Clinico-pathological correlation of homologous recombination status in epithelial ovarian cancer: Surgeon’s perspective

Background: TCGA data using expensive multi-modality diagnostic platforms have shown that 50% epithelial ovarian cancers (EOCs) are estimated to be homologous recombination (HR) deficient (HRD). We developed a functional assay for HR using gamma H2AX-Rad51 immunofluoresence.[1] Methods: Primary cultures were developed in 50 consecutive EOCs from ascetic fluid and HR assay was performed. Results: 50% patients were HRD based on the functional assay and show improved ex-vivo chemosensitivity to PARP inhibitor (PARPi) (PPV = 92%, NPV = 100%). HRD patients showed improved platinum sensitivity (53.8% vs 16.7%), survival (12 month OS - 41.7% vs. 11.5%) and optimal cytoreduction (80% vs. 62%) rates compared to HR competent (HRC) tumours which are less responsive and represent an unmet clinical need. Conclusions: Personalised surgical and chemotherapeutic strategies may be developed for HR stratified EOCs. Primary surgery may be the preferred approach in HRC due to poor chemoresponse; surgical expertise/environment should be optimised to ensure optimal surgical outcome. Intra-operative hyperthermic treatment and selective HR inhibitors may improve subsequent chemoresponse in HRC and are currently being investigated.

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Functional Ex Vivo Assay to Select Homologous Recombination–Deficient Breast Tumors for PARP Inhibitor Treatment

Clinical Cancer Research ◽

10.1158/1078-0432.ccr-14-0571 ◽

2014 ◽

Vol 20 (18) ◽

pp. 4816-4826 ◽

Cited By ~ 86

Author(s):

Kishan A.T. Naipal ◽

Nicole S. Verkaik ◽

Najim Ameziane ◽

Carolien H.M. van Deurzen ◽

Petra ter Brugge ◽

...

Keyword(s):

Homologous Recombination ◽

Ex Vivo ◽

Parp Inhibitor ◽

Breast Tumors ◽

Ex Vivo Assay ◽

Inhibitor Treatment

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Development of a Functional Assay for Homologous Recombination Status in Primary Cultures of Epithelial Ovarian Tumor and Correlation with Sensitivity to Poly(ADP-Ribose) Polymerase Inhibitors

Clinical Cancer Research ◽

10.1158/1078-0432.ccr-09-2758 ◽

2010 ◽

Vol 16 (8) ◽

pp. 2344-2351 ◽

Cited By ~ 182

Author(s):

Asima Mukhopadhyay ◽

Ahmed Elattar ◽

Aiste Cerbinskaite ◽

Sarah J. Wilkinson ◽

Yvette Drew ◽

...

Keyword(s):

Homologous Recombination ◽

Ovarian Tumor ◽

Primary Cultures ◽

Functional Assay ◽

Epithelial Ovarian Tumor ◽

Polymerase Inhibitors

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Advanced Ovarian Cancer Displays Functional Intratumor Heterogeneity That Correlates to Ex Vivo Drug Sensitivity

International Journal of Gynecological Cancer ◽

10.1097/igc.0000000000000745 ◽

2016 ◽

Vol 26 (6) ◽

pp. 1004-1011 ◽

Cited By ~ 6

Author(s):

Rachel L. O’Donnell ◽

Angelika Kaufmann ◽

Laura Woodhouse ◽

Aiste McCormick ◽

Paul A. Cross ◽

...

Keyword(s):

