Preprint / Version 1

AAV variant enables human T cell engineering in vivo

This article is a preprint and has not been certified by peer review.

Authors

    Zhike Lu,  
    Zhike Lu
    Ke Ni,  
    Ke Ni
    • Westlake Genetech
    Wenjun Liu,   Qingkai Song,   Rong Zheng,   Ming Wei,   Yinling Zhang,   Jing Wang,   Lina Wei,   Chenlu Wu,   Qingfeng Zhang,   Jiamei Wu,   Ruijie Zhu,   Shuai Ding,   Chunyu Cheng,   Yanyi Cong,   Yinxia Xu,   Baorui Kong,   Shanshan Wu,   Gang Wang,   Xiaojuan Wang,   Yalin Wang,   Xu Qian,   Ruixia Deng,   Hui Chen,   Yan Li,  
    Yan Li
    Lijia Ma
    Lijia Ma
Categories
Bioengineering & Biotechnology
Keywords
AAV capsid engineering; In vivo CAR-T

Abstract

Autologous chimeric antigen receptor T (CAR-T) cell therapy has demonstrated therapeutic effectiveness in hematologic malignancies and autoimmune diseases. However, the manufacturing complexity and the required lymphodepletion hindered its wide clinical application. Engineering human T cells in vivo holds promise to conquer these limitations but requires effective T cell-targeted CAR delivery with demonstrated safety. Here, we show that an engineered AAV6 variant, AAV6-M2, can enable in vivo CAR expression in human T cells following systemic administration in a Humanized Immune System (HIS) mouse model. AAV6-M2-CD19CAR turned up to 85.2% of human CD8+ T cells into CAR-T cells across multiple organs six weeks post-AAV injection. In HIS mice exhibiting systemic lupus erythematosus (SLE)-like symptoms, AAV6-M2-CD19CAR treatment effectively depleted B cells in both peripheral blood and tissues, accompanied by improved lupus pathologies. Importantly, systemic delivery of AAV6-M2 resulted in significant liver de-targeting, with viral genome levels in the liver reduced by over two orders of magnitude in both mice and cynomolgus macaque compared to the wild-type AAV. Through CRISPR screening, cryo-EM structural analysis, and molecular docking, we identified CD62L as a key mediator of AAV6-M2's enhanced transduction to human T cells, enabling CAR delivery without the need for prior T cell activation. These findings established that AAV-mediated CAR delivery can generate functional human CAR-T cells in vivo, with mechanistic insights into the selective targeting of T cells. This work highlights engineered AAV vectors as a promising platform for in vivo CAR-T therapy and expands the therapeutic landscape of AAV beyond inherited diseases.

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DOI:

https://doi.org/10.65215/LTSpreprints.2026.01.27.000102

Submission ID:

102

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Posted

2026-01-27

How to Cite

Lu, Z., Ni, K., Liu, W., Song, Q., Zheng, R., Wei, M., Zhang, Y., Wang, J., Wei, L., Wu, C., Zhang, Q., Wu, J., Zhu, R., Ding, S., Cheng, C., Cong, Y., Xu, Y., Kong, B., Wu, S., … Ma, L. (2026). AAV variant enables human T cell engineering in vivo. LangTaoSha Preprint Server. https://doi.org/10.65215/LTSpreprints.2026.01.27.000102

Declaration of Competing Interests

Details of all competing interests to be disclosed are as follows:

Westlake Genetech and Westlake University share intellectual property based on the AAV variants of this study. Z.L. and L.M. are co-founders of Westlake Genetech. K.N. was a postdoc in Lijia Ma’s Lab at Westlake University when this project started. K.N.. Y.L. is currently consulting for GemPharmatech Co.

Copyright

The copyright holder for this preprint is the author/funder.

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.