. Inherited and sporadic cancers often share common mutational events. When inherited mutations are identified, genetic counseling is an essential component of care. Pathogenic BRCA1 and BRCA2 mutations are the strongest predictors of breast cancer risk and may be the strongest predictors of other inherited forms of solid tumors and hematologic malignancies as well. Waiting to treat advanced breast cancer with targeted therapies is a failure of primary prevention, and population-based strategies for risk management in high-risk populations will be needed. Understanding breast cancer progression in genetic defined subgroups has the potential to accelerate progress in precision medicine. I will discuss ongoing research in our group, our recent findings in defining the genomic landscape of early-onset breast cancer, and future directions for the early detection, treatment, and prevention of breast cancer in high-risk populations. Citation Format: Olufunmilayo I. Olopade. Homologous recombination repair deficiency and breast cancer progression in high-risk populations [abstract]. In: Proceedings of the AACR Special Conference: Advances in Breast Cancer Research; 2017 Oct 7-10; Hollywood, CA. Philadelphia (PA): AACR; Mol Cancer Res 2018;16(8_Suppl):Abstract nr IA19.

Risk-adapted approach to prostate cancer screening

Mass prostatic specific antigen (PSA) testing (population-based PSA screening) has remained controversial, nevertheless there are men cohorts likely to benefit from PSA screening. Heritable factors contribute to 60 % risk for developing familial prostate cancer. Despite the fact that its clinical application is challenging due to polygenic inheritance, advances in new generation sequencing technologies permit identifying highly penetrant germline mutations in genes BRCA1, BRCA2, CHEK2, HOXB13 and MMR associated with tremendous increase in risk of developing the prostate cancer. Several germline mutations are associated with clinically aggressiveness of disease and shortened survival. Targeted screening that is based on family history and genomic aberrations should be the next step towards the precision medicine. Men at elevated risk should been performed for early detection are those with familiar history of prostate cancer, or BRCA1, BRCA2, CHEK2, HOXB13 and MMR pathogenic germline mutation carriers, or first line relatives diagnosed with certain types of cancer. Systematic PSA testing in 1–2 years among germline mutation carriers men beginning at age 45 years would contribute to increase in early detection of localized prostate cancer resulting in more chance of curative treatment and improve survival rates

Abstract 1359: Somatic involution of pathogenic BRCA1 germline mutations

Abstract There is currently a wealth of data on the tumor genomic contexture from BRCA1/2 carriers, particularly breast and ovarian carcinomas. By contrast, little attention has as yet been paid to the genomic status of cancer-related normal tissues from these individuals. Here, we investigated the status of pathogenic BRCA1 germline mutations (GM) in breast (B) and gynecologic (GYN) tissues. Methods: We examined 121 DNA samples (48 B; 36 GYN; 37 tumors) from an equal number of paraffin blocks obtained upon prophylactic or debulking surgery from 44 BRCA1/2 carriers (mean age 38 yrs, range 24-62; 43 BRCA1 carriers). Six women had never had cancer manifestation (CM). At the time of surgery, 32 were cancer-free but had received neo- or adjuvant chemotherapy, and 6 had concurrent cancer without prior treatment. Following multimethod DNA quality control, mutation validation and sample identity match to exclude false negatives, we interrogated GM presence in tissues in comparison to clinicopathologic data and tumor genotypes (60-gene panel; mean read depth over 800). Results: In 19 samples from 13 BRCA1 carriers, including 13 normal B/GYN and 6 tumors, the germline mutation was present at frequencies lower than 5% (observed with the integrated genome viewer) up to 12%, or it was undetectable with Sanger sequencing and multiplex PCR. This condition, termed GM-loss, was present in 13 GYN, 12 of which in the histologically normal tube, and in only 3 B with fibrocystic disease (p=0.0210). It was also present in one ovarian thecoma but it was absent in usual or atypical hyperplasia in B. GM-loss was observed in 9/55 normal tissues from breast cancer and in 4/6 normal tissues from ovarian cancer patients, while it was absent in the normal tissues from women without CM (p=0.0002). GM-loss mostly affected the BRCA1 BRCT functional domain (p&amp;lt;0.0001) and concerned large deletions (10/17 samples), small indels (5/32; 15.6%) and less so single-nucleotide substitutions (4/66; 6.0%; p&amp;lt;0.0001). Normal tissue GM-loss was rather localized, e.g., in 10 patients with multiple samples it was present in only one B or fallopian tube. It was, however, related to the status of concurrent ovarian tumors in 3/6 cases. In the normal tube of these patients, next to the low-frequency GM a somatic pathogenic BRCA1 mutation was present at 25-41% allelic frequency; the same was observed in one breast tumor after neoadjuvant chemotherapy. Three of the replaced GMs were indels, two corresponded to the well-known BRCA1 p.Q1777fs, and all novel mutations were missense. Conclusions: We observed BRCA1 GM-loss in normal tissues, breast and mostly fallopian tube, in about 30% of the carriers in the present cohort, all with previous or concurrent cancer. The phenomenon seems analogous to the GM reversion described in tumors but, as shown, it may also occur in the absence of prior treatment. Its origin and impact on cancer dynamics and therapeutic approaches seem worth further pursuing with functional studies. Citation Format: Vassiliki Kotoula, Efterpi Demiri, Florentia Fostira, Eleni Vrettou, Kyriaki Papadopoulou, Ioannis Tikas, Konstantinos Papazisis, Thomas Zaramboukas, Asimina Asimaki-Vlachopoulou, Spyridon Miliaras, Elena Fountzilas, Ananias Ananiadis, Sofia Chrisafi, Christos Poulios, Ioannis Natsiopoulos, Aris Tsiftsoglou, George Fountzilas. Somatic involution of pathogenic BRCA1 germline mutations [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1359.

