Prevalence of tumor BRCA1 and BRCA2 dysfunction in unselected patients with ovarian cancer

Objective The therapeutic benefits of poly(ADP-ribose) polymerase inhibitors highlight the need to evaluate BRCA1/2 defects in tubal/ovarian cancer (OC). We sought to determine the pattern and disease characteristics associated with tumor BRCA1/2 mutations and BRCA1 methylation in women with OC. Methods We obtained 111 OC specimens from 2 university hospitals and assessed BRCA1/2 mutations and BRCA1 methylation in tumor DNA. The frequency and pattern of BRCA1/2 defects were examined. Associations between patient/disease characteristics and BRCA1/2 defects were ascertained (Fisher’s exact test). Platinum-free interval (PFI), progression-free survival (PFS), and overall survival (OS) based on the underlying BRCA1/2 defect were determined (Kaplan-Meier analysis [log-rank test]). Results We observed a BRCA1/2 dysfunction rate of 40% (28/70) in high-grade serous tubal/ovarian cancer (HGSC), including 14.3% BRCA1 methylation (n=10), 7.1% BRCA1 mutation (n=5), and 18.6% BRCA2 mutation (n=13). Defects in BRCA1/2 genes were associated with stage III/IV HGSC (BRCA1 methylation: P=0.005 [stage III/IV] and P=0.004 [HGSC]; BRCA1/2 mutation: P=0.03 [stage III/IV] and P<0.001 [HGSC]). Patients with BRCA1/2-mutated cancers showed improved OS (hazard ratio [HR], 0.65; 95% confidence interval [CI], 0.43–0.99; P=0.045) and a trend toward improved PFI (HR, 0.48; 95% CI, 0.22–1.06; P=0.07) and PFS (HR, 0.72; 95% CI, 0.51–1.03; P=0.07). No survival differences were observed between BRCA1-methylated and BRCA1/2 wild-type non-BRCA1-methylated cancers. Conclusion We observed a high tumor BRCA1/2 dysfunction rate in HGSC with a unique predominance of BRCA2 over BRCA1 mutations. While BRCA1/2 mutations conferred survival benefits in OC, no such association was observed with BRCA1 methylation

BRCA1 promoter methylation and clinical outcomes in ovarian cancer: an individual patient data meta-analysis

BACKGROUND BRCA1 methylation has been associated with homologous recombination deficiency, a biomarker of platinum sensitivity. Studies evaluating BRCA1-methylated tubal/ovarian cancer (OC) do not consistently support improved survival following platinum chemotherapy. We examine the characteristics of BRCA1-methylated OC in a meta-analysis of individual participant data. METHODS 2636 participants' data across 15 studies were analyzed. BRCA1-methylated tumors were defined according to their original study. Associations between BRCA1 methylation and clinico-pathological characteristics were evaluated. The effects of methylation on overall survival (OS) and progression-free survival (PFS) were examined using mixed-effects models. All statistical tests were two-sided. RESULTS 430 (16.3%) tumors were BRCA1-methylated. BRCA1 methylation was associated with younger age and advanced-stage high-grade serous OC. There were no survival differences between BRCA1-methylated and non-BRCA1-methylated OC (median PFS = 20.0 vs 18.5 months, HR = 1.01, 95% CI = 0.87-1.16, P=0.98; median OS = 46.6 vs 48.0 months, HR = 1.02, 95% CI = 0.87-1.18, P=0.96). Where BRCA1/2 mutations were evaluated (n = 1248), BRCA1 methylation displayed no survival advantage over BRCA1/2 intact (BRCA1/2 wild type non-BRCA1-methylated) OC. Studies used different methods to define BRCA1 methylation. Where BRCA1 methylation was determined using methylation-specific PCR and gel electrophoresis (n = 834), it was associated with improved survival (PFS: HR = 0.80, 95% CI = 0.66-0.97, P=0.02; OS: HR = 0.80, 95% CI = 0.63-1.00, P=0.05) on mixed-effects modelling. CONCLUSION BRCA1-methylated OC displays similar clinico-pathological features to BRCA1-mutated OC, but is not associated with survival. Heterogeneity within BRCA1 methylation assays influences associations. Refining these assays may better identify cases with silenced BRCA1 function and improved patient outcomes

European experts consensus: BRCA/homologous recombination deficiency testing in first-line ovarian cancer

