Clinical Development of Senza5TM CART5: a Novel Dual Population CD5 CRISPR-Cas9 Knocked out Anti-CD5 Chimeric Antigen Receptor T Cell Product for Relapsed and Refractory CD5+ Nodal T-Cell Lymphomas

作者：Ruchi P. Patel, Guido Ghilardi, Patrizia Porazzi, Steven Yang, Dean Qian, Raymone Pajarillo, Michael Wang, Yunlin Zhang, Stephen J. Schuster, Stefan K. Barta, Aditya Nimmagadda, Adam E. Snook, Nicholas A. Siciliano, Marco Ruella · 发表于：Blood · 年份：2022 · DOI：10.1182/blood-2022-166605 · 被引用次数：8 · 研究领域：CAR-T cell therapy research、Viral Infectious Diseases and Gene Expression in Insects、CRISPR and Genetic Engineering

Introduction: Chimeric antigen receptor T-cell (CART) immunotherapy is FDA-approved for B-cell lymphoblastic leukemia, B-cell non-Hodgkin lymphomas, and multiple myeloma. However, CART immunotherapy for T-cell non-Hodgkin lymphomas (T-NHL) is lagging, with only limited clinical results. T-NHL have a poor prognosis and there is an unmet need for effective and long-lasting treatments. We previously presented preliminary preclinical results of anti-CD5 CAR T cells (CART5) that had CD5 knocked out (KO) (Blood (2020) 136 (Supplement 1): 51-52.). CD5 KO not only prevented CART fratricide but also dramatically enhanced CART function. Critical challenges to developing CART cells for T-NHL are CART fratricide and normal T-cell toxicity. In this study, we addressed these challenges by improving manufacturing and including a dual-population technology to mitigate toxicity. We present the development of Senza5TM CART5 an autologous good-manufacturing practice (GMP)-grade product (Fig.1) that will be tested in a phase I clinical trial for relapsed and refractory CD5+ nodal T-cell lymphomas. Methods and Results: We first identified a single guide RNA (sgRNA) that deletes CD5 in T cells with >90% KO efficiency by flow cytometry and ICE analysis. We then assessed the specificity of the KO by performing iGUIDE sequencing. We confirmed high on-target CD5 cleavage with minimal off-targets (CALCP, C20orf85, INPP4B, XPO7, and SLC10A7). These genes have little to no expression in naïve CD4+ and CD...