Dagher Oula K, Pedard Martin, Bedoya Darel Martinez, Brookens Shawna K, Migliorini Denis, Posey Avery D
Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
Center for Cellular Immunotherapies, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
Mol Ther Nucleic Acids. 2025 Aug 11;36(3):102676. doi: 10.1016/j.omtn.2025.102676. eCollection 2025 Sep 9.
Traditional viral-based chimeric antigen receptor (CAR) T cell therapies have vanquished multiple blood malignancies with decade-long remissions yet struggle against solid tumors. Nonviral engineering of CAR T cells via electroporation or lipid nanoparticle (LNP) delivery of CAR-encoding mRNA results in highly efficient yet transient CAR expression, challenging the adequacy of available preclinical models for mRNA-based CAR T cell evaluation. This study presents a unique three-pronged approach that combines mRNA-based CAR T cells, multi-targeting of glioblastoma (GBM)-associated receptors, and maximal surgical resection as a novel and readily translatable platform for preclinical evaluation of mRNA-based CAR T cells against solid tumors. We performed head-to-head and analyses of mRNA-based CAR T cells generated using different expansion conditions, mRNA delivery methods, or combination approaches. Besides potent cytotoxicity, our findings unveil a therapeutic window of anti-tumor efficacy, as well as robust and durable complete remissions in xenograft mouse models of GBM receiving maximal surgical resection and locoregional injections of multivalent CAR T cells (MVCAR). Such efficacies were significantly better in 5-day expanded versus quiescent T cells. Interestingly, MVCAR T cells were superior to pooled CAR T cells (CARPool) expressing the same CAR scFv combinations in an orthotopic resection model of GBM.
传统的基于病毒的嵌合抗原受体（CAR）T细胞疗法已攻克多种血液恶性肿瘤，实现了长达十年的缓解，但在实体瘤治疗方面仍面临挑战。通过电穿孔或脂质纳米颗粒（LNP）递送编码CAR的mRNA对CAR T细胞进行非病毒工程改造，可实现高效但短暂的CAR表达，这对现有的基于mRNA的CAR T细胞评估临床前模型的充分性提出了挑战。本研究提出了一种独特的三管齐下的方法，该方法将基于mRNA的CAR T细胞、多靶点靶向胶质母细胞瘤（GBM）相关受体以及最大程度的手术切除相结合，作为一种针对实体瘤的基于mRNA的CAR T细胞临床前评估的新型且易于转化的平台。我们对使用不同扩增条件、mRNA递送方法或联合方法产生的基于mRNA的CAR T细胞进行了直接比较和分析。除了强大的细胞毒性外，我们的研究结果还揭示了抗肿瘤疗效的治疗窗口，以及在接受最大程度手术切除并局部注射多价CAR T细胞（MVCAR）的GBM异种移植小鼠模型中实现强大且持久的完全缓解。在5天扩增的T细胞与静止T细胞中，这种疗效明显更好。有趣的是，在GBM原位切除模型中，MVCAR T细胞优于表达相同CAR scFv组合的汇集CAR T细胞（CARPool）。
