Preclinical Efficacy and Safety of CD19CAR Cytokine-Induced Killer Cells Transfected with Sleeping Beauty Transposon for the Treatment of Acute Lymphoblastic Leukemia

• Published in: Human gene therapy Vol. 29; no. 5; p. 602
• Main Authors: Magnani, Chiara F, Mezzanotte, Claudia, Cappuzzello, Claudia, Bardini, Michela, Tettamanti, Sarah, Fazio, Grazia, Cooper, Laurence J N, Dastoli, Giuseppe, Cazzaniga, Giovanni, Biondi, Andrea, Biagi, Ettore
• Format: Journal Article
• Language: English
• Published: United States 01.05.2018
• Subjects: Animals; Antigens, CD19 - genetics; Antigens, CD19 - immunology; Antigens, CD19 - therapeutic use; Cytokine-Induced Killer Cells - immunology; Gene Expression Regulation, Neoplastic - genetics; Genetic Vectors - genetics; Genetic Vectors - therapeutic use; Humans; Immunotherapy, Adoptive; Mice; Precursor Cell Lymphoblastic Leukemia-Lymphoma - genetics; Precursor Cell Lymphoblastic Leukemia-Lymphoma - immunology; Precursor Cell Lymphoblastic Leukemia-Lymphoma - pathology; Precursor Cell Lymphoblastic Leukemia-Lymphoma - therapy; Receptors, Antigen, T-Cell - genetics; Receptors, Antigen, T-Cell - immunology; Receptors, Antigen, T-Cell - therapeutic use; T-Lymphocytes - immunology; Transfection; Xenograft Model Antitumor Assays; cytokine-induced killer cells; chimeric antigen receptor; acute lymphoblastic leukemia; Sleeping Beauty transposon
• ISSN: 1557-7422
• DOI: 10.1089/hum.2017.207

Infusion of patient-derived CD19-specific chimeric antigen receptor (CAR) T cells engineered by viral vectors achieved complete remission and durable response in relapsed and refractory (r/r) B-lineage neoplasms. Here, we expand on those findings by providing a preclinical evaluation of allogeneic non-viral cytokine-induced killer (CIK) cells transfected with the Sleeping Beauty (SB) transposon CD19CAR (CARCIK-CD19). Specifically, thanks to a large-scale 18-day manufacturing process, it was possible to achieve stable CD19CAR expression (62.425 ± 6.399%) and efficient T-cell expansion (23.36 ± 3.00-fold). Frozen/thawed CARCIK-CD19 remained fully functional both in vitro and in an established patient-derived xenograft (PDX) of MLL-ENL rearranged acute lymphoblastic leukemia (ALL). CARCIK-CD19 showed a dose-dependent antitumor response and prolonged persistence in a PDX, bearing the feature of a Philadelphia-like ALL with PAX5/AUTS2 translocation, and in a survival model of lymphoma, achieving complete eradication of disseminated tumors. Finally, the infusion of CARCIK-CD19 proved to be safe and well tolerated in a biodistribution and toxicity model. The infused cells persisted in the hematopoietic and post-injection perfused organs until the end of the study and consisted of CD8 , CD56 , and CAR T cells. Overall, these findings provide important implications for non-viral technology and the proof-of-concept that donor-derived CARCIK-CD19 are indeed effective against relapsed ALL, a possibility that will be tested in Phase I/II clinical trials after allogeneic hematopoietic stem-cell transplantation.