By Brooke Martin

Faculty Mentor: Laura Sipe

Abstract

Investigating Methionine Restriction–Induced Pyroptosis as a Mechanism for Enhanced Chemotherapy Response in TNBC
Authors: Brooke Martin, Mallory Thompson, Advisor: Laura Sipe, PhD
Triple-negative breast cancer (TNBC) lacks targetable hormone receptors, making chemotherapy the primary treatment option, therefore investigating ways to make chemotherapy more effective is of clinical need. Previous findings in the lab show methionine restriction, a dietary intervention limiting the amino acid methionine, sensitizes cancer cells to chemotherapy-induced immunogenic cell death, specifically through ATP release in the TNBC cell line EO771. This study investigates how methionine restriction increases chemotherapy-induced ATP release. One potential way is through pyroptosis, where Gasdermin proteins form a pore on the membrane that ATP is released through. Cells were grown in methionine-restricted media at varying concentrations (100%, 40%, 20%, 10%, and 3% methionine) for 72 hours, and gasdermin E (GSDME) gene expression was analyzed by qPCR. Results demonstrated dose-dependent upregulation of GSDME with increasing methionine restriction. Efforts to confirm GSDME protein expression via western blot are ongoing; troubleshooting identified the primary antibody as non-functional, and optimized conditions are being developed for future experiments. These findings suggest methionine restriction may enhance pyroptotic pathways in TNBC, with broader implications for improving chemotherapy efficacy across multiple cancer types.