High frequency of pathogenic non-founder germline mutations in BRCA1 and BRCA2 in families with breast and ovarian cancer in a founder population

BackgroundPathogenic BRCA1 founder mutations (c.4035delA, c.5266dupC) contribute to 3.77% of all consecutive primary breast cancers and 9.9% of all consecutive primary ovarian cancers. Identifying germline pathogenic gene variants in patients with primary breast and ovarian cancer could significantly impact the medical management of patients. The aim of the study was to evaluate the rate of pathogenic mutations in the 26 breast and ovarian cancer susceptibility genes in patients who meet the criteria for BRCA1/2 testing and to compare the accuracy of different selection criteria for second-line testing in a founder population.MethodsFifteen female probands and 1 male proband that met National Comprehensive Cancer Network (NCCN) criteria for BRCA1/2 testing were included in the study and underwent 26-gene panel testing. Fourteen probands had breast cancer, one proband had ovarian cancer, and one proband had both breast and ovarian cancer. In a 26-gene panel, the following breast and/or ovarian cancer susceptibility genes were included: ATM, BARD1, BLM, BRCA1, BRCA2, BRIP1, CDH1, CHEK2, EPCAM, FAM175A, MEN1, MLH1, MRE11A, MSH2, MSH6, MUTYH, NBN, PALB2, PMS2, PTEN, RAD50, RAD51C, RAD51D, STK11, TP53, and XRCC2. All patients previously tested negative for BRCA1 founder mutations.ResultsIn 44% (7 out of 16) of tested probands, pathogenic mutations were identified. Six probands carried pathogenic mutations in BRCA1, and one proband carried pathogenic mutations in BRCA2. In patients, a variant of uncertain significance was found in BRCA2, RAD50, MRE11A and CDH1. The Manchester scoring system showed a high accuracy (87.5%), high sensitivity (85.7%) and high specificity (88.9%) for the prediction of pathogenic non-founder BRCA1/2 mutations.ConclusionA relatively high incidence of pathogenic non-founder BRCA1/2 mutations was observed in a founder population. The Manchester scoring system predicted the probability of non-founder pathogenic mutations with high accuracy.

