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bioRxiv · 10.1101/2025.06.30.660492

Peptide-Specific CAR T cells Recognize WT1 Promiscuously Presented by Diverse HLA Class II Alleles

Abstract

Chimeric antigen receptor (CAR) technology has revolutionized B-cell malignancy treatment by enabling T cells to effectively recognize and target cancer-specific surface antigens. However, CAR T cells show limited efficacy against other blood cancers and solid tumors due to challenges in identifying suitable surface targets. Here, we present a novel approach to CAR development, targeting the intracellular Wilms tumor 1 (WT1) oncoprotein, cross-presented by surface HLA-class II (HLA-II) alleles. WT1-CAR T cells, derived from an antibody raised solely against a WT1 peptide, recognized the WT1330-348 peptide promiscuously presented by 18 out of 20 tested HLA-II alleles, overcoming traditional HLA restrictions. WT1-CAR T cells specifically recognized leukemic cells in a WT1 and HLA-II-dependent manner and mediated an antitumor response in vitro and in vivo. This innovative approach to CAR T cell development transcends traditional HLA restrictions and offers a promising therapeutic option to a wide and genetically diverse patient population. Statement of significanceThis study describes a novel CAR T therapy approach leveraging the distinctive and shared characteristic of HLA-II-peptide binding promiscuity, enabling targeting of the intracellular oncoprotein WT1 presented across diverse HLA-II families. Our study demonstrates a viable framework for designing CAR T therapies that benefit genetically diverse patient populations.

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BibTeXRIS

Wang, C.-H., Zheng, E. Y. F., Ochi, T., Ohashi, Y., Ihara, F., Fukao, S., Boukhaled, G. M., Wang, B. X., Han, D.-H., Wei, X., Burt, B. D., Saso, K., Matsunaga, Y., Ly, D., Kagoya, Y., Butler, M. O., Minden, M., Hirano, N.. 2025-07-04. Peptide-Specific CAR T cells Recognize WT1 Promiscuously Presented by Diverse HLA Class II Alleles. https://doi.org/10.1101/2025.06.30.660492

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