Background: Homologous recombination repair (HRR) enables fault-free repair of double-stranded DNA breaks. HRR deficiency is predicted to occur in around half of high-grade serous ovarian carcinomas. Ovarian cancers harbouring HRR deficiency typically exhibit sensitivity to poly-ADP ribose polymerase inhibitors (PARPi). Current guidelines recommend a range of approaches for genetic testing to identify predictors of sensitivity to PARPi in ovarian cancer and to identify genetic predisposition. Design: To establish a European-wide consensus for genetic testing (including the genetic care pathway), decision making and clinical management of patients with recently diagnosed advanced ovarian cancer, and the validity of biomarkers to predict the effectiveness of PARPi in the first-line setting. The collaborative European experts’ consensus group consisted of a steering committee (n = 14) and contributors (n = 84). A (modified) Delphi process was used to establish consensus statements based on a systematic literature search, conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines. Results: A consensus was reached on 34 statements amongst 98 caregivers (including oncologists, pathologists, clinical geneticists, genetic researchers, and patient advocates). The statements concentrated on (i) the value of testing for BRCA1/2 mutations and HRR deficiency testing, including when and whom to test; (ii) the importance of developing new and better HRR deficiency tests; (iii) the importance of germline non-BRCA HRR and mismatch repair gene mutations for predicting familial risk, but not for predicting sensitivity to PARPi, in the first-line setting; (iv) who should be able to inform patients about genetic testing, and what training and education should these caregivers receive. Conclusion: These consensus recommendations, from a multidisciplinary panel of experts from across Europe, provide clear guidance on the use of BRCA and HRR deficiency testing for recently diagnosed patients with advanced ovarian cancer

Ovarian cancer molecular pathology.

Peer reviewe

Taxane monotherapy regimens for the treatment of recurrent epithelial ovarian cancer

This is a protocol for a Cochrane Review (Intervention). The objectives are as follows: To assess the efficacy and toxicity of different taxane monotherapy regimens for women with recurrent epithelial ovarian, tubal or primary peritoneal cancer

PO-495 PI3K pathway upregulation mediates acquired resistance to platinum agents and polyadenoribose polymerase inhibitors (PARPi) in BRCA1-methylated ovarian cancer (OC)

Introduction BRCA1-methylated OC (BMOC) are specifically sensitive to platinums and PARPi, though acquired resistance to these agents eventually develops. Elucidating underlying druggable resistance mechanisms is needed to enable novel therapeutic options in BMOC. Material and methods We developed two PARPi resistant (olaparib and talazoparib) and one carboplatin resistant OC cell line models (named OVC8RO, OVC8RT and OVC8RC, respectively) derived from the BRCA1-methylated cell line OVCAR8, following continuous (PARPi) or pulsed (carboplatin) drug exposure. Fold resistance (FR) to the parent drug (as determined by the ratio of the resistant cell line IC50 to the parent cell line IC50) suggested clinically relevant resistance models in OVC8RC (FR=4.80±0.43) and OVC8RO (FR=5.71±0.21). OVC8RT displayed higher level resistance (FR=45.61±11.10). We obtained tissue from 5 matched primary and recurrent (post platinum) BMOC patient tumours. Reverse phase protein array (RPPA) was used to examine differential expression and phosphorylation levels of 63 proteins between parent/resistant cell lines, and primary/matched recurrent tumours. 5 day acid phosphatase cytotoxicity assays were used to determine the IC50 of drugs. Synergy in drug combination assays was determined as per the Chou-Talalay method. Results and discussions Significant increases in PI3K p110a (except in OVC8RT) and AKT S473, along with a significant decrease in PTEN were seen in all resistant cell lines, relative to the parent cell line’s baseline levels (p<0.05), consistent with PI3K pathway upregulation. 3/5 recurrent tumours had increased phosphorylated AKT (T308 or S473), as compared to the corresponding primary BRCA1-methylated tumour. The selective a/δ isoform dominant PI3K inhibitor copanlisib (BAY 80–6946) displayed anti-proliferative effects in both the parent cell line (IC5076.6±7.4 nM) and all resistant cell lines (IC50s 44.0–102.1 nM). In both carboplatin and PARPi-resistant models, combination treatment of copanlisib and the parental drug resulted in synergistic growth inhibition (CI@ ED500.27–0.28) and restored sensitivity to the parent drug. When the parental cells were treated with copanlisib in combination with carboplatin or either PARPi, the observed synergism was markedly less (CI@ED50=0.71–0.85) than that observed in the drug resistant models. Conclusion The addition of copanlisib to carboplatin or PARPi could represent a novel therapeutic strategy in BMOC that has acquired resistance to either carboplatin or PARPi

