Mutations In Ovarian Cancer Patients Research Articles

Next generation sequencing analysis of BRCA1/2 genes reveals clinically significant pathogenic mutations in ovarian cancer patients

Next generation sequencing analysis of BRCA1/2 genes reveals clinically significant pathogenic mutations in ovarian cancer patients

Mutational Profile and Dynamics of PPM1D-Mutant Clones in the Spectrum of Myeloid Disorders

Introduction Mutations in PPM1D a regulator of DNA damage response, are enriched in patients with clonal hematopoiesis (CH) exposed to cytotoxic treatment or with therapy-related myeloid neoplasm. However, their role in leukemic transformation is unclear. Here, we describe a large cohort of patients with PPM1D mutations from CH to acute myeloid leukemia (AML) to understand the molecular landscape of mutant PPM1D related disease and the longitudinal dynamics of CH under treatment. Methods We included, in a non-sequential cohort, 96 PPM1D mutated patients treated at Gustave Roussy with CH, clonal cytopenia of unknown significance (CCUS), myelodysplastic syndrome (MDS), myeloproliferative neoplasm (MPN) or AML. A 77 gene panel NGS analysis was performed. An additional 16 PPM1D mutated AML patients from ALFA trials (1200, 0701,0702) were included. 10 patients with ovarian cancer from this cohort had sequential blood samples (OvBIOMark trial) available prior to hematologic evaluation, from the diagnosis or first relapse of their cancer through the course of therapy. Those samples were analyzed with a UMI based 18 gene panel (HaloplexHS, Agilent). Overall survival (OS) analyses were performed with the Kaplan-Meier method from the time of diagnosis until death from any cause. Results Among the 112 patients with PPM1D mutations, 23 (21%) had CH, 36 (32%) CCUS, 19 (17%) MDS, 25 (22%) AML (including 4 relapses), and 9 (8%) MPN. Median age was 65 [range, 21-88] years with 67% of females. Only 18% of the patients had no previous cancer history. Most frequent primary cancers were gynecological cancer (27%), lymphoid malignancies (22%), and breast cancer (17%). Median time between primary cancer diagnosis and hematologic assessment was 5.3 [range, 1.2-8.5] years. Eighty six percent of patients had one PPM1D mutation, 9% had 2, and 5% had 3 or more (restricted to CH and CCUS). Median variant allele frequency (VAF) was 3% [0.2-38], 3% [1-31], 2.4% [1-41], 23% [1-50], and 4% [1-42] in CH, CCUS, MDS, AML and MPN, respectively. Among CH/CCUS patients, PPM1D was the sole detected somatic mutation in 39% (23/59), compared to 9% (5/53) in MDS/AML/MPN patients (odds ratio=6, p=0.0004); 3/5 of the latter had complex karyotype. The most frequently co-mutated genes were DNMT3A (29%) and TP53 (25%), uniformly across all conditions ( Fig 1A). MDS-related gene mutations, RUNX1 (7%), ASXL1 (4%), SF3B1 (4%), SRSF2 (4%), U2AF1 (4%), were specific to AML/MDS/MPN. Among 28 patients with both PPM1D and TP53 mutations, PPM1D mutations were dominant or co-dominant in 64% (18/28), and secondary in 36% (10/28). IPSS-M risk of evaluable MDS was high/very high in 54% (7/13) of patients. ELN 2022 classification of AML was adverse in 68% (17/25) of patients. AML treatment options included best supportive care for 8% of patients (2), 5-Azacytidine for 32% (8), intensive chemotherapy for 60% (15). With a median follow up of 2.4 years, the median OS was 4.6 (CI95%; 4.1-NA), 1.31 (CI95%; 0.53-NA), 1.27 (0.43-NA), 0.66 (CI95%; 0.42-1.26) and 20.4 (CI95%; 20.4-NA) years for CH, CCUS, MDS, AML and MPN, respectively. TP53 mutation status did not stratify OS. Four ovarian cancer patients with CH/CCUS transformed to MDS/AML with a median of 5.3 [1-12] years. At transformation, 3/4 had a stable PPM1D mutation, 1/4 an increase in PPM1D VAF (2 to 28%), and 3/4 had TP53 mutations before and at transformation. We analyzed 67 timepoints from 10 ovarian cancer patients during therapy (median 7 per patient). 10/10 received alkylating agents and 5/10 PARPi. The median number of mutations per patient was 4 and 6 at baseline and last follow-up, respectively. PPM1D-mutated clone size increased from 0.4% [0.1-3] at baseline to 7% [2-30] at last follow-up. Beyond clonal expansion, PPM1D VAF dynamics showed non-linear changes related to alkylating agents exposure: 2/10 patients had a continuous expansion, 2/10 had expansion then contraction (GR-1/3, Fig 1B), and 6/10 had expansion then stabilization (GR-2, Fig 1B). Conclusion We described a large cohort of PPM1D mutated patients from CH to AML. Their prognosis was poor independently of TP53 mutation at AML/MDS stage. PPM1D mutations were frequently part of the dominant clone. To confirm the clonal architecture, single cell sequencing analysis in 8 AML/MDS patients are ongoing. Trajectory of PPM1D mutations in ovarian cancer patients revealed a non-linear alkylating agent dependency which warrants further investigation.