Ovarian Cancer ◽

Homologous Recombination ◽

Epithelial Ovarian Cancer ◽

Solid Tumor ◽

Ex Vivo ◽

Primary Cultures ◽

Advanced Ovarian Cancer ◽

Antigen Expression ◽

Intratumor Heterogeneity ◽

Protein Markers

IntroductionEpithelial ovarian cancer is recognized to be heterogeneous but is currently treated with a single treatment strategy. Successful patient stratification of emerging chemotherapy agents is dependent upon the availability of reliable biomarkers indicative of the entire tumor.AimThe aim of this study was to evaluate intertumor and intratumor heterogeneity within a series of epithelial ovarian cancer using homologous recombination (HR) DNA repair status.MethodsPrimary cultures generated from ascites and solid tumor from multiple intra-abdominal sites were characterized by their morphology and expression of protein markers. Results were compared with Formalin fixed paraffin embedded tissue pathology.Homologous recombination function was determined by quantification of nuclear Rad51 foci. Growth inhibition (sulforhodamine B) assays were used to calculate the GI50 for cisplatin and rucaparib.ResultsAscites with matched solid tumor were cultured from 25 patients.Concordance in functional HR status between ascites and solid tumor subcultures was seen in only 13 (52%) of 25 patients. Heterogeneity in HR status was seen even in patients with homogeneous histological subtype. Homologous recombination defective cultures were significantly more sensitive to cisplatin and rucaparib.Additionally, intertumor and intratumor heterogeneity was seen between the expression of epithelial and ovarian markers (EpCAM, cytokeratin, CA125, MOC-31, and vimentin). There was no relationship between heterogeneity of HR functional status and antigen expression.ConclusionsIntertumor and intratumor functional HR heterogeneity exists that cannot be detected using histological classification. This has implications for biomarker-directed treatment.

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Exploring the Frequency of Homologous Recombination DNA Repair Dysfunction in Multiple Cancer Types

Cancers ◽

10.3390/cancers11030354 ◽

2019 ◽

Vol 11 (3) ◽

pp. 354 ◽

Cited By ~ 6

Author(s):

Lucy Gentles ◽

Bojidar Goranov ◽

Elizabeth Matheson ◽

Ashleigh Herriott ◽

Angelika Kaufmann ◽

...

Keyword(s):

Overall Survival ◽

Dna Repair ◽

Homologous Recombination ◽

Ex Vivo ◽

Primary Cultures ◽

Ovarian Cancer Cells ◽

Continuous Exposure ◽

Multiple Cancer ◽

Brca Mutations ◽

Platinum Chemotherapy

Dysfunctional homologous recombination DNA repair (HRR), frequently due to BRCA mutations, is a determinant of sensitivity to platinum chemotherapy and poly(ADP-ribose) polymerase inhibitors (PARPi). In cultures of ovarian cancer cells, we have previously shown that HRR function, based upon RAD51 foci quantification, correlated with growth inhibition ex vivo induced by rucaparib (a PARPi) and 12-month survival following platinum chemotherapy. The aim of this study was to determine the feasibility of measuring HRR dysfunction (HRD) in other tumours, in order to estimate the frequency and hence wider potential of PARPi. A total of 24 cultures were established from ascites sampled from 27 patients with colorectal, upper gastrointestinal, pancreatic, hepatobiliary, breast, mesothelioma, and non-epithelial ovarian cancers; 8 were HRD. Cell growth following continuous exposure to 10 μM of rucaparib was lower in HRD cultures compared to HRR-competent (HRC) cultures. Overall survival in the 10 patients who received platinum-based therapy was marginally higher in the 3 with HRD ascites (median overall survival of 17 months, range 10 to 90) compared to the 7 patients with HRC ascites (nine months, range 1 to 55). HRR functional assessment in primary cultures, from several tumour types, revealed that a third are HRD, justifying the further exploration of PARPi therapy in a broader range of tumours.

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Abstract CT161: A homologous recombination repair (HRR) functional assay to stratify patients with metastatic prostate cancer for PARP inhibitor treatment in the TOPARP-B clinical trial

10.1158/1538-7445.am2021-ct161 ◽

2021 ◽

Author(s):

Sara Arce-Gallego ◽

Alba Llop-Guevara ◽

Suzanne Carreira ◽

Nuria Porta ◽

Roberta Fasani ◽

...

Keyword(s):

Prostate Cancer ◽

Clinical Trial ◽

Homologous Recombination ◽

Metastatic Prostate Cancer ◽

Parp Inhibitor ◽

Homologous Recombination Repair ◽

Functional Assay ◽

Recombination Repair ◽

Inhibitor Treatment

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The RAD51-FFPE Test; Calibration of a Functional Homologous Recombination Deficiency Test on Diagnostic Endometrial and Ovarian Tumor Blocks

Cancers ◽

10.3390/cancers13122994 ◽

2021 ◽

Vol 13 (12) ◽

pp. 2994

Author(s):

Lise M. van Wijk ◽

Claire J. H. Kramer ◽

Sylvia Vermeulen ◽

Natalja T. ter Haar ◽

Marthe M. de Jonge ◽

...

