Abstract
Patients with recurrent high-grade glioblastoma have a median survival of 6-8 months, with limited therapeutic options. In recent years, interest has grown in applying chimeric antigen receptor T (CAR-T) cells to solid cancers, including advanced gliomas. Here we generated off-the-shelf CRISPR-Cas9–edited IL-13Rα2-specific allogeneic universal CAR-T cells (MT026) by disrupting the endogenous TCR to prevent graft-versus-host disease and knocking out HLA class I molecules to mitigate the host-versus-graft response, and observed minimal NK-cell–mediated rejection in preclinical studies. In a first-in-human, single-center, open-label investigator-initiated trial (ChiCTR2000028801) in patients with high-grade glioma with prior therapy failure and short life expectancy, intrathecal injection of MT026 via lumbar puncture (1.0-3.0×10^7 cells per dose) demonstrated favorable tolerability and safety (primary outcome), pharmacokinetic characteristics, and preliminary clinical activity (secondary outcomes). Among the five patients enrolled, one achieved a complete response and three achieved partial responses. No grade ≥3 adverse events were observed; the predominant treatment-related toxicities were grade 1-2 pyrexia, hypoxia, and vomiting. Trial enrolment was halted after enrolment of the first five patients, however these preliminary clinical data support the potential benefit of locally administered allogeneic universal CAR-T cell therapy for recurrent glioblastoma.
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Data availability
The off-target detection datasets—comprising GUIDE-seq, PEM-seq, AID-seq, and amplicon sequencing—have been deposited in the Genome Sequence Archive in the National Genomics Data Center, China National Center for Bioinformation, under accession code HRA015321. To protect donor privacy concerning HLA genetic sequences, these data are under controlled access for two years following publication. During this two-year controlled-access period, qualified researchers may request access through the GSA controlled-access system or by contacting the corresponding author, Yulun Huang (Y.H.), for legitimate scientific purposes. Additionally, since the sequencing primers used in the data analysis methods implicate donor HLA privacy, all inquiries concerning data analysis methods must be addressed directly to the corresponding author. A copy of the study protocol is available in the Supplementary Information file. Individual de-identified participant data, including clinical information, imaging data, and treatment responses, are provided in the manuscript and/or the Supplementary Information. Additional de-identified participant-level data may be made available for academic, non-commercial research purposes upon reasonable request to the corresponding author Yulun Huang (Y.H.), subject to institutional approvals and a data-sharing agreement. All other data supporting the findings of this study are available in the article, its supplementary files and source data and/or from the corresponding author upon reasonable request. Source data are provided with this paper.
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Acknowledgements
This work was supported by foundation of Chongqing International Institute for Immunology (2021YZH01 to X-Y. S), National Natural Science Foundation of China (No. 82173279, 82472981 to Y-L.H), and National Science and Technology Resource Sharing Service Platform (YCZYPT[2020]06-1 to Y-L.H), Suzhou Medical and Health Innovation Project (CXYJ2024A05 to Y-L.H), Suzhou Industrial Park Healthcare Talent Support Initiative (2024-54 to Y-L.H), Gusu Talent Program(2024-105 to Y-L.H), and the National Natural Science Foundation of China (82350112 to H-B.L.). These funders provided support for study design, data collection and analysis, and manuscript preparation. We thank the patients who participated in this trial and their families for making this trial possible; the study team, caregivers, and other personnel for their individual professional assistance; and the staff of the department of technology, T-MAXIMUM Pharmaceuticals (Suzhou) Co., Ltd., for technical support. We also extend our gratitude to Yijin Li and Xiu Zhao from T-MAXIMUM Pharmaceuticals for their valuable insights and fruitful discussions that significantly contributed to the development of this study.
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Y.H., X.S., and Y.W. conceived the clinical trial concept. ZhongW., X.L, ZhiminW., J. L., X. Y., H.Z. and Z.X. enrolled patients in the clinical trial, X.Z., X.R., Y.L., X.L., J.S., Z.B., and L.H. evaluated toxicity and participated in critical discussions, as well as manuscript writing and editing. W.G. performed the pathology assessments. H.L. and L.W. performed the immunological assays. X.J., YangZ., and J.C. conducted formal analysis, data visualization, writing—original draft and writing—review and editing. X.S., YuZ., J.G., X.M., and Y.Wang obtained resources, managed data, performed project administration, and performed writing—review and editing. Y.W. and Y.H. contributed to writing—original draft, writing—review and editing, funding acquisition, formal analysis, and data curation.
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Li, X., Shang, X., Liu, J. et al. Intrathecal CRISPR-edited allogeneic IL-13Rα2 CAR T Cells for recurrent high-grade Glioma: preclinical characterization and phase I trial. Nat Commun (2026). https://doi.org/10.1038/s41467-025-68112-6
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DOI: https://doi.org/10.1038/s41467-025-68112-6