Frequency of BRCA1 and BRCA2 mutations in ovarian cancer patients in South-East Poland

BackgroundMutations in BRCA1 and BRCA2 genes are well-established risk factors of breast and ovarian cancer. In our former study, we observed that approximately 6% of unselected ovarian cancer patients in the region of Podkarpacie (South-East Poland) carry BRCA1 causative founder variants, which is significantly lower than in other regions of Poland. Therefore, it is deeply justified to do research based on the sequencing of whole BRCA1 and BRCA2 genes.MethodsWe examined 158 consecutive unselected cases of ovarian cancer patients from the region of Podkarpacie. We performed BRCA1 and BRCA2 genes Next-Generation Sequencing study in all cases.ResultsAltogether, in 18 of 158 (11.4%) ovarian cancer patients with BRCA1 or BRCA2 pathogenic mutations were found. BRCA1 pathogenic variants were detected in 11 of the 158 (7.0%) ovarian cancer cases. 10 of 11 (91%) detected BRCA1 mutations were founder mutations, detectable with the standard test used in Poland. BRCA2 pathogenic variants were found in 7 of the 158 (4.4%) cases. No BRCA2 pathogenic variants were founder mutations. The median age of patients at the diagnosis of the 18 hereditary ovarian cancers was 57.5 years.ConclusionsThe frequency of BRCA1 or BRCA2 gene mutation carriers among patients with ovarian cancer from the Podkarpacie region is comparable to other regions of Poland. However, a significantly higher percentage of BRCA2 gene mutations was observed, that were not detectable with a standard test for detection of founder mutations. Diagnostics based only on testing the BRCA1/2 Polish founder mutations is characterized by relatively low sensitivity in the case of ovarian cancer patients from South-East Poland and should be supplemented by NGS study, in particular of the BRCA2 gene.

BRCA1 and BRCA2 mutations in ovarian cancer patients from Belarus: update

BackgroundMutations in BRCA1 and BRCA2 are well-established risk factors for breast and ovarian cancer. In Central-Eastern European counties, the founder mutations in the BRCA1 are responsible for a significant proportion of ovarian cancer cases, however, regional differences in the frequencies of various mutations may exist. The spectrum and frequency of BRCA1/2 mutations between ovarian cancer patients have not yet been precisely established in Belarus.MethodsTwo hundred fourteen consecutive unselected cases of ovarian cancer patients from the region of West Belarus were examined. We studied 13 founder mutations in BRCA1 (c.5266dupC, c.4035delA, c.5251C > T, c.181 T > G, c.676delT, c.68_69delAG, c.3700_3704delGTAAA, c.1687C > T, c.3756_3759delGTCT) and in BRCA2 (c.658_659delGT, c.7913_7917delTTCCT, c.3847_3848delGT, c.5946delT) characteristic for Central European population.ResultsA BRCA1 or BRCA2 founder mutations were detected in 54 of the 214 (25.2%) ovarian cancer cases. The BRCA1 c.5266dupC mutation was detected in 28 patients, followed by c.4035delA mutation observed in 18 patients. BRCA1 c.3756_3759delGTCT, c.68_69delAG, and c.1687C > T were found in 3, 2, and 1 women, respectively. BRCA2 c.658_659delGT mutation was detected in 2 ovarian cancer patients. The median age of diagnosis of the 54 hereditary ovarian cancers was 57.5 years.ConclusionsThe frequency of 13 causative BRCA1 and BRCA2 founder mutations in West Belarus was higher than in other Slavic countries. Testing of BRCA1 (c.5266dupC, c.4035delA, c.3756_3759delGTCT, c.68_69delAG, c.1687C > T as well as c.181 T > G) and BRCA2 (c.658_659delGT) mutations should be considered an inexpensive and sensitive test panel for this population.

Magnitude of benefit of the addition of poly ADP-ribose polymerase (PARP) inhibitors to therapy for malignant tumor: A meta-analysis.

