Lipid Nanoparticle Delivery of CAR276 mRNA Enables Antigen-Specific Cytotoxicity in T Cells Against Glioblastoma Cells

Background/Objectives: Glioblastoma multiforme (GBM) remains the most aggressive and treatment-refractory primary brain tumor, with current therapeutic modalities providing minimal survival improvement. Chimeric antigen receptor (CAR)-based immunotherapies succeeded in hematologic malignancies but are constrained in solid tumors by tumor heterogeneity, immunosuppressive microenvironment, and delivery challenges. CD276 (B7-H3), a pan-cancer antigen overexpressed in gliomas and correlated with poor patient outcomes, represents a compelling immunotherapeutic target. Methods : Here, we developed a non-viral lipid nanoparticle (LNP)-based platform for transient CAR mRNA delivery to T cells and evaluated its application for generating CD276 (B7-H3)-targeted CAR-T cells against GBM. An in-house-assembled T-mixer incorporating polyether ether ketone (PEEK) tubing was integrated with a dual-syringe-pump system for controlled LNP production. Results : LNPs formulated with clinically relevant ionizable lipids, SM-102 and DLin-MC3-DMA, demonstrated high encapsulation efficiency and effective mRNA delivery. Cell-type-dependent expression kinetics were further characterized following LNP-mediated reporter mRNA delivery in K562, NK, NK92, and T cells, revealing differences in transgene expression across cell populations. LNP-mediated delivery of CD276 CAR mRNA enabled generation of CAR-expressing T cells that exhibited antigen-specific cytotoxicity against CD276-positive LN229 GBM cells in vitro. Conclusions : Collectively, this study demonstrates the feasibility of an adaptable LNP-based platform for non-viral CAR mRNA engineering of T cells and CD276-targeted cytotoxicity against GBM cells in vitro. The combination of transient CAR expression and LNP-mediated mRNA delivery provides a foundation for further optimization and evaluation in preclinical in vivo models.

Abstract from PubMed Central (PMID 42940171, PMC13702138). This entry was added automatically by our daily literature monitor because it matches the topics we follow; read the full paper at the original source.