PO-023 Chloroquine sensitises oncogenic NOTCH1 driven human T cell leukaemia to γ-secretase inhibition

IntroductionFamilial breast cancer (BC) cases account for 5%–10% of all BC cases, and are essentially correlated with the prevalence of pathogenic BRCA1 and BRCA2 mutations in cancer patients. Recent advances in next generation sequencing (NGS) have enabled panel gene testing, or simultaneous testing for mutations in multiple genes for a clinical condition. With more widespread genetic testing, an increased detection of alleles with moderate penetrance without established clinical guidelines and of variants of unknown significance (VUS) as either benign or disease-causing will occur. Functional analyses on VUS may identify pathogenicity, and clearly categorise their mutational status. We carried-out a proof-of concept in vitro functional analysis in peripheral blood lymphocytes of VUS-harbouring individuals and healthy controls assessing the cellular response to γ-radiation.Material and methodsFour samples were collected, one BC patient with a pathogenic ATM mutation; one BRCA2 VUS detected without diagnosed disease, and two controls (A, B, no variant detected). All samples were previously sequenced by NGS to confirm the presence of pathogenic mutations or VUS detected in a genetic testing panel. Several methodologies were selected to evaluate the cellular response to genetic lesions induced by γ-radiation (2Gy): chromosomal aberrations (CA), micronuclei (MN) and comet assay.Results and discussionsComet assay results revealed a higher DNA damage in the patient with a pathogenic ATM mutation (14% DNA in tail) after irradiation, which might be related with the non-optimal function of the ATM protein. The BRCA2 VUS sample also showed a higher%DNA in tail (6.5%), contrasting with the damage quantified for control samples (2.8% and 0.9% DNA in tail for control A and B, respectively). These results revealed that the presence of such variants might be correlated with their biological function, being crucial to categorise the mutational status. Regarding CA and MN, these assays were not sensitive enough to discriminate pathogenic from control samples.ConclusionResults obtained for comet so far suggest a significant difference between variant carriers and controls, which may indicate an increased susceptibility to ionising radiation. However, more sensitive assays (e.g. H2AX) will be necessary allowing more informative results. As this study is a proof-of concept approach, in the near future the sample size will be incremented.

An opportunity lost: Low rates of fertility counseling in pathogenic BRCA mutation carriers

An opportunity lost: Low rates of fertility counseling in pathogenic BRCA mutation carriers

Therapy susceptible germline-related BRCA 1-mutation in a case of metastasized mixed adeno-neuroendocrine carcinoma (MANEC) of the small bowel

BackgroundAdenocarcinomas or combined adeno-neuroendocrine carcinomas (MANEC) of small bowel usually have a dismal prognosis with limited systemic therapy options. This is the first description of a patient showing a germline-related BRCA1 mutated MANEC of his ileum. The tumor presented a susceptibility to a combined chemotherapy and the PARP1-inhibitor olaparib.Case presentationA 74-year old male patient presented with a metastasized MANEC of his ileum. Due to clinical symptoms his ileum-tumor and the single brain metastasis were removed. We verified the same pathogenic (class 5) BRCA1 mutation in different tumor locations. There was no known personal history of a previous malignant tumor. Nevertheless we identified his BRCA1 mutation as germline-related. A systemic treatment was started including Gemcitabine followed by selective internal radiotherapy (SIRT) to treat liver metastases and in the further course Capecitabine but this treatment finally failed after 9 months and all liver metastases showed progression. The treatment failure was the reason to induce an individualized therapeutic approach using combined chemotherapy of carboplatin, paclitaxel and the Poly (ADP-ribose) polymerase- (PARP)-inhibitor olaparib analogous to the treatment protocol of Oza et al.All liver metastases demonstrated with significant tumor regression after 3 months and could be removed. In his most current follow up from December 2017 (25 months after his primary diagnosis) the patient is in a very good general condition without evidence for further metastases.ConclusionWe present first evidence of a therapy susceptible germline-related BRCA1 mutation in small bowel adeno-neuroendocrine carcinoma (MANEC). Our findings offer a personalized treatment option. The germline background was unexpected in a 74-year old man with no previously known tumor burden. We should be aware of the familiar background in tumors of older patients as well.