The purpose of this study was to analyze the efficacy of PARP inhibitor on solid tumors. For this study, the following databases were searched for articles published from its inception until July 2019: PubMed, Web of Science, EBSCO, and Cochrane library, of which the main conclusion was the overall survival (OS) and progression-free survival (PFS). We conducted a meta-analysis and the results showed that PARP inhibitor increased the patients' PFS (HR: 0.51, p < 0.001), PFS with BRCA1/2 mutations (HR: 0.32, p < 0.001), OS (HR: 0.74, p < 0.001), OS with BRCA1/2 mutations (HR: 0.78, p = 0.03), complete response (CR) (RR: 1.89, p = 0.10), partial response (PR) (RR: 1.34, p = 0.01), overall response rate (ORR) (RR: 1.42, p = 0.001) respectively. The main adverse events (AEs) observed were decreased appetite. PARP inhibitors may prolong survival. PARP inhibitors were more favorable for BRCA1/2 mutations in ovarian cancer patients. Additionally, the overall safety factor was controllable.

An optimized BRCA1/2 next-generation sequencing for different clinical sample types.

ObjectiveA simultaneous detection of germline and somatic mutations in ovarian cancer (OC) using tumor materials is considered to be cost-effective for BRCA1/2 testing. However, there are limited studies of the analytical performances according to various sample types. The aim of this study is to propose a strategy for routine BRCA1/2 next-generation sequencing (NGS) screening based on analytical performance according to different sample types.MethodsWe compared BRCA1/2 NGS screening assay using buffy coat, fresh-frozen (FF) and formalin-fixed paraffin-embedded (FFPE) from 130 samples.ResultsThe rate of repeated tests in a total of buffy coat, FF and FFPE was 0%, 8%, and 34%, respectively. The accuracy of BRCA1/2 NGS testing was 100.0%, 99.9% and 99.9% in buffy coat, FFPE and FF, respectively. However, due to the presence of variant allele frequency (VAF) shifted heterozygous variants, tumor materials (FFPE and FF) showed lower sensitivity (95.5%–99.0%) than buffy coat (100%). Furthermore, FFPE showed 51.4% of the positive predictive value (PPV) on account of sequence artifacts. When performed in the post-filtration process, PPV was increased by approximately 20% in FFPE. Buffy coat showed 100% of sensitivity, specificity and accuracy in BRCA1/2 NGS test.ConclusionsOn the comparison of the analytical performance according to different sample types, the buffy coat was not affected by sequencing artifacts and VAF shifted variants. Therefore, the blood test should be given priority in detecting germline BRCA1/2 mutation, and tumor materials could be suitable to detect somatic mutations in OC patients without identifying germline BRCA1/2 mutation.

226O - Prevalence of germline BRCA1 and BRCA2 mutations and variants among ovarian, primary peritoneal and fallopian tube cancer patients: A multicentre Indian study

BackgroundThe worldwide reported prevalence of germline pathogenic BRCA1 and BRCA2 mutations in ovarian cancer patients is 17 % (3-27 %) and that of variants of uncertain significance (VUS) is about 10 %. 1, 2There is deficient data on the prevalence of BRCA mutations in Indian patients. We aimed to estimate the prevalence of germline BRCA mutations among Indian patients with ovarian, primary peritoneal or fallopian tube cancer. MethodsThis was a cross-sectional, multicenter, prospective, observational study conducted at 9 sites from different geographical regions across India which enrolled patients with new or previous diagnosis of ovarian, primary peritoneal, or fallopian tube cancer. Patients underwent blood testing using NGS based platform for germline BRCA. The calculated sample size for this study was 228, assuming 5 % precision around an estimated point prevalence of 15.8 % with a dropout rate of 10 %. ResultsA total of 239 patients were enrolled, of whom 16 were newly diagnosed, treatment naïve, and 223 had a previous diagnosis of ovarian cancer and had received between 1- 9 prior lines of therapy. Germline pathogenic or likely pathogenic BRCA1/BRCA2 mutations were seen in 61 (25.5%, 95% CI 20.12, 31.54) and VUS were seen in 10 (4.2%, 95% CI 2.02 – 7.54) patients, respectively. Among 159 patients with serous histology tumours, 48 (30.2%) had pathogenic/likely pathogenic germline BRCA1/BRCA2 mutations while 16 of 74 (16.2 %) patients with non-serous histology tumours had this finding. Of the 61 patients in this study with germline pathogenic/likely pathogenic BRCA1/BRCA2 mutation, 41 (67.1 %) did not have a family history of breast and/or ovarian cancer. ConclusionsApproximately one-fourth of unselected ovarian cancer patients from India had germline BRCA1/BRCA2 mutations of whom, nearly two-thirds did not have a family history of breast and/or ovarian cancers. Legal entity responsible for the studyAstraZeneca Pharma India Limited. FundingAstraZeneca Pharma India Limited. DisclosureS. Gupta: Honoraria (institution), Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca. S. Rajappa: Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca. S.H. Advani: Advisory / Consultancy: AstraZeneca. A. Agarwal: Speaker Bureau / Expert testimony: AstraZeneca. S. Aggarwal: Advisory / Consultancy: AstraZeneca. C. Goswami: Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca. S. Dattatreya Palanki: Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca. D. Arya: Advisory / Consultancy, Speaker Bureau / Expert testimony: AstraZeneca. R. Kodagali: Full / Part-time employment: AstraZeneca. All other authors have declared no conflicts of interest.

