Mechanistic insights into c-Met rs368750834 mutation and a bifunctional CAR-T strategy for serous ovarian carcinoma

The c-Met gene encodes a protein known as the hepatocyte growth factor receptor (HGFR), which is frequently called the c-Met receptor. This gene and its encoded receptor play pivotal roles in normal physiological processes as well as in the initiation, progression, and metastasis of various diseases, particularly cancer. However, the precise molecular mechanisms underlying the missense mutation at residue 110 (valine to isoleucine) within this gene remain unexplored. This mutation occurs hypothesized to impact protein stability; yet, how it alters the three-dimensional structure, dynamic behavior, and functional properties of c-Met is unclear. In this study, we innovatively integrate high-precision structure prediction, protein model-based molecular dynamics simulations, and AI-driven thermodynamic stability analyses to systematically elucidate the molecular mechanism by which the residue 110 mutation disrupts protein structural integrity, modulates conformational dynamics, and ultimately impairs receptor function. Utilizing c-Met as a tumor-associated antigen, we developed a PD-1 antibody-secreting c-Met-targeted CAR-T cell and evaluated its cytotoxic efficacy against the serous ovarian cancer cell line SKOV-3 both in vitro and in vivo. This study reveals that mutation at position 110 of c-Met induces protein conformational reprogramming, transforming it from a stable, compact structural state into a more loosely organised and dynamically flexible conformation. Experimental evidence confirms that this structural alteration leads to significantly elevated protein expression levels of the V110I mutant in ovarian cancer cells compared to the wild-type, functionally demonstrating the substantive impact of altered conformational stability on protein metabolism. Building upon the identification of c-Met protein as a potential therapeutic target in ovarian cancer, we developed c-Met CAR-T cells capable of secreting anti-PD-1 antibodies. These cells exhibited markedly enhanced cytotoxicity against tumour cells overexpressing c-Met, accompanied by increased release of key effector cytokines such as interferon-γ. This research framework offers a precise, synergistic therapeutic strategy to overcome limitations in CAR-T therapy response within serous ovarian carcinoma.