Genome-wide and high-density CRISPR-Cas9 screens identify point mutations in PARP1 causing PARP inhibitor resistance

Although PARP inhibitors (PARPi) target homologous recombination defective tumours, drug resistance frequently emerges, often via poorly understood mechanisms. Here, using genome-wide and high-density CRISPR-Cas9 “tag-mutate-enrich” mutagenesis screens, we identify close to full-length mutant forms of PARP1 that cause in vitro and in vivo PARPi resistance. Mutations both within and outside of the PARP1 DNA-binding zinc-finger domains cause PARPi resistance and alter PARP1 trapping, as does a PARP1 mutation found in a clinical case of PARPi resistance. This reinforces the importance of trapped PARP1 as a cytotoxic DNA lesion and suggests that PARP1 intramolecular interactions might influence PARPi-mediated cytotoxicity. PARP1 mutations are also tolerated in cells with a pathogenic BRCA1 mutation where they result in distinct sensitivities to chemotherapeutic drugs compared to other mechanisms of PARPi resistance (BRCA1 reversion, 53BP1, REV7 (MAD2L2) mutation), suggesting that the underlying mechanism of PARPi resistance that emerges could influence the success of subsequent therapies.

Complete response to orally administered melphalan in malignant pleural effusion from an occult female genital organ primary neoplasm\xa0with BRCA1/2 mutations: a case report

BackgroundDefinite diagnosis of metastasis from unknown primary depends on a comprehensive immunohistochemical investigation of tumor specimen. Accurate identification of the origin site usually helps a lot in choosing the most appropriate treatment. Molecular characterization provides more chance of a cure. Echoing modern medical development, BRCA1/2 mutations have been correlated with high efficiency of poly(adenosine diphosphate-ribose) polymerase inhibitors in ovarian cancer. While a previous case report demonstrated a surprising cure of platinum-resistant ovarian cancer with BRCA2 mutation by orally administered melphalan.Case presentationA 53-year-old Taiwanese woman’s malignant pleural effusion was diagnosed to be a metastasis from an occult cancer in female genital organ by diligent pathological study despite absence of image evidence. She resolutely refused intravenously administered chemotherapy. After failure of anti-estrogen tamoxifen, orally administered melphalan achieved excellent complete remission. Pathogenic homozygous BRCA1 and BRCA2 mutations were later detected in tumor cells by next-generation sequencing. The same BRCA2 mutation exists in a heterozygous status in the germline deoxyribonucleic acid.ConclusionsThis is so far the second report of long-term remission of advanced female genital organ cancer with BRCA mutations achieved by orally administered melphalan. BRCA1/2 mutations and even all “BRCAness” of malignancy, at least ovarian cancer and ovarian-related cancers, probably not only correlate with high efficacy of poly(adenosine diphosphate-ribose) polymerase inhibitors but also lead to a high-potential cure by orally administered melphalan. We recommend that clinical trials that test this assumption be carefully designed and sophisticatedly performed.

Somatic BRCA1 mutations in clinically sporadic breast cancer with medullary histological features

BackgroundThe role of somatic BRCA1/2 gene mutations in breast cancer is getting increasing attention in view of hereditary disease. The medullary phenotype and triple negative intrinsic subtypes are often, but not exclusively encountered in BRCA1 germline mutated breast cancer, whilst for BRCA2, no association to specific histological features are known. In this study, we addressed the relationship between morphological medullary phenotype and BRCA1/2 somatic mutations in breast cancer without known positive family anamnesis.Methods32 clinically sporadic breast cancers with medullary features were analyzed for somatic BRCA1/2 mutations (all coding exons) with next-generation sequencing technology. Paraffin-embedded formalin-fixed breast cancer samples from all patients were analyzed.ResultsThree of 32 tumors (9%) had pathogenic (ARUP class-5) BRCA1 gene alterations. Two of these pathogenic variants exhibited deletions leading to frameshift mutations (p.Glu23fs, p.Val1234fs), and the remaining single-nucleotid-variant resulted in premature STOP codon (p.Glu60Ter). In one patient, the same pathogenic BRCA1 mutation was detected (p.Glu23fs) in normal breast tissue. Retrospective follow-up in two patients revealed a positive family history for breast cancer and consecutive germline mutation testing confirmed presence of BRCA1 mutations. No somatic pathogenic BRCA2 mutations were detected.ConclusionsBRCA1 mutation testing may be useful in clinically sporadic breast cancer patients with medullary features to identify potential mutation carriers independently from intrinsic molecular subtype. Formalin-fixed paraffin-embedded cancer tissue can undergo testing within a routine molecular-diagnostic setting as a clinical BRCA1/2 mutation screening strategy.