Comparative selection strategies for development of platinum-drug resistant ovarian cancer cell lines

The development of a drug-resistant cell line in vitro can take from 3 to 18 months. However, little is published on the methodology of this development process. Clinically relevant drug-resistant cell lines are developed by mimicking the conditions cancer patients experience during chemotherapy Doses of drug administered are low, and a pulsed treatment strategy is often used where the cells recover in drug-free media. It is common to have difficulty developing stable clinically relevant drug-resistant cell lines. A comparative selection strategy of multiple cell lines or multiple chemotherapeutic agents mitigates this risk and gives insight into which agents or which clinical phenotypes develops resistance easily. Cell models were chosen to represent different phenotypes of clinical ovarian cancer BRCA1/2 Wildtype (UPN-251, CAOV3), BRCA Mutated (UWB1.289) and BRCA1 Methylated (OVCAR8). Pulsed selection with drug was used to develop resistance to carboplatin and taxol as these drugs are delivered by an I.V bolus. A continuous selection strategy was used to develop resistance to parp inhibitors olaparib and talazoparib as patients take these drugs orally every day. Continuous treatment with parp inhibitors led to stably drug resistant models in all cell lines treated (CAOV3, UWB1.289 and OVCAR8). Resistance to carboplatin was detected in the daughter sublines after 1 - 3 rounds of drug treatment, which was continued to complete 7– 8 rounds, equating to 5-6 months of drug treatment. Alternating the treatment of carboplatin and taxol slowed the development of stable drug resistance compared to treating with single agent (UPN-251 and OVCAR8). The BRCA1 methylated cell line (OVCAR8) was harder to develop into a carboplatin resistant cell line than the other models. In one strategy OVCAR8 models were abandoned as unstably resistant after 7 treatment cycles over 6 months. In a new treatment strategy OVCAR8 developed carboplatin resistance after an extended treatment period 13 cycles over 11 months. BRCA1 methylated patients may therefore benefit from treatment with carboplatin in part due to the slower onset of carboplatin resistance

Taxane monotherapy regimens for the treatment of recurrent epithelial ovarian cancer (Review)

