Generation of metabolically active NK cells, knockout for the Cish gene, and establishment of non-viral gene transfer strategies for the generation of CAR NK cells (Off-the-shelf).

• Grant number: 22/16140-5
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: December 01, 2023
• End date: November 30, 2025
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Virginia Picanço e Castro
• Grantee: Mara Elisama da Silva Januário
• Host Institution: Hemocentro de Ribeirão Preto. Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da USP (HCMRP). Secretaria da Saúde (São Paulo - Estado). Ribeirão Preto , SP, Brazil
• Associated research grant: 20/07055-9 - Nucleus of Cellular Therapy - NuTeC, AP.NPOP

Abstract

Despite being successful, we need to overcome the limitations of CAR therapy with T cells so that this immunotherapy can treat more patients with reduced cost and increased biosafety. The implementation of this therapy with NK cells brings the possibility of an "Off the shelf" allogeneic therapy, ready and available for treating patients. In addition to NK cells intrinsically eliminating tumor cells, there is no risk of triggering the graft-versus-host disease because, unlike T cells, NK cells can exert their function and kill tumor cells without restriction to MHC molecules. It is known that the cytokine IL-15 is essential in promoting the differentiation and maturation of NK cells. Preliminary results from our research group demonstrated in vivo the enhanced potential of CAR NK cells expressing IL-15, and it is ± receptor. Given this, a possibility that can potentiate the cytotoxic response of CAR NK cells is the generation of NK cell knockout for the Cish gene. The absence of the CIS protein, responsible for inhibiting IL-15 signaling to NK cells, will confer hypersensitivity to IL-15 in NK cells, allowing them to have more remarkable survival, proliferation, and effector functions. We hypothesize that using non-viral vectors will generate safer CAR NK cells for clinical use at a lower cost and with greater therapeutic efficacy. Furthermore, the Cish gene knockout NK cells we will generate expressing CAR can further enhance the efficacy and in vivo persistence of allogeneic NK therapies. Thus, this research project proposes generating CAR NK cells with more potent cytotoxic action, safer and lower cost, aiming to cover more patients for treatment.