哈佛大学博德研究所Marcela V. Maus研究团队取得一项新突破。他们报道了体内CRISPR筛选鉴定骨髓瘤中CAR-T细胞功能的修饰因子。这一研究成果发表在2025年9月24日出版的国际学术期刊《自然》上。

在这里，该课题组人员在靶向B细胞成熟抗原的CAR - T细胞中进行了体内功能缺失CRISPR筛选，以研究在人类多发性骨髓瘤模型中影响CAR-T细胞持久性和功能的基因。该团队在体外和体内的早期和晚期时间点跟踪了CRISPR文库编辑的T细胞的扩增和持久性，以跟踪基因修饰的CAR-T细胞从制造到在肿瘤中存活的表现。这些筛选揭示了CAR-T细胞扩增和持久性的环境特异性调节因子。在体外，消融RASA2和SOCS1增强了T细胞的扩增，而缺失PTPN2、ZC3H12A和RC3H1则赋予CAR-T细胞在体内的早期生长优势。值得注意的是，该研究团队发现细胞周期蛋白依赖性激酶抑制剂1B（由CDKN1B编码）是一种细胞周期调节剂，是体内晚时间点限制CAR-T细胞适应性的最重要因素。CDKN1B消融增加CAR-T细胞增殖和效应功能，显著提高肿瘤清除率和总生存期。他们的发现揭示了基因干扰随时间和不同环境对CAR-T细胞的不同影响，强调了CDKN1B作为一个有希望的靶标，可以产生高效的多发性骨髓瘤CAR-T细胞，并强调了体内筛选识别基因以增强CAR-T细胞功效的潜力。

据了解，嵌合抗原受体（CAR） T细胞在血液系统恶性肿瘤中非常有效。然而，CAR-T细胞的逐渐丢失会导致许多患者的复发。

附：英文原文

Title: In vivo CRISPR screens identify modifiers of CAR T cell function in myeloma

Author: Knudsen, Nelson H., Escobar, Giulia, Korell, Felix, Kienka, Tamina, Nobrega, Celeste, Anderson, Seth, Cheng, Andrew Y., Zschummel, Maria, Armstrong, Alexander, Bouffard, Amanda, Kann, Michael C., Goncalves, Sadie, Pope, Hans W., Pezeshki, Mitra, Rojas, Alexander, Suermondt, Juliette S. M. T., Phillips, Merle, Berger, Trisha R., Park, Sangwoo, Salas-Benito, Diego, Darnell, Elijah P., Birocchi, Filippo, Leick, Mark B., Larson, Rebecca C., Doench, John G., Sen, Debattama, Yates, Kathleen B., Manguso, Robert T., Maus, Marcela V.

Issue&Volume: 2025-09-24

Abstract: Chimeric antigen receptor (CAR) T cells are highly effective in haematological malignancies1. However, progressive loss of CAR T cells contributes to relapse in many patients2,3,4. Here we performed in vivo loss-of-function CRISPR screens in CAR T cells targeting B cell maturation antigen to investigate genes that influence CAR T cell persistence and function in a human multiple myeloma model. We tracked the expansion and persistence of CRISPR library-edited T cells in vitro and at early and late time points in vivo to track the performance of gene-modified CAR T cells from manufacturing to survival in tumours. The screens revealed context-specific regulators of CAR T cell expansion and persistence. Ablation of RASA2 and SOCS1 enhanced T cell expansion in vitro, whereas loss of PTPN2, ZC3H12A and RC3H1 conferred early growth advantages to CAR T cells in vivo. Notably, we identified cyclin-dependent kinase inhibitor 1B (encoded by CDKN1B), a cell cycle regulator, as the most important factor limiting CAR T cell fitness at late time points in vivo. CDKN1B ablation increased CAR T cell proliferation and effector function, significantly enhancing tumour clearance and overall survival. Our findings reveal differing effects of gene perturbation on CAR T cells over time and in different environments, highlight CDKN1B as a promising target to generate highly effective CAR T cells for multiple myeloma and underscore the potential of in vivo screening for identifying genes to enhance CAR T cell efficacy.

DOI: 10.1038/s41586-025-09489-8

Source: https://www.nature.com/articles/s41586-025-09489-8