Identification of genetic modifiers enhancing B7-H3-targeting CAR T cell therapy against glioblastoma through large-scale CRISPRi screening

Li，Xing1,2; Sun，Shiyu1,4; Zhang，Wansong1,2; Liang，Ziwei1; Fang，Yitong1,2; Sun，Tianhu1,2; Wan，Yong3; Ma，Xingcong4; Zhang，Shuqun4; Xu，Yang1; Tian，Ruilin1,2

2024-12-01

10.1186/s13046-024-03027-6

Journal of Experimental and Clinical Cancer Research   影响因子和分区

1756-9966

Background: Glioblastoma multiforme (GBM) is a highly aggressive brain tumor with a poor prognosis. Current treatment options are limited and often ineffective. CAR T cell therapy has shown success in treating hematologic malignancies, and there is growing interest in its potential application in solid tumors, including GBM. However, current CAR T therapy lacks clinical efficacy against GBM due to tumor-related resistance mechanisms and CAR T cell deficiencies. Therefore, there is a need to improve CAR T cell therapy efficacy in GBM. Methods: We conducted large-scale CRISPR interference (CRISPRi) screens in GBM cell line U87 MG cells co-cultured with B7-H3 targeting CAR T cells to identify genetic modifiers that can enhance CAR T cell-mediated tumor killing. Flow cytometry-based tumor killing assay and CAR T cell activation assay were performed to validate screening hits. Bioinformatic analyses on bulk and single-cell RNA sequencing data and the TCGA database were employed to elucidate the mechanism underlying enhanced CAR T efficacy upon knocking down the selected screening hits in U87 MG cells. Results: We established B7-H3 as a targetable antigen for CAR T therapy in GBM. Through large-scale CRISPRi screening, we discovered genetic modifiers in GBM cells, including ARPC4, PI4KA, ATP6V1A, UBA1, and NDUFV1, that regulated the efficacy of CAR T cell-mediated tumor killing. Furthermore, we discovered that TNFSF15 was upregulated in both ARPC4 and NDUFV1 knockdown GBM cells and revealed an immunostimulatory role of TNFSF15 in modulating tumor-CAR T interaction to enhance CAR T cell efficacy. Conclusions: Our study highlights the power of CRISPR-based genetic screening in investigating tumor-CAR T interaction and identifies potential druggable targets in tumor cells that confer resistance to CAR T cell killing. Furthermore, we devised targeted strategies that synergize with CAR T therapy against GBM. These findings shed light on the development of novel combinatorial strategies for effective immunotherapy of GBM and other solid tumors.

B7-H3 CAR T cell CRISPR screening Glioblastoma multiforme TNFSF15

SCI

英语

第一 ; 通讯

CLINICAL MEDICINE

2-s2.0-85189152664

Scopus

1.School of Medicine,Southern University of Science and Technology,Shenzhen,Guangdong Province,518055,China
2.Key University Laboratory of Metabolism and Health of Guangdong,Southern University of Science and Technology,Shenzhen,Guangdong Province,518055,China
3.Department of Neurosurgery,Shenzhen People’s Hospital,Shenzhen,Guangdong,518020,China
4.Department of Oncology,The Second Affiliated Hospital of Xi’an Jiaotong University,Xi’an,Shaanxi Province,710004,China

Li，Xing,Sun，Shiyu,Zhang，Wansong,et al. Identification of genetic modifiers enhancing B7-H3-targeting CAR T cell therapy against glioblastoma through large-scale CRISPRi screening[J]. Journal of Experimental and Clinical Cancer Research,2024,43(1).

Li，Xing.,Sun，Shiyu.,Zhang，Wansong.,Liang，Ziwei.,Fang，Yitong.,...&Tian，Ruilin.(2024).Identification of genetic modifiers enhancing B7-H3-targeting CAR T cell therapy against glioblastoma through large-scale CRISPRi screening.Journal of Experimental and Clinical Cancer Research,43(1).

Li，Xing,et al."Identification of genetic modifiers enhancing B7-H3-targeting CAR T cell therapy against glioblastoma through large-scale CRISPRi screening".Journal of Experimental and Clinical Cancer Research 43.1(2024).