KRAS MUTANTS, A KEY ONCOGENE IN NON-SMALL CELL LUNG CANCER

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Date

2024-04-26

Authors

Samarkhanova, Diana

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Publisher

Nazarbayev University School of Medicine

Abstract

Cancer, a disease with a high risk of mortality, continues to pose a significant global health challenge, affecting millions of individuals every year. Non-small cell lung cancer (NSCLC), which accounts for 80–85% of lung cancer cases, is one of the most prevalent cancer types (Xie et al., 2021). The Kirsten rat sarcoma viral oncogene homologue (KRAS) is one of the most commonly mutated genes in NSCLC. KRAS encodes a protein that has both active and inactive states. In cancer, this protein remains persistently active, leading to angiogenesis and tumorigenesis (Matsuo et al., 2009). More than 80% of KRAS mutations are found at codon 12, with the most prevalent mutations being KRAS G12C (40%), KRAS G12V (18–21%), and KRAS G12D (17–18%), among others (Xie et al., 2021). Current research on G12C inhibitors has shown promising cancer therapy candidates such as sotorasib and adagrasib (Kwan et al., 2022). However, acquired resistance to inhibitors of the G12C mutation has been observed over time (Blaquier et al., 2021). Therefore, it is necessary to generate KRAS mutants that can be used for drug screening to analyze the efficacy of drugs and resistance mechanisms. The purpose of this project is to use CRISPR/Cas9-mediated introduction of mutations to develop KRAS mutant models with G12C, G12V and G12D mutation in the H1299 cell line. KRAS mutants further can be used to screen new drug candidates for the KRAS-dependent NSCLC. The motivation behind this effort is the generating cell lines with endogenous mutations by homology-directed repair (HDR) in the KRAS second exon that will mimic natural conditions and show more physiologically relevant data, which is necessary to analyze the effectiveness of different treatments. In summary, this project tested the efficacy of generating knockout of the KRAS gene and the introduction of G12C, G12V and G12D mutations to the second exon of KRAS in the H1299 cell line using CRISPR/Cas9 system. The results of the research project showed the low efficacy of HDR to generate endogenous mutations and the successful application of CRISPR/Cas9 to generate KRAS knockout clones.

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Keywords

Type of access: Restricted, Non-small cell lung cancer, NSCLC, KRAS mutations, KRAS G12C, KRAS G12V, KRAS G12D, CRISPR/Cas9, homology-directed repair, KRAS mutants, drug screening

Citation

Samarkhanova, D. (2024). KRAS mutants, a key oncogene in non-small cell lung cancer. Nazarbayev University School of Medicine