IL-2-inducible T cell kinase deficiency sustains chimeric antigen receptor T cell therapy against tumor cells PMID: 39589809

ADC Targets in Oncology

2026-04-10

IL-2-inducible T cell kinase deficiency sustains chimeric antigen receptor T cell therapy against tumor cells

J Clin Invest. 2024 Nov 26;135(4):e178558. doi: 10.1172/JCI178558.

Zheng Fu ¹ ² ³ ⁴ , Zineng Huang ¹ ⁵ ⁶ , Hao Xu ⁷ , Qingbai Liu ⁸ , Jing Li ⁹ , Keqing Song ¹⁰ , Yating Deng ¹ ⁵ ⁶ , Yujia Tao ¹ ⁵ ⁶ , Huifang Zhang ¹ ⁵ ⁶ , Peilong Wang ¹ ⁵ ⁶ , Heng Li ¹ ⁵ ⁶ , Yue Sheng ¹ ⁵ ⁶ , Aijun Zhou ⁸ , Lianbin Han ³ , Yan Fu ³ , Chenzhi Wang ³ , Saurav Kumar Choudhary ¹¹ , Kaixiong Ye ¹¹ ¹² , Gianluca Veggiani ¹³ , Zhihong Li ¹⁴ ¹⁵ , Avery August ¹⁶ , Weishan Huang ¹³ ¹⁶ , Qiang Shan ⁷ , Hongling Peng ¹ ⁵ ⁶ ¹⁵

Affiliations

1. Department of Hematology, The Second Xiangya Hospital, Central South University, Changsha, Hunan, China.
2. Hubei Jiangxia Laboratory, Wuhan, Hubei, China.
3. MegaRobo Technologies Co. Ltd., Suzhou, China.
4. Xinyi Biotech Co. Ltd., Lingang, Shanghai, China.
5. Institute of Hematology, Central South University, Changsha, Hunan, China.
6. Hunan Engineering Research Center of Cell Immunotherapy for Hematopoietic Malignancies, Changsha, Hunan, China.
7. National Key Laboratory of Immunity and Inflammation, Suzhou Institute of Systems Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, Jiangsu, China.
8. Lianshui People's Hospital of Kangda College Affiliated to Nanjing Medical University, Huai'an, Jiangsu Province, China.
9. Key Laboratory of Cluster Science, Ministry of Education of China, School of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing, China.
10. Tianjin Mogenetics Biotech Co. Ltd., Tianjin, China.
11. Institute of Bioinformatics and.
12. Department of Genetics, Franklin College of Arts and Sciences, University of Georgia, Athens, Georgia, USA.
13. Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, Louisiana, USA.
14. Department of Orthopedics, The Second Xiangya Hospital, Central South University, Changsha, China.
15. Hunan Key Laboratory of Tumor Models and Individualized Medicine, Changsha, Hunan, China.
16. Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.

Abstract

Despite the revolutionary achievements of chimeric antigen receptor (CAR) T cell therapy in treating cancers, especially leukemia, several key challenges still limit its therapeutic efficacy. Of particular relevance is the relapse of Cancer in large part as a result of exhaustion and short persistence of CAR-T cells in vivo. IL-2-inducible T cell kinase (Itk) is a critical modulator of the strength of T cell receptor signaling, while its role in CAR signaling is unknown. By electroporation of CRISPR-associated protein 9 (Cas9) ribonucleoprotein (RNP) complex into CAR-T cells, we successfully deleted Itk in CD19-CAR-T cells with high efficiency. Bulk and single-cell RNA Sequencing analyses revealed downregulation of exhaustion and upregulation of memory gene signatures in ITK-deficient CD19-CAR-T cells. Our results further demonstrated a significant reduction of T cell exhaustion and enhancement of T cell memory, with significant improvement of CAR-T cell expansion and persistence both in vitro and in vivo. Moreover, ITK-deficient CD19-CAR-T cells showed better control of tumor relapse. Our work provides a promising strategy of targeting Itk to develop sustainable CAR-T cell products for clinical use.

Keywords

Cancer immunotherapy; Hematology; Immunology; Leukemias; T cells.