Background Ovarian cancer is the seventh most frequent cancer diagnosis worldwide, and the eighth leading cause of cancer mortality. Epithelial ovarian cancer is the most common kind, accounting for 90% of cases. First-line therapy for women with epithelial ovarian cancer consists of a combination of cytoreductive surgery and platinum and taxane-based chemotherapy. However, more than 50% of women with epithelial ovarian cancer will experience a relapse and require further chemotherapy and at some point develop resistance to platinum-based drugs. Currently, guidance on the use of most chemotherapy drugs, including taxanes, is unclear for women whose epithelial ovarian cancer has recurred. Paclitaxel, topotecan, pegylated liposomal doxorubicin hydrochloride, trabectedin and gemcitabine are all licensed for use in the UK at the discretion of clinicians, following discussion with the women as to potential adverse effects. Taxanes can be given in once-weekly regimens (at a lower dose) or three-weekly regimens (at a higher dose), which may have differences in the severity of side effects and effectiveness. As relapsed disease suggests incurable disease, it is all the more important to consider side effects and the impact of treatment schedules, as well as quality of life, and not only the life-prolonging effects of treatment. Objectives To assess the efficacy and toxicity of different taxane monotherapy regimens for women with recurrent epithelial ovarian, tubal or primary peritoneal cancer. Search methods We searched CENTRAL, MEDLINE and Embase, up to 22 March 2022. Other related databases and trial registries were searched as well as grey literature and no additional studies were identified. A total of 1500 records were identified. Selection criteria We included randomised controlled trials of taxane monotherapy for adult women diagnosed with recurrent epithelial ovarian, tubal or primary peritoneal cancer, previously treated with platinum-based chemotherapy. We included trials comparing two or more taxane monotherapy regimens. Participants could be experiencing their first recurrence of disease or any line of recurrence. Data collection and analysis Two review authors screened, independently assessed studies, and extracted data from the included studies. The clinical outcomes we examined were overall survival, response rate, progression-free survival, neurotoxicity, neutropenia, alopecia, and quality of life. We performed statistical analyses using fixed-effect and random-effects models following standard Cochrane methodology. We rated the certainty of evidence according to the GRADE approach. Main results Our literature search yielded 1500 records of 1466 studies; no additional studies were identified by searching grey literature or hand searching. We uploaded the search results into Covidence. After the exclusion of 92 duplicates, we screened titles and abstracts of1374 records. Of these, we identified 24 studies for full-text screening. We included four parallel-group randomised controlled trials (RCTs).All trials were multicentred and conducted in a hospital setting. The studies included 981 eligible participants with recurrent epithelial ovarian cancer, tubal or primary peritoneal cancer with a median age ranging between 56 to 62 years of age. All participants had a WHO(World Health Organization) performance status of between 0 to 2. The proportion of participants with serous histology ranged between56% to 85%. Participants included women who had platinum-sensitive (71%) and platinum-resistant (29%) relapse. Some participants were taxane pre-treated (5.6%), whilst the majority were taxane-naive (94.4%). No studies were classified as having a high risk of bias for any of the domains in the Cochrane risk of bias tool. We found that there may be little or no difference in overall survival (OS) between weekly paclitaxel and three-weekly paclitaxel, but the evidence is very uncertain (risk ratio (RR) of 0.94, 95% confidence interval (CI) 0.66 to 1.33, two studies, 263 participants, very low-certainty evidence). Similarly, there may be little or no difference in response rate (RR of 1.07, 95% CI 0.78 to 1.48, two studies, 263 participants, very low-certainty evidence) and progression-free survival (PFS) (RR of 0.83, 95% CI 0.46 to 1.52, two studies, 263 participants, very low-certainty evidence) between weekly and three-weekly paclitaxel, but the evidence is very uncertain. We found differences in the chemotherapy-associated adverse events between the weekly and three-weekly paclitaxel regimens. The weekly paclitaxel regimen may result in a reduction in neutropenia (RR 0.51, 95% 0.27 to 0.95, two studies, 260 participants, low-certainty evidence) and alopecia (RR 0.58, 95% CI 0.46 to 0.73, one study, 205 participants, low-certainty evidence). There may be little or no difference in neurotoxicity, but the evidence was very low-certainty and we cannot exclude an effect (RR 0.53, 95% CI 0.19 to 1.45, two studies, 260 participants).When examining the effect of paclitaxel dosage in the three-weekly regimen, the 250 mg/m2 paclitaxel regimen probably causes more neurotoxicity compared to the 175 mg/m2 regimen (RR 0.41, 95% CI 0.21 to 0.80, one study, 330 participants, moderate-certainty evidence).Quality-of-life data were not extractable from any of the included studies. Authors' conclusions Fewer people may experience neutropenia when given weekly rather than three-weekly paclitaxel (low-certainty evidence), although it may make little or no difference to the risk of developing neurotoxicity (very low-certainty evidence). This is based on the participants receiving lower doses of drug more often. However, our confidence in this result is low and the true effect may be substantially different from the estimate of the effect. Weekly paclitaxel probably reduces the risk of alopecia, although the rates in both arms were high (46%versus 79%) (low-certainty evidence). A change to weekly from three-weekly chemotherapy could be considered to reduce the likelihood of toxicity, as it may have little or no negative impact on response rate (very low-certainty evidence), PFS (very low-certainty evidence) or OS (very low-certainty evidence).Three-weekly paclitaxel, given at a dose of 175 mg/m2 compared to a higher dose, probably reduces the risk of neurotoxicity. We are moderately confident in this result; the true effect is likely to be close to the estimate of the effect, but there is a possibility that it is substantially different. A change to 175 mg/m2 paclitaxel (from a higher dose), if a three-weekly regimen is used, probably has little or no negative impact on PFS or OS (very low-certainty evidence)

Geologic implications of gas hydrates in the offshore of India: Results of the National Gas Hydrate Program Expedition 01