Abstract P4-02-10: Breast cancer surveillance in high-risk women with dynamic contrast-enhanced magnetic resonance imaging every 6 months: Results from a single institution study

Abstract Purpose: To develop a novel approach for early detection of breast cancer and examine molecular features of screen detected cancers in prospectively ascertained high-risk women undergoing semi-annual dynamic contrast-enhanced breast magnetic resonance imaging (DCE-MRI) for women at high genetic risk. Background: Women with a personal or family history of breast cancer and genetic mutation carriers of BRCA1 and BRCA2 have a higher than normal risk of breast cancer. An intensified screening surveillance regimen is an early detection strategy in high-risk women. The American Cancer Society recommends annual DCE-MRI in addition to annual mammogram based off several pivotal screening studies that demonstrated improved sensitivity and cancer detection rates and decreased interval cancer rates with the addition of annual DCE-MRI. Questions remain regarding the optimal screening modality and interval regimen in these high-risk women. Methods: Between 2004 and 2016, we assembled a prospective cohort of high-risk women undergoing semi-annual DCE-MRI and annual mammography. To be eligible, women had a lifetime breast cancer risk &amp;gt;20% and/or tested positive for a pathogenic mutation using a cancer gene panel including BRCA1, BRCA2, CDH1, PALB2, CHEK2 and other cancer susceptibility genes in the DNA repair pathway. Somatic mutation events in screen-detected tumors were investigated using UW-OncoPlex cancer gene panel using DNA extracted from FFPE shavings. Results: 295 women were recruited to the study; 44% of the study participants had pathogenic mutations in BRCA1 or BRCA2 genes. At a median follow-up of 3.3 years (range 0-12 years), 3 DCIS and 13 early stage invasive breast cancers were detected, of which 14 occurred in subjects with identifiable pathogenic mutations (11 BRCA1, 2 BRCA2, 1 CDH1). The incidence rate is 1.3% in all subjects, but 3.5 % per year in BRCA1 carriers. DCE-MRI identified all 13 invasive cancers at a mean size of 0.61 cm (range 0.1-1.0 cm); none had lymph node metastasis. No interval cancers occurred. In addition, 7 of the breast cancers were detected on DCE-MRI imaging obtained at the 6 months screening interval; they would be interval cancers if only annual screening were implemented. There was very little DNA for somatic mutation testing in the majority of cases. However, as expected, there was heterogeneity in the spectrum of mutations but the most commonly somatically mutated gene in the early cancers was TP53. Conclusions: DCE-MRI every 6 months performed well for early detection of invasive breast cancer in high-risk women, accomplishing the ultimate goal of breast cancer screening—detecting node-negative, invasive tumors less than 1 cm. Semi-annual DCE-MRI performed especially well in BRCA1 mutation carriers at risk for the most aggressive subtype of breast cancer. Further interventional studies evaluating this novel screening approach are warranted to personalize breast cancer risk assessment and prevention. Citation Format: Whitaker K, Guindalini R, Abe H, Sheeth D, Huo D, Hong S, Churpek J, Verp M, Obeid E, Zheng Y, Amico A, Yoshimatsu T, Olopade O. Breast cancer surveillance in high-risk women with dynamic contrast-enhanced magnetic resonance imaging every 6 months: Results from a single institution study [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P4-02-10.

