In vivo gene editing of T-cells in lymph nodes for enhanced cancer immunotherapy

• Published in: Nature communications 2024-11, Vol.15 (1), p.10218-15, Article 10218
• Main Authors: Qu, Jin, Wang, Yuan, Xiong, Chuxiao, Wang, Mingxue, He, Xingdao, Jia, Weibin, Li, Cheuk Yin, Zhang, Tianlong, Wang, Zixun, Li, Wei, Kuang, Becki Yi, Shi, Peng
• Format: Article
• Language: English
• Subjects: 13/44; 38/1; 38/39; 42/109; 631/61/2300/1850; 631/67/1059/2325; 639/166/985; 9/10; Animal models; Animals; Apoptosis; Biotechnology; Cancer; Cancer immunotherapy; Cancer therapies; Cell culture; Cell death; Cell Line, Tumor; Cell survival; CRISPR; CRISPR-Cas Systems - genetics; Editing; Effector cells; Electroporation; Electroporation - methods; Female; Gene Editing - methods; Genetic modification; Genome editing; Humanities and Social Sciences; Humans; Hydrogels; Immune checkpoint inhibitors; Immune Checkpoint Inhibitors - pharmacology; Immune Checkpoint Inhibitors - therapeutic use; Immune response (humoral); Immunotherapy; Immunotherapy - methods; In vivo methods and tests; Lymph nodes; Lymph Nodes - immunology; Lymphatic system; Lymphocytes T; Melanoma; Melanoma - genetics; Melanoma - immunology; Melanoma - therapy; Melanoma, Experimental - genetics; Melanoma, Experimental - immunology; Melanoma, Experimental - therapy; Metastases; Mice; Mice, Inbred C57BL; multidisciplinary; PD-1 protein; Programmed Cell Death 1 Receptor - antagonists & inhibitors; Programmed Cell Death 1 Receptor - genetics; Programmed Cell Death 1 Receptor - metabolism; Science; Science (multidisciplinary); Side effects; Solid tumors; T-Lymphocytes - immunology; Transfection; Tumors
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-54292-0

Immune checkpoint blockade (ICB) therapy, while promising for cancer treatment, faces challenges like unexpected side effects and limited objective responses. Here, we develop an in vivo gene-editing strategy for improving ICB cancer therapy in a lastingly effective manner. The approach uses a conductive hydrogel-based electroporation system to enable nucleofection of programmed cell death protein 1 (PD1) targeted CRISPR-Cas9 DNAs into T-cells directly within the lymph nodes, and subsequently produces PD1-deficient T-cells to combat tumor growth, metastasis and recurrence in different melanoma models in mice. Following in vivo gene editing, animals show enhanced cellular and humoral immune responses along with multi-fold increases of effector T-cells infiltration to the solid tumors, preventing tumor recurrence and prolonging their survival. These findings provide a proof-of-concept for direct in vivo T-cell engineering via localized gene-editing for enhanced cancer immunotherapy, and also unlock the possibilities of using this method to treat more complex human diseases. As an alternative to the systemic delivery of immune checkpoint inhibitors, here the authors develop an in vivo gene-edit strategy using a conductive hydrogel-based electroporation system to enable nucleofection of PD1-targeted CRISPR-Cas9 DNAs into lymph node T-cells, resulting in suppression of PD1 expression and promotion of anti-tumor immune responses in preclinical cancer models.