Engineering NK-CAR.19 cells with the IL-15/IL-15Rα complex improved proliferation and anti-tumor effect in vivo

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Silvestre, Renata Nacasaki ; Eitler, Jiri ; de Azevedo, Julia Teixeira Cottas ; Tirapelle, Mariane Cariati ; Fantacini, Daianne Maciely Carvalho ; de Souza, Lucas Eduardo Botelho ; Swiech, Kamilla ; Covas, Dimas Tadeu ; Calado, Rodrigo T. ; Montero, Paola Ortiz ; Malmegrim, Kelen Cristina Ribeiro ; Figueiredo, Marxa L. ; Tonn, Torsten ; Picanco-Castro, Virginia

• Document type: Journal article
• Source: FRONTIERS IN IMMUNOLOGY; v. 14, p. 15-pg., 2023-09-25.

Abstract

Introduction: Natural killer 92 (NK-92) cells are an attractive therapeutic approach as alternative chimeric antigen receptor (CAR) carriers, different from T cells, once they can be used in the allogeneic setting. The modest in vivo outcomes observed with NK-92 cells continue to present hurdles in successfully translating NK-92 cell therapies into clinical applications. Adoptive transfer of CAR-NK-92 cells holds out the promise of therapeutic benefit at a lower rate of adverse events due to the absence of GvHD and cytokine release syndrome. However, it has not achieved breakthrough clinical results yet, and further improvement of CAR-NK-92 cells is necessary.Methods: In this study, we conducted a comparative analysis between CD19-targeted CAR (CAR.19) co-expressing IL-15 (CAR.19-IL15) with IL-15/IL-15R alpha (CAR.19-IL15/IL15R alpha) to promote NK cell proliferation, activation, and cytotoxic activity against B-cell leukemia. CAR constructs were cloned into lentiviral vector and transduced into NK-92 cell line. Potency of CAR-NK cells were assessed against CD19-expressing cell lines NALM-6 or Raji in vitro and in vivo in a murine model. Tumor burden was measured by bioluminescence.Results: We demonstrated that a fourth- generation CD19-targeted CAR (CAR.19) co-expressing IL-15 linked to its receptor IL-15/IL-15R alpha (CAR.19-IL-15/IL-15R alpha) significantly enhanced NK-92 cell proliferation, proinflammatory cytokine secretion, and cytotoxic activity against B-cell cancer cell lines in vitro and in a xenograft mouse model.Conclusion: Together with the results of the systematic analysis of the transcriptome of activated NK-92 CAR variants, this supports the notion that IL-15/IL-15R alpha comprising fourth-generation CARs may overcome the limitations of NK-92 cell-based targeted tumor therapies in vivo by providing the necessary growth and activation signals. (AU)

• FAPESP's process: 13/08135-2 - CTC - Center for Cell-Based Therapy
• Grantee: Dimas Tadeu Covas
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 19/25309-0 - Off-the-shelf engineered NK cells for the treatment of Leukemia and Lymphoma treatment
• Grantee: Virginia Picanço e Castro
• Support Opportunities: Regular Research Grants
• FAPESP's process: 20/08279-8 - Establishment of a platform for generating NK-CAR cells with greater therapeutic effectiveness in vitro and in vivo
• Grantee: Júlia Teixeira Cottas de Azevedo
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 20/07055-9 - Nucleus of Cellular Therapy - NuTeC
• Grantee: Rodrigo do Tocantins Calado de Saloma Rodrigues
• Support Opportunities: Research Grants - Problem-Oriented Research Centers in São Paulo