The Indian National Gas Hydrate Program Expedition 01 (NGHP-01) is designed to study the occurrence of gas hydrate along the passive continental margin of the Indian Peninsula and in the Andaman convergent margin, with special emphasis on understanding the geologic and geochemical controls on the occurrence of gas hydrate in these two diverse settings. The NGHP-01 expedition established the presence of gas hydrates in the Krishna–Godavari and Mahanadi Basins, and the Andaman Sea. The expedition discovered in the Krishna–Godavari Basin one of the thickest gas hydrate accumulations ever documented, in the Andaman Sea one of the thickest and deepest gas hydrate stability zones in the world, and established the existence of a fully developed gas hydrate petroleum system in all three basins. The primary goal of NGHP-01 was to conduct scientific ocean drilling/coring, logging, and analytical activities to assess the geologic occurrence, regional context, and characteristics of gas hydrate deposits along the continental margins of India. This was done in order to meet the long-term goal of exploiting gas hydrate as a potential energy resource in a cost effective and safe manner. During its 113.5-day voyage, the D/V JOIDES Resolution cored and/or drilled 39 holes at 21 sites (1 site in Kerala–Konkan, 15 sites in Krishna–Godavari, 4 sites in Mahanadi, and 1 site in the Andaman deep offshore area), penetrated more than 9250 m of sedimentary section, and recovered nearly 2850 m of core. Twelve holes were logged with logging-while-drilling (LWD) tools and an additional 13 holes were wireline logged. The science team utilized extensive on-board laboratory facilities to examine and prepare preliminary reports on the physical properties, geochemistry, and sedimentology of all the data collected prior to the end of the expedition. Samples were also analyzed in additional post-expedition shore-based studies conducted in leading laboratories around the world. One of the specific objectives of this expedition was to test gas hydrate formation models and constrain model parameters, especially those that account for the formation of concentrated gas hydrate accumulations. The necessary data for characterizing the occurrence of in situ gas hydrate, such as interstitial water chlorinities, core-derived gas chemistry, physical and sedimentological properties, thermal images of the recovered cores, and downhole measured logging data (LWD and/or conventional wireline log data), were obtained from most of the drill sites established during NGHP-01. Almost all of the drill sites yielded evidence for the occurrence of gas hydrate; however, the inferred in situ concentration of gas hydrate varied substantially from site to site. For the most part, the interpretation of downhole logging data, core thermal images, interstitial water analyses, and pressure core images from the sites drilled during NGHP-01 indicate that the occurrence of concentrated gas hydrate is mostly associated with the presence of fractures in the sediments, and in some limited cases, by coarser grained (mostly sand-rich) sediments

Geologic implications of gas hydrates in the offshore of India: Krishna–Godavari Basin, Mahanadi Basin, Andaman Sea, Kerala–Konkan Basin

Gas hydrate resource assessments that indicate enormous global volumes of gas present within hydrate accumulations have been one of the primary driving forces behind the growing interest in gas hydrates. Gas hydrate volumetric estimates in recent years have focused on documenting the geologic parameters in the “gas hydrate petroleum system” that control the occurrence of gas hydrates in nature. The primary goals of this report are to review our present understanding of the geologic controls on the occurrence of gas hydrate in the offshore of India and to document the application of the petroleum system approach to the study of gas hydrates. National Gas Hydrate Program of India executed the National Gas Hydrate Program Expedition 01 (NGHP-01) in 2006 in four areas located on the eastern and western margins of the Indian Peninsula and in the Andaman Sea. These areas have experienced very different tectonic and depositional histories. The peninsular margins are passive continental margins resulting from a series of rifting episodes during the breakup and dispersion of Gondwanaland to form the present Indian Ocean. The Andaman Sea is bounded on its western side by a convergent margin where the Indian plate lithosphere is being subducted beneath southeast Asia. NGHP-01 drilled, logged, and/or cored 15 sites (31 holes) in the Krishna–Godavari Basin, 4 sites (5 holes) in the Mahanadi Basin, 1 site (2 holes) in the Andaman Sea, and 1 site (1 hole) in the Kerala–Konkan Basin. Holes were drilled using standard drilling methods for the purpose of logging-while-drilling and dedicated wireline logging; as well as through the use of a variety of standard coring systems and specialized pressure coring systems. NGHP-01 yielded evidence of gas hydrate from downhole log and core data obtained from all the sites in the Krishna–Godavari Basin, the Mahanadi Basin, and in the Andaman Sea. The site drilled in the Kerala–Konkan Basin during NGHP-01 did not yield any evidence of gas hydrate. Most of the downhole log-inferred gas hydrate and core-recovered gas hydrate were characterized as either fracture-filling in clay-dominated sediments or as pore-filling or grain-displacement particles disseminated in both fine- and coarse-grained sediments. Geochemical analyses of gases obtained from sediment cores recovered during NGHP-01 indicated that the gas in most all of the hydrates in the offshore of India is derived from microbial sources; only one site in the Andaman Sea exhibited limited evidence of a thermogenic gas source. The gas hydrate petroleum system concept has been used to effectively characterize the geologic controls on the occurrence of gas hydrates in the offshore of India