LBA11 - Mainstreaming genetic counselling for genetic testing of BRCA1/2 in ovarian cancer patients in Malaysia (MaGIC study)

BackgroundIdentification of germline BRCA mutations in ovarian cancer patients allows clinicians to make treatment decisions and manage risks. Due to the lack of genetic counsellors, clinical geneticists and awareness among clinicians, there is minimal testing regionally. Mainstreaming genetic counselling may improve this access. This study aims to determine the prevalence of BRCA mutations, feasibility of mainstreaming and the psychosocial impact of genetic testing in Malaysian ovarian cancer patients. MethodsThis study aimed to recruit 800 patients with non-mucinous epithelial ovarian, peritoneal or fallopian tube cancer regardless of age or family history, via mainstreaming or referral to genetic services. Genetic Counselling Satisfaction Scale (GCSS), Decisional Conflict Scale (DCS), Psychosocial Aspect of Hereditary Cancer (PAHC) and Cancer Worry Scale (CWS) were used to measure the feasibility and psychosocial outcomes. Mainstreaming clinicians provided feedback through a survey. Results23/28 mainstreaming sites remained active and 47/68 trained clinicians continued to provide genetic counselling. 800 patients have been recruited, 690 via mainstreaming and 110 via genetics route. 790 have been tested, 110 (13.9%) have pathogenic variants, 90 (11.4%) VUS and 590 (74.7%) negative. Majority of the patients were satisfied with their pre-test GC (mean: 23.9; SD: 2.7) and post result disclosure (mean: 24.6; SD: 2.6). Large majority (86%) did not feel conflicted. Patients with no issues in 5 out of 6 PAHC domains increased from 46% at pre-test GC to 62% at result disclosure. However, only 16% of participants showed high distress level after result disclosure, 22% of these were carriers. Generally, there were no significant differences between mainstreaming and genetics pathways. 36/47 (80%) of clinician are confident to counsel patients and want to integrate BRCA testing into their clinical practice. ConclusionsBRCA prevalence for ovarian cancer in Malaysia is 13.9%. There is satisfaction in counselling services, acceptable psychosocial impact with no significant differences between mainstreaming and genetics pathway. Mainstreaming may improve access to genetic testing in Malaysia. Legal entity responsible for the studyCancer Research Malaysia. FundingAstraZeneca. DisclosureS.Y.Y. Yoon: Research grant / Funding (institution), Investigator Initiated Research Grant: AstraZeneca. A. George: Research grant / Funding (institution): Wellcome Trust; Honoraria (institution): AstraZeneca; Honoraria (institution): Roche; Honoraria (institution): Tesaro.

Frequency of germline mutations in BRCA1 and BRCA2 in ovarian cancer patients and their effect on treatment outcome.

Aim of workReporting the incidence and the variants of BRCA1/2 mutations in ovarian cancer patients exploring their effects on the treatment outcomes.Patients and methodsIn total, 104 patients with epithelial ovarian cancer were prospectively recruited to the study. Analysis consisted of the sequencing of all the translated exons and immediately adjacent intronic regions of the BRCA1/2 genes. Responses to multiple lines of chemotherapy were assessed, as well as the effect of BRCA gene mutations on progression-free survival (PFS) and overall survival (OS).ResultsPathogenic BRCA1/2 mutations were found in 21.15% of the patients. BRCA1 mutations represented 68.2% of the total mutations. Two novel BRCA1 mutations were identified. Age at diagnosis was a strong predictor of the presence of a pathogenic BRCA1/2 mutation. Patients with a family history of cancer had a higher incidence of BRCA mutations (P=0.005). As high as 72% of the patients with BRCA mutations were diagnosed at advanced stage. High-grade serous tumors have a higher incidence of pathogenic mutation (P=0.07). Response to neoadjuvant chemotherapy was high (93.9%). All patients underwent surgery which was optimal in 73.1% of the patients. As high as 85.6% of the patients received adjuvant chemotherapy. Relapse rate was 45.2%. Visceral metastasis was more often in BRCA carriers (P=0.01). Patients carrying pathogenic BRCA1/2 mutations had a longer median PFS of 42.43 months (95% CI 32.04–52.83) compared to 22.24 months (95% CI 14.83–29.58) for non-carriers (P=0.08). OS was 64.32 months (95% CI 38.09–90.06) for BRCA mutation patients versus 56.63 months (95% CI 50.05–63.21) (P=0.04) for non-carriers. In multivariate analysis, early stage at diagnosis and optimal debulking were the only independent predictors of better PFS and OS.ConclusionWe documented a number of pathogenic BRCA1 and 2 mutations in this patients cohort; two novel mutations were detected. BRCA status seemed to affect survival in ovarian cancer patients.

