Foster Jessica B, Madsen Peter J, Harvey Kyra, Griffin Crystal, Stern Allison, Patterson Luke, Joshi Nikhil, Dickson Conor, McManus Olivia, Beaubien Ezra, Wilson Cullen, Beale David R, Baubet Valerie, Goel P Payush N, Vitanza Nicholas A, Nazarian Javad, Koptyra Mateusz, Storm Phillip B, Resnick Adam C
Division of Oncology, Children's Hospital of Philadelphia, Philadelphia, PA.
Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA.
Neuro Oncol. 2025 May 24. doi: 10.1093/neuonc/noaf115.
Diffuse midline glioma (DMG) and high grade glioma are devastating pediatric central nervous system tumors that remain incurable. Recent chimeric antigen receptor (CAR) T cell studies have shown proof of concept and early signs of efficacy against DMG targeting GD2. Prior work and ongoing clinical trials have focused on using viral vectors to create permanent CAR T cells. However, virally transduced GD2-directed CAR T cells have shown significant neurotoxicity in both pre-clinical models and human trials.
We evaluated transient CAR T cells targeting GD2 created with mRNA, assessing for efficacy and safety in cell line, organoid, and in vivo xenograft models with repetitive intratumoral dosing.
We show that mRNA GD2-directed CAR T cells are active against both cell lines and organoid models of DMG and high grade glioma in vitro. Cytotoxicity consistently abates over 9 days, highlighting the potential to avoid toxicity from persistent T cell activity. In both pontine and thalamic DMG xenograft models, repeated doses of mRNA GD2-directed CAR T cells were titrated down to maintain therapeutic effect without causing neurologic toxicity.
Our results demonstrate the utility of transient mRNA CAR T cells delivered intratumorally to provide effective tumor killing with a defined half-life, allowing for modulation of the dose and potential side effects. We anticipate this study will expand the use of CAR T cell therapy for DMG and other central nervous system tumors and non-malignant disorders, where concern for toxicity from permanently expressing CAR T cells may hinder development.
弥漫性中线胶质瘤（DMG）和高级别胶质瘤是毁灭性的儿科中枢神经系统肿瘤，仍然无法治愈。最近的嵌合抗原受体（CAR）T细胞研究已经证明了针对靶向GD2的DMG的概念验证和疗效的早期迹象。先前的工作和正在进行的临床试验都集中在使用病毒载体来创建永久性CAR T细胞。然而，病毒转导的靶向GD2的CAR T细胞在临床前模型和人体试验中均显示出显著的神经毒性。
我们评估了用mRNA创建的靶向GD2的瞬时CAR T细胞，通过重复瘤内给药在细胞系、类器官和体内异种移植模型中评估其疗效和安全性。
我们表明，mRNA靶向GD2的CAR T细胞在体外对DMG和高级别胶质瘤的细胞系和类器官模型均有活性。细胞毒性在9天内持续减弱，突出了避免持续性T细胞活性导致毒性的潜力。在桥脑和丘脑DMG异种移植模型中，重复剂量的mRNA靶向GD2的CAR T细胞被滴定降低以维持治疗效果而不引起神经毒性。
我们的结果证明了瘤内递送瞬时mRNA CAR T细胞的效用，以提供具有确定半衰期的有效肿瘤杀伤，从而允许调节剂量和潜在的副作用。我们预计这项研究将扩大CAR T细胞疗法在DMG和其他中枢神经系统肿瘤以及非恶性疾病中的应用，在这些疾病中，对永久表达CAR T细胞毒性的担忧可能会阻碍其发展。
