Development of a compact bidirectional promoter-driven dual chimeric antigen receptor (CAR) construct targeting CD19 and CD20 in the Sleeping Beauty (SB) transposon system

• DOI: 10.1136/jitc-2023-008555
• Publication Date: 2024-04-27T14:25:19Z
• Authors: Asmita Khaniya; S M Ali Hossieni Rad; Josh Halpin; Supannikar Tawinwung; Alexander McLellan; Koramit Suppipat; Nattiya Hirankarn
• Keywords: 0303 health sciences; Receptors, Chimeric Antigen; Immune Cell Therapies and Immune Cell Engineering; Antigens, CD19; Neoplasms. Tumors. Oncology. Including cancer and carcinogens; Mice, SCID; Antigens, CD20; Immunotherapy, Adoptive; Xenograft Model Antitumor Assays; Mice; 03 medical and health sciences; Mice, Inbred NOD; Cell Line, Tumor; DNA Transposable Elements; Humans; Animals; Promoter Regions, Genetic; RC254-282

Background A bidirectional promoter-driven chimeric antigen receptor (CAR) cassette provides the simultaneous expression of two CARs, which significantly enhances dual antigen-targeted CAR T-cell therapy. Methods We developed a second-generation directing CD19 and CD20 antigens, incorporating them in head-to-head orientation from promoter using single Sleeping Beauty transposon system. The efficacy T cells was determined vitro against cell lines expressing either, or both, antigens. In vivo antitumor activity tested Raji lymphoma-bearing immunodeficient NOD-scid IL2Rgamma null (NSG) mice. Results Of all promoters, EF-1 α optimally expressed transcripts both sense (CD19-CAR) antisense (GFP.CD20-CAR) directions. Superior cytotoxicity, cytokine production antigen-specific activation were observed CD19/CD20 cells. contrast, unidirectional construct driven by promoter, but self-cleaving peptide-linked showed inferior function. Treatment mice bearing advanced lymphomas with effectively controlled tumor growth extended survival compared group treated targeted Conclusion use promoters vector offers advantages size robust potential to expand other forms gene therapies like