Association between family history, mutation locations, and prevalence of BRCA1 or 2 mutations in ovarian cancer patients.

We investigated the prevalence of germline BRCA mutations in a population‐based cohort of Austrian women diagnosed with ovarian cancer and its association with family history of cancer. We prospectively collected family pedigrees of 443 Austrian ovarian cancer patients who had been tested for the presence of a germline BRCA or 2 mutations and correlated the familial breast and ovarian cancer burden with the prevalence of BRCA mutations and disease onset. The probability of carrying a gBRCA mutation in patients without family history of cancer is 14% (95% CI 9%‐22%), as opposed to 45% (95% CI 31%‐59%) of patients with at least one family member with ovarian cancer, and 47% (95% CI 40%‐54%) if other relatives have developed breast cancer. If both breast and ovarian cancer are diagnosed in the family, the probability of carrying a germline BRCA1 or 2 mutations is 60% (95% CI 50%‐68%). germline BRCA1 or mutations in families with ovarian cancer only are commonly located in the Ovarian Cancer Cluster Regions when compared to families with both breast and ovarian cancer (P = 0.001, and P = 0.020, respectively). While gBRCA mutation carriers with ovarian cancer do not have a significantly different age at onset than patients with a family history of cancer, gBRCA1 carriers in general have an earlier onset than gBRCA2 carriers (P = 0.002) and patients without a mutation (P = 0.006). The rate of germline BRCA1 or 2 mutations in ovarian cancer patients without a family history or breast or ovarian cancer is low. However, in women with additional family members affected, the prevalence is considerably higher than previously reported.

LBA5 - Mainstreaming genetic counselling for genetic testing of BRCA1 and BRCA2 in ovarian cancer patients in Malaysia (MaGiC study)

Abstract Background Identification of germline BRCA mutations in ovarian cancer patients allows clinicians to make treatment decisions and manage risks. Due to the lack of genetic counsellors, clinical geneticists and awareness among clinicians, there is minimal testing regionally. Mainstreaming genetic counselling may improve this access. This study aims to determine the prevalence of BRCA mutations, feasibility of mainstreaming and the psychosocial impact of genetic testing in Malaysian ovarian cancer patients. Methods This study aimed to recruit 800 patients with non-mucinous epithelial ovarian, peritoneal or fallopian tube cancer regardless of age or family history, via mainstreaming or referral to genetic services. Genetic Counselling Satisfaction Scale (GCSS), Decisional Conflict Scale (DCS), Psychosocial Aspect of Hereditary Cancer (PAHC) and Cancer Worry Scale (CWS) were used to measure the feasibility and psychosocial outcomes. Mainstreaming clinicians provided feedback through a survey. Results 23/28 mainstreaming sites remained active and 47/68 trained clinicians continued to provide genetic counselling. 800 patients have been recruited, 690 via mainstreaming and 110 via genetics route. 790 have been tested, 110 (13.9%) have pathogenic variants, 90 (11.4%) VUS and 590 (74.7%) negative. Majority of the patients were satisfied with their pre-test GC (mean: 23.9; SD: 2.7) and post result disclosure (mean: 24.6; SD: 2.6). Large majority (86%) did not feel conflicted. Patients with no issues in 5 out of 6 PAHC domains increased from 46% at pre-test GC to 62% at result disclosure. However, only 16% of participants showed high distress level after result disclosure, 22% of these were carriers. Generally, there were no significant differences between mainstreaming and genetics pathways. 36/47 (80%) of clinician are confident to counsel patients and want to integrate BRCA testing into their clinical practice. Conclusions BRCA prevalence for ovarian cancer in Malaysia is 13.9%. There is satisfaction in counselling services, acceptable psychosocial impact with no significant differences between mainstreaming and genetics pathway. Mainstreaming may improve access to genetic testing in Malaysia. Legal entity responsible for the study Cancer Research Malaysia. Funding AstraZeneca. Disclosure S.Y.Y. Yoon: Research grant / Funding (institution), Investigator Initiated Research Grant: AstraZeneca. A. George: Research grant / Funding (institution): Wellcome Trust; Honoraria (institution): AstraZeneca; Honoraria (institution): Roche; Honoraria (institution): Tesaro.