Abstract P5-19-04: Psychosocial impact of a multi-modality surveillance program for women at high-risk for breast cancer

Abstract Purpose: To evaluate the psychosocial impact of semi-annual dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) screening in women at high-risk for breast cancer. Background: For women with BRCA1 and BRCA2 mutations and/or a personal or family history of breast cancer, annual breast MRI has shown improved sensitivity and cancer detection compared to mammography. However, MRI's heightened sensitivity may lead to increased: false positives requiring additional follow-up biopsy/imaging; iatrogenic risk; and psychosocial distress, which all may negatively impact women's overall health-related quality of life. Methods: Between 2004 and 2016, we assembled a prospective cohort of high-risk women undergoing semi-annual DCE-MRI and annual mammography. We reviewed a subset of this group. Participants completed psychosocial assessments at baseline and 6-month visits using the following measures: coping (MBSS); state/trait anxiety (STAI-S/T); depression (BDI-II); risk perception; and mental health (SF-36). Participants were classified according to Monitor or Blunter coping style. Mixed-effects logistic regressions models examined effects of demographics on psychosocial changes over time. Results: 295 women were recruited to the study; 44% of the study participants had pathogenic mutations in BRCA1 or BRCA2 genes. 232 of 295 enrolled participants (78.6%) completed psychosocial assessments. For the total population: median age 44y (range: 21-73), 71% ≥college/post-graduate education; 84% Caucasian; 8% African American; 2% Latino; 99% with health insurance; 72% annual income of &amp;gt;$60,000. One third of women had a personal cancer history. Participants were evenly split between baseline Monitoring and Blunting coping style (49% and 51%, respectively). No significant differences were found between demographics (age, race, income, mutation, cancer type, cancer history) or psychosocial factors (baseline trait anxiety (p =0.64), depression (p =0.65), SF36 global health (p=0.66). After adjusting for education, race, cancer history and coping, women with ≥$60,000 income had lower trait anxiety (p&amp;lt;0.000) and greater mental health (p&amp;lt;0.001) than those with &amp;lt;$60,000 income. Over time, change in trait anxiety varied by coping (p=0.0006): Blunters did not experience significant changes in trait anxiety (p=0.072) while Monitors had significant diminished trait anxiety over time (p&amp;lt;0.001). For depression, women with ≥$60,000 income and college educated had lower BDI-II depression (p&amp;lt;0.000). Yet, women with a cancer history had significantly greater BDH-II depression (p= 0.048). Mental health over time varied by race as non-whites had greater gains in mental health (p=0.001) over time than whites (p=0.03). Conclusion: Semi-annual DCE-MRI did not cause a significantly elevated state anxiety or depression, nor was there a significant decline in mental health over time for groups regardless of cancer history and genetic mutation status. Coping style may have an impact on psychosocial outcomes for those undergoing heightened surveillance over time. Citation Format: Amico AL, Fang R, Raoul A, Wroblewski K, Nielsen S, Weipert C, Abe H, Sheth D, Romero I, Kulkarni K, Schacht D, Patrick-Miller L, Verp M, Bradbury AR, Hlubocky F, Olopade OI. Psychosocial impact of a multi-modality surveillance program for women at high-risk for breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P5-19-04.

Abstract OT3-01-01: Chemoprevention in BRCA1 mutation carriers (CIBRAC): A proof of concept clinical trial