Keyword(s):

Homologous Recombination ◽

Ex Vivo ◽

Parp Inhibitor ◽

Attractive Alternative ◽

Fresh Tissue ◽

Rad51 Protein ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Clinical Interest ◽

Formalin Fixed

PARP inhibitor (PARPi) sensitivity is related to tumor-specific defects in homologous recombination (HR). Therefore, there is great clinical interest in tests that can rapidly and reliably identify HR deficiency (HRD). Functional HRD tests determine the actual HR status by using the (dis)ability to accumulate RAD51 protein at sites of DNA damage as read-out. In this study, we further improved and calibrated a previously described RAD51-based functional HRD test on 74 diagnostic formalin-fixed paraffin-embedded (FFPE) specimens (RAD51-FFPE test) from endometrial cancer (EC n = 25) and epithelial ovarian cancer (OC n = 49) patients. We established optimal parameters with regard to RAD51 foci cut-off (≥ 2) and HRD threshold (15%) using matched endometrial and ovarian carcinoma specimens for which HR status had been established using a RAD51-based test that required ex vivo irradiation of fresh tissue (RECAP test). The RAD51-FFPE test detected BRCA deficient tumors with 90% sensitivity and RECAP-HRD tumors with 87% sensitivity, indicating that it is an attractive alternative to DNA-based tests with the potential to be applied in routine diagnostic pathology.

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Faculty Opinions recommendation of 2-Hydroxyglutarate produced by neomorphic IDH mutations suppresses homologous recombination and induces PARP inhibitor sensitivity.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.727261055.793528252 ◽

2017 ◽

Author(s):

Jim Maher

Keyword(s):

Homologous Recombination ◽

Parp Inhibitor ◽

Idh Mutations

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Inhibition of protein N-glycosylation by 2-deoxy-2-fluoro-d-galactose

Biochemical Journal ◽

10.1042/bj2850821 ◽

1992 ◽

Vol 285 (3) ◽

pp. 821-826 ◽

Cited By ~ 3

Author(s):

V Gross ◽

W E Hull ◽

U Berger ◽

T Andus ◽

W Kreisel ◽

...

Keyword(s):

Interleukin 6 ◽

Ex Vivo ◽

Primary Cultures ◽

Blood Monocytes ◽

Galactose Metabolism ◽

Rat Hepatocyte ◽

Peripheral Blood Monocytes ◽

Efficient Inhibitor ◽

Acid Glycoprotein ◽

Alpha 1 Antitrypsin

The effects of 2-deoxy-2-fluoro-D-galactose (dGalF) on N- and O-glycosylation of proteins was studied in rat hepatocyte primary cultures and in human monocytes. In hepatocytes, dGalF at concentrations of 1 mM or higher completely inhibited N-glycosylation of alpha 1-antitrypsin and alpha 1-acid glycoprotein, whereas 4 mM-2-deoxy-D-galactose (dGal) only slightly impaired N-glycosylation. In monocytes, 1 mM- or 4 mM-dGalF blocked N-glycosylation of alpha 1-antitrypsin and of interleukin-6, while O-glycosylation of interleukin-6 remained unaffected. In monocytes, dGal had no effect on protein N-glycosylation. Addition of uridine effectively prevented the UTP deficiency induced by dGalF, but had no effect on the inhibition of protein N-glycosylation by dGalF. Using 19F-n.m.r. spectroscopy, 2-deoxy-2-fluoro-D-galactose 1-phosphate (dGalF-1-P), UDP-dGalF and UDP-dGlcF could be identified as the major metabolites of dGalF in hepatocytes as well as in monocytes. In conclusion, compared with dGal, dGalF is a more efficient inhibitor of protein N-glycosylation. The effect is not caused by the depletion of UTP induced by dGalF, but rather by metabolites of dGalF. dGalF is metabolized not only in hepatocytes but also in peripheral blood monocytes, which can be used for ex vivo studies of disturbances in D-galactose metabolism.

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