Multigene panel testing increases the detection of clinically actionable mutations in ovarian cancer patients

Objective: Beyond BRCA1/2, a number of genes are associated with an increased risk for ovarian and/or other cancers. The majority of these genes have associated National Comprehensive Cancer Network (NCCN) guidelines for cancer risk management. We aimed to determine the frequency of pathogenic germline mutations in genes, other than BRCA1/2, among women with ovarian cancer undergoing multigene panel testing (MGPT).

LBA4_PR - Mainstreaming genetic counselling for genetic testing of BRCA1 and BRCA2 in ovarian cancer patients in Malaysia (MaGiC study)

Background: Identifying germline BRCA mutations in ovarian cancer patients is important for the medical management of patients and risk management for relatives. In Asia, genetic services are relatively inaccessible mainly due to the lack of genetic counselling. Mainstreaming improved access to cancer genetic testing in United Kingdom, hence its feasibility is being tested in Malaysia. The MaGiC Study aims to (a) determine the prevalence of germline BRCA1/BRCA2 mutations in a population-based cohort (b) determine the feasibility of mainstreaming genetic counselling and testing and (c) investigate the psychosocial impact of genetic testing in Malaysia Methods: This is a prospective observational study of 800 patients with non-mucinous ovarian cancer irrespective of family history, recruited via a mainstreaming pathway where patients are counselled by non-genetic clinicians or the traditional genetics pathway. Phone interviews are conducted after pre-test counselling and result disclosure to study feasibility and psychosocial impact using Genetic Counselling Satisfaction Scale (GCSS), Decisional Conflict Scale (DCS), Psychosocial Aspect of Hereditary Cancer (PAHC) and Cancer Worry Scale (CWS). Results: 70 clinicians from 29 sites nationwide recruited 248 patients. 208 patients were tested. 27 (13%) pathogenic mutations, 33 (16%) variants of uncertain significance and 148 (71%) negatives were identified. Analysis of 160 patients showed that GCSS, DCS, PAHC and CWS were reliable. Most patients were satisfied with their counselling experience, not conflicted in making decision, felt informed about their choices. PAHC showed that 79% patients at pre-test and 69% at post test had a problem mainly on ‘living with cancer’. Distress Thermometer showed 41 of 158 (26%) patients may require psychosocial support at pre-test but reduced to 17% after result disclosure. For CWS, 64 of 156 (41%) patients at pre and 26 of 64 (41%) at post-test reported to have frequent concerns about getting cancer again. Conclusions: Mainstreaming may improve access to cancer genetic testing regionally. Interim results from this study will demonstrate the challenges and the feasibility of introducing mainstreaming cancer genetics in Malaysia. Legal entity responsible for the study: Cancer Research Malaysia Funding: Cancer Research Malaysia and Astra Zeneca Disclosure: S.Y. Yoon: This study is funded by Cancer Research Malaysia and with an Investigator Initiated Grant from Astra Zeneca. Ms Yoon has received honorarium as invited speaker by Astra Zeneca. N. Rahman: Non-executive director of Astra-Zeneca. M.K. Thong, S.H. Teo: Honorarium as speaker from Astra Zeneca. Y.L. Woo: Honorarium as a speaker by Astra Zeneca. All other authors have declared no conflicts of interest.

BRCA1 and BRCA2 mutations in ovarian cancer patients from China: Association of ethnic-specific mutations in BRCA1 with an increased risk of ovarian cancer.

