Saha Tushara, Saha Rudra Prasad, Singh Manoj Kumar, Priya Kanu, Singh Shareen, Rajeev Mithul, Bhattacharya Debasmita, Nag Moupriya, Lahiri Dibyajit
Department of Biotechnology, School of Life Science & Biotechnology, Adamas University, Kolkata, 700126, India.
Centre for Phytochemical Research, Department of Life Science, School of Basic Science and Research, Sharda University, Greater Noida, UP, India.
Mol Biol Rep. 2025 Jun 14;52(1):596. doi: 10.1007/s11033-025-10674-1.
Chimeric antigen receptor T (CAR-T) cell therapy has become a milestone in the management of B cell lineage acute lymphoblastic leukemia. Yet, the traditional method-dependent on ex vivo manipulation, amplification, and reinfusion of autologous T cells-is high-cost, low-scalability, and severely immune-related toxicity. Here, we report a new nano-immunoengineering platform that allows in vivo production of chimeric antigen receptor T cells through the use of functionalized nanoparticles carrying clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) gene editing elements. These nanoparticles are engineered to specifically target blood circulating T lymphocytes and deliver CRISPR/Cas9 complexes that have the ability to integrate chimeric antigen receptor constructs into the TRAC locus and knock out immune checkpoint genes like programmed cell death protein 1 (PD-1) simultaneously. Targeted delivery, endosomal escape, and efficient genome editing with minimal off-target effects are ensured through gold-based and DNA nanostructure-based carriers. Preclinical models show effective in vivo programming of functional chimeric antigen receptor T cells with vigorous antitumor efficacy, improved persistence, and decreased cytokine release syndrome. This method is a revolutionary breakthrough in cancer immunotherapy that provides a scalable, economical, and clinically flexible replacement for conventional chimeric antigen receptor T cell production.
嵌合抗原受体T（CAR-T）细胞疗法已成为B细胞系急性淋巴细胞白血病治疗的一个里程碑。然而，传统方法依赖于体外对自体T细胞的操作、扩增和回输，成本高、可扩展性低，且存在严重的免疫相关毒性。在此，我们报告了一种新的纳米免疫工程平台，该平台通过使用携带成簇规律间隔短回文重复序列（CRISPR）和CRISPR相关蛋白9（Cas9）基因编辑元件的功能化纳米颗粒，实现体内嵌合抗原受体T细胞的产生。这些纳米颗粒经过工程改造，能够特异性靶向血液循环中的T淋巴细胞，并递送CRISPR/Cas9复合物，该复合物能够将嵌合抗原受体构建体整合到TRAC基因座中，并同时敲除程序性细胞死亡蛋白1（PD-1）等免疫检查点基因。通过基于金和基于DNA纳米结构的载体，确保了靶向递送、内体逃逸以及具有最小脱靶效应的高效基因组编辑。临床前模型显示，功能性嵌合抗原受体T细胞在体内的编程有效，具有强大的抗肿瘤功效、更好的持久性以及降低的细胞因子释放综合征。该方法是癌症免疫疗法的一项革命性突破，为传统嵌合抗原受体T细胞生产提供了一种可扩展、经济且临床灵活的替代方案。
