MSE PhD Prospectus Defense: Yuyang Gu

MSE PhD Prospectus Defense: Yuyang Gu TITLE: m5C-modified self-amplifying RNA for in vivo generation of logic-gated CAR-T cells with improved safety and efficacy ADVISOR: Mark Grinstaff BME, MSE, Chemistry COMMITTEE: Wilson Wong BME, Alexander Green BME, Ian Rifkin Medicine, Virology, Immunology & Microbiology ABSTRACT: Chimeric antigen receptor T (CAR T) cell therapy is transforming the treatment landscape of hematological malignancies. However, manufacturing with integrating viral vectors is costly, slow, and carries risks including insertional mutagenesis, prolonged B cell aplasia, and other long-term toxicities. Expression of CAR with mRNA can reduce cost, manufacturing timelines, and improve safety. However, the short-lived expression necessitates frequent repeat dosing. Here, I propose to leverage single-stranded, 5-methylcytidine (m5C)-modified saRNA delivered via targeted lipid nanoparticles to enable the in situ generation of logic-gated CAR T cells with prolonged CAR expression relative to conventional mRNA and programmable antigen-recognition capabilities, ultimately enhancing the safety, efficacy, and cost-effectiveness of CAR T cell therapy. Preliminary studies demonstrate that ex vivo engineering of primary human T cells with m5C-modified saRNA via electroporation prolongs CAR expression, enables durable tumor killing in vitro and in vivo, and supports sophisticated CAR designs such as OR- and AND-gated circuits.