1583 Background: BRCA1/2 are cancer predisposition genes involved in hereditary breast and ovarian cancer (HBOC). Mutation carriers display an increased sensitivity to inhibitors of poly (ADP-ribose) polymerase (PARP). Despite a number of small-size hospital-based studies being previously reported, there is not yet, precise data of BRCA1/2mutations among Chinese epithelial ovarian cancer (EOC) pts. Methods: We performed a multicenter cohort study including 916 unselected consecutive EOC pts from eastern China, to screen for BRCA1/2mutations using the next-generation sequencing approach. Results: 153 EOC pts were found to carry pathogenic germline mutations in BRCA1/2, accounting for an overall mutation incidence of 16.7% with the predominance in BRCA1 (13.1%) compared with BRCA2 (3.9%). We identified 53 novel pathogenic mutations, among which the c.283_286delCTTG and the c.4573C&gt;T of BRCA1 were both found in two unrelated patients. More importantly, the most common mutation, c.5470_5477del8 was most likely to be Chinese population-related without an apparent founder origin. This hot-spot mutation was presumably associated with an increased risk of EOC. Taken together, germline BRCA1/2mutations were common in Chinese EOC pts with distinct mutational spectrum compared to Western populations. Conclusions: Our study contributes to the current understanding of BRCA1/2 mutation prevalence worldwide. We recommend BRCA1/2genetic testing to all Chinese women diagnosed with EOC in order to identify HBOC families, to provide genetic counselling and clinical management for at-risk relatives. Mutation carriers may also benefit from PARP-targeted therapies. [Table: see text]

BRCA1 and BRCA2 mutations in ovarian cancer patients from China: ethnic-related mutations in BRCA1 associated with an increased risk of ovarian cancer.

BRCA1/2 are cancer predisposition genes involved in hereditary breast and ovarian cancer (HBOC). Mutation carriers display an increased sensitivity to inhibitors of poly(ADP-ribose) polymerase (PARP). Despite a number of small-size hospital-based studies being previously reported, there is not yet, to our knowledge, precise data of BRCA1/2 mutations among Chinese ovarian cancer patients. We performed a multicenter cohort study including 916 unselected consecutive epithelial ovarian cancer (EOC) patients from eastern China to screen for BRCA1/2 mutations using the next-generation sequencing approach. A total of 153 EOC patients were found to carry pathogenic germline mutations in BRCA1/2, accounting for an overall mutation incidence of 16.7% with the predominance in BRCA1 (13.1%) compared with BRCA2 (3.9%). We identified 53 novel pathogenic mutations, among which the c.283_286delCTTG and the c.4573C > T of BRCA1 were both found in two unrelated patients. More importantly, the most common mutation found in this study, c.5470_5477del8 was most likely to be Chinese population-related without an apparent founder origin. This hot-spot mutation was presumably associated with an increased risk of ovarian cancer. Taken together, germline BRCA1/2 mutations were common in Chinese EOC patients with distinct mutational spectrum compared to Western populations. Our study contributes to the current understanding of BRCA1/2 mutation prevalence worldwide. We recommend BRCA1/2 genetic testing to all Chinese women diagnosed with EOC to identify HBOC families, to provide genetic counseling and clinical management for at-risk relatives. Mutation carriers may also benefit from PARP-targeted therapies.

A targeted analysis identifies a high frequency of BRCA1 and BRCA2 mutation carriers in women with ovarian cancer from a founder population.

BackgroundThe frequency of BRCA1 and BRCA2 mutations in ovarian cancer patients varies depending on histological subtype and population investigated. The six most commonly recurring BRCA1 and BRCA2 mutations previously identified in a founder French Canadian population were investigated in 439 histologically defined ovarian, fallopian tube and primary peritoneal cancer cases that were ascertained at one hospital servicing French Canadians. To further assess the frequency of BRCA1/BRCA2 mutations, a defined subgroup of 116 cases were investigated for all mutations previously reported in this population.MethodsA PCR-based assay was used to screen 439 ovarian, fallopian tube or extra-ovarian cancers comprised of serous, high grade endometrioid and mixed cell adenocarcinomas with serous components for specific BRCA1: C4446T and 2953delGTAinsC and BRCA2: 8765delAG, G6085T, 3398del5 and E3002K mutations. A multiplex bead-array-based Luminex assay was used to evaluate 19 specific mutations that have ever been reported in French Canadians, which included the six mutations assayed by PCR, in 116 cases representing all women ascertained within a defined 3-year window.ResultsA targeted analysis of six mutations identified 34/439 (7.7%) mutation carriers and at least two mutation carriers for each mutation screened were found. The BRCA1:C4446T mutation was the most frequently identified variant (15/34, 44.1%) among mutation-positive cases. The expanded mutation screen that also included 13 additional variants identified 19/116 (16.4%) mutation carriers, where C4446T was the most common variant (8/19, 42.1%) identified among mutation-positive carriers in this subgroup. Mutations were identified in women with serous, endometrioid, mixed cell, and undifferentiated adenocarcinomas. Within this subgroup there were 73 high-grade (G3) serous ovarian carcinomas, the most common subtype, with mutations identified in 19.2% (n = 14) serous cases.ConclusionsOur results reaffirm that specific BRCA1 and BRCA2 mutations found previously to recur in French Canadian breast cancer and breast-ovarian cancer families, also recur in women with ovarian cancer not selected for family history of cancer. The high frequency of mutation carriers rationalizes genetic testing of ovarian cancer patients in this demographically defined population.Electronic supplementary materialThe online version of this article (doi:10.1186/s13048-015-0124-8) contains supplementary material, which is available to authorized users.

