HITE: HIV Inspired Lipid Nanoparticle Platform for CAR T Cell Engineering.

Chimeric antigen receptor (CAR)-T therapy has led to remarkable advancements in the treatment of hematologic malignancies, encouraging extensive studies on its application to solid tumors and other diseases. However, the production of CAR-T cells is mostly achieved through viral transduction, which results in permanent CAR expression in T cells, potentially leading to unintended adverse effects. Here, we present a lipid nanoparticle (LNP) platform for mRNA delivery to human primary T cells, inspired by the human immunodeficiency virus (HIV) which naturally infects T cells. We perform multiple rounds of screening to sequentially optimize the structure and ratio of ionizable lipid in the base formulation, the ratios of HIV lipid components, and the type and ratio of PEG-lipid for CD3 antibody conjugation. Our HIV envelope-Inspired T cell transfection-Enhancing (HITE) LNP enables efficient generation of CAR-T cells with potent cytotoxic activity against cancer cells in vitro, demonstrating its potential for efficient CAR-T cell production.