Abstract Introduction Currently, risk management options for BRCA1 mutation carriers include risk-reducing mastectomies (RRM), and selective oestrogen receptor modulators (SERMs). However, the majority of BRCA1-associated tumours are oestrogen receptor negative (ER-ve). SERMs do not reduce the incidence of ER-ve tumours, therefore are unlikely to be effective in this group of women. Oestrogen and its metabolites have been shown to cause DNA damage, resulting in genomic instability in BRCA1 deficient breast cells, an early hallmark of BRCA1-related cancers. Based on this, we hypothesise that oestrogen suppression in BRCA1 carriers may reduce DNA damage in breast tissue, and thereby potentially reducing breast cancer risk. Trial Design The study is an interventional, non-randomised, crossover study without masking. The primary purpose is to assess the impact of these drugs on suppression of DNA damage and therefore their potential for use as chemopreventive agents in this setting. Pre-menopausal BRCA1 mutation carriers will be recruited from the Family History Clinic at Belfast City Hospital. The proportion of women who receive information packs who progress to trial entry will be recorded, with target recruitment of twelve women to the trial. Participants will complete quality of life questionnaires, provide blood and urine samples and undergo baseline ultrasound-guided breast biopsy. Half of these women will receive a SERM (tamoxifen) for 3 months, whilst the other half receives oestrogen suppression therapy (goserelin and anastrazole), and then provide repeat questionnaires and samples (blood, urine and breast biopsy). Treatment groups will cross over for a further 3 months treatment, with a final set of questionnaires and samples taken. Compliance with treatment and adverse events will be monitored throughout the study. Eligibility Criteria Inclusion criteria: · Female · Age ≥18 years · Premenopausal · Known pathogenic BRCA1 mutation · Intact ovaries · No previous breast/ovarian/other carcinoma · No previous use of chemoprevention · Willingness to use non-hormonal methods of contraception Exclusion criteria: · BRCA1 mutation of uncertain significance · Contraindications to study drugs or breast biopsies · Pregnancy or breastfeeding · Inability to give informed consent · Patient awaiting risk reducing surgery Specific Aims The primary objective is to assess the feasibility of treatments by measuring successful recruitment rates and compliance. The secondary objective is to establish tolerability of interventions through quality of life measurements and adverse event occurrence. Exploratory objectives will assess the potential of treatments to reduce DNA damage through analysis of breast biopsies using comet assays and immunohistochemistry for 53BP1 and γH2AX (markers of DNA double-strand breaks). Oestrogen and metabolite levels will be measured in blood and urine samples using UPLC-MS/MS. Statistical Methods Not applicable due to nature and size of pilot study Present accrual: 1 participant Target accrual: 12 participants Contact information for people with specific interest in trial: aideen.campbell@qub.ac.uk or s.mcintosh@qub.ac.uk Citation Format: Campbell AM, Savage KI, McIntosh SA. Chemoprevention in BRCA1 mutation carriers (CIBRAC): A proof of concept clinical trial [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr OT3-01-01.

Abstract P4-07-01: Assessment of the hereditary component in 94 cancer predisposition genes to triple negative breast cancer

Abstract Background: In women with triple negative breast cancer (TNBC) unselected for age or family history, 8-14% and 5% of patients harbor germline mutations in BRCA1 and BRCA2, respectively. Diagnosis of TNBC &amp;lt;60 years of age is one of the NCCN criterion for genetic testing of BRCA1 and BRCA2. The contribution of germline mutations in other cancer predisposition genes to TNBC, is, however, not well-studied. Methods: TNBC was classified as tumors with &amp;lt;1% positively staining cells for ER and PR and HER2 = 0+, 1+ or 2+/not amplified. Genomic DNA was isolated from blood samples and targeted sequencing was performed using the TruSight Cancer panel (Illumina). Pathogenic mutations were identified using VariantStudio and classified as pathogenic, uncertain significance (VUS) or benign using ClinVar. Results: 196 female patients diagnosed with TNBC 2001-2014 had genomic DNA available. Average age at diagnosis was 52.8 years (range 34.1-83.4 years). The majority of patients were of European (66%) or African (31%) American ancestry; 26% had a family history and 13% had died of disease with an average time to death of 2.81 years. Twenty-three (12%) of women with TNBC had pathogenic mutations in breast cancer genes BRCA1 (n=14), BRCA2 (n=5), PALB2 (n=1) and CHEK2 (n=3), two women had mutations in the colon cancer genes MUTYH, one had a mutation in the ovarian cancer gene BRIP1, and an additional three women had pathogenic mutations in cancer predisposition genes FANCD2, SDHB and XPC. An additional 42 women had VUS in 20 genes, including one in BRCA1 and 5 in BRCA2. Discussion: Although the majority of pathogenic mutations in this cohort of women with TNBC were in the BRCA1 and BRCA2 genes (10%), panel testing allowed for the detection of mutations in other breast (2%), colon (1%), ovarian (1%) and other cancer (2%) predisposition genes. Panel testing thus identifies genes other than BRCA1/2 associated with increased risk of TNBC and may incidentally identify women who would benefit from enhanced surveillance for other cancers. The opinions or assertions contained herein are the private ones of the author/speaker and are not to be construed as official or reflecting the views of the Department of Defense, the Uniformed Services University of the Health Sciences or any other agency of the U.S. Government. Citation Format: Ellsworth RE, Lovejoy LA, Shriver CD. Assessment of the hereditary component in 94 cancer predisposition genes to triple negative breast cancer [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P4-07-01.