Next-generation sequencing uncovers crucial mutated genes and potential therapeutic targets in ovarian cancer patients.

Abstract

Objectives: Ovarian cancer is a highly lethal gynecological malignancy, often diagnosed late, resulting in high mortality. While BRCA1 and BRCA2 mutations are known risk factors, the broader genetic landscape needs comprehensive profiling to identify additional diagnostic markers or therapeutic targets. The current study aims to explore the genetic landscape of various cancer-susceptible genes in ovarian cancer patients.

Methods: The genetic landscape of ovarian cancer was investigated by analyzing 27 genes via next-generation sequencing (NGS) in 50 ovarian cancer patients.

Results: Mutations were detected in four genes: Breast Cancer 1 (BRCA1) (62%), Cyclin-Dependent Kinase 4 (CDK4) (58%), MutS Homolog 2 (MSH2) (48%), and Phosphatase and Tensin Homolog (PTEN) (22%). Pathogenic mutations were identified in BRCA1 (p.Tyr1853Ter and p.Gln1848Ter), CDK4 (p.Arg24His), and PTEN (p.Tyr29Ter), occurring in 11 patients. Interestingly, these pathogenic mutations were absent in The Cancer Genome Atlas (TCGA) dataset and the gnomAD for the Asian population, suggesting their unique presence in the Pakistani cohort. Functional assays revealed that these mutations significantly reduced the mRNA and protein expression levels of BRCA1, CDK4, and PTEN, as demonstrated by Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) and Immunohistochemistry (IHC) analyses. Receiver Operating Characteristic (ROC) curve analysis confirmed the potential of these genes as biomarkers, with downregulated expression accurately distinguishing between normal and cancerous tissues. Structural validation of mutated proteins using Ramachandran plots and Protein Structure Analysis (ProSA-web) analysis confirmed the stability of the mutations. Drug prediction and molecular docking identified Resveratrol as a potential therapeutic agent, indicating strong binding affinities with BRCA1, CDK4, and PTEN proteins.

Conclusion: These findings provide novel insights into the genetic underpinnings of ovarian cancer in the Pakistani population and suggest potential targets for therapeutic intervention.