Abstract B30: Induction of BRCAness phenotype by curcumin and hydroxyurea in WT BRCA ovarian cancer cells

Abstract The field of ovarian cancer therapeutics has recently made a substantial leap forward with the discovery that ovarian cancers with a BRCA mutation can be exploited with “synthetic lethality”. This phenomenon takes advantage of the role of PARP (poly-ADP ribosyl-polymerase), which compensates for the missing or low BRCA protein in these patients with a BRCA germline mutation. PARP activity is essential for these patients to maintain a normal genotype, albeit not completely normal since they developed ovarian cancer mainly because of their BRCA mutation. Thus, in the 5 major mechanisms of DNA repair PARP compensates for missing BRCA1 or BRCA2 in homologous repair, base excision repair and nuclear excision repair. In these patients, if PARP is inhibited, then massive DNA mutations accumulate and are not repaired because PARP ribosylates the areas of cut or damaged DNA to mark the areas needed for repair to occur. The concept of inhibiting PARP in these BRCA mutated tumors is an elegant idea. The caveat here is the BRCA mutant genotype where there is a germline, inactivating mutation in one allele and LOH or somatic mutation of the other allele only occurs in 5-10% of the cancer population. Thus, could there be a way to apply this concept of synthetic lethality to the other 90-95% of the cancer population without BRCA mutations in ovarian cancer patients? We tried to answer that question by testing different agent which could modulate gene expression, and found that curcumin and hydroxyurea, at low concentrations, could sometimes increase the cytotoxicity of alkylating agents which suggests a mechanism involving DNA repair. Four (ovcar-8, TOV21G, PA-1 and UACC1598) out of nine pretreated ovarian cancer cell lines with hydroxyurea or curcumin has synergistically increase apoptosis induced by PARP inhibitor - Olaparid. In the tested Ovcar-8 cancer cell line, Curcumin (5 μM) alone induced 10% apoptosis, hydroxyurea (100 μM) alone induced 17% apoptosis, Olaparid (2.5 μM) alone induced 7% cell death. Curcumin and Olaparid together induce 45% cell death (165% increase), hydroxurea and Olaparid together induced 40% (67% increase). Both curcumin and hydroxyurea decrease BRCA1 and BRCA2 mRNA and protein level in the tested sensitive cell lines. Further study, we found that curcumin prompt epigenetically BRCA1,2 promoter methylation and lead to gene expression shutting down whereas hydroxyurea induced miRNA hsa-miR-17-5p and hsa-miR-182-5p and lead BRCA1 mRNA degradation. This finding suggest If wt BRCA patients with ovarian cancer could be induced into BRCAness, then perhaps a larger percentage than 5-10% could benefit from PARP inhibitors and DNA crosslinkers like mitomycin C. Citation Format: Yuehua Mao, Richard D Dinnen, Robert L Fine. Induction of BRCAness phenotype by curcumin and hydroxyurea in WT BRCA ovarian cancer cells. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B30.

Three per cent of Norwegian Ovarian Cancers are caused by BRCA1 1675delA or 1135insA

Our aim was to determine the prevalence of two Norwegian BRCA1 founder mutations in ovarian cancer patients, to identify carriers and their families for medical follow-up, and to study histopathological factors. Of a cohort of 727 ovarian cancer patients, 615 gave informed consent to testing. 2.9% (18/615) of the tested patients were found to be carriers of BRCA1 1675delA ( n=13) or 1135insA ( n=5). The total frequency of the mutations was 4.7% (8/171) in patients below 50 years of age, and zero (0/144) in patients above 70 years of age. In patients below 70 years of age, the frequency of 1675delA and 1135insA mutations was 2.8% and 1.0%, respectively. Out of 13 patients with 1675delA mutation, 4 had breast cancer. 14/16 (87.5%) families fulfilled clinical criteria for familial breast-ovarian cancer. Median age of onset of ovarian and breast cancer was 51 years and 37 years, respectively. Mutation carriers tended to have tumours with unfavourable prognostic factors. This is, to our knowledge, the highest reported frequency of founder mutations in a national ovarian cancer cohort (less than in the Ashkenazis). It seems justified to offer such testing to ovarian cancer patients below 70 years of age in Norway, identify their risk of breast cancer and offer medical follow-up to the families.