In vivo gene editing of T-cells in lymph nodes for enhanced cancer immunotherapy
Jin Qu, Yuan Wang, Chuxiao Xiong, Mingxue Wang, Xingdao He, Weibin Jia, Cheuk Yin Li, Tianlong Zhang, Zixun Wang, Wei Li, Becki Yi Kuang, Peng Shi
Abstract
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.