Use of anti-CD19 CAR-NK cells expressing IL-15/IL-15R± and IL-27 cytokines as an allogeneic treatment for B-cell cancers

• Grant number: 23/09711-9
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: July 01, 2024
• End date: January 31, 2027
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Virginia Picanço e Castro
• Grantee: Alison Felipe Bordini Biggi
• 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

The success of CAR-T cells has created enthusiasm to genetically modify NK cells with CAR to increase their ability to eliminate tumor cells. CAR-NK cells have several advantages over CAR-T cells. First, unlike CAR-T cells, CAR-NK cells retain an intrinsic ability to recognize and target tumor cells through their native receptors, making tumor cell evasion less likely through down-regulation of the CAR target antigen. Second, CAR-NK cells do not undergo in vivo clonal expansion or immunological rejection, therefore, they do not cause the cytokine release syndrome (CRS) seen in many clinical trials of CAR-T. Finally, NK cells have less stringent requirements for HLA matching and do not have the potential to cause graft-versus-host disease (GVHD), an important risk posed by CAR-T cell immunotherapy, which makes it possible for CAR-NK cells to be ready for off-the-shelf therapeutic use. However, NK survival and cell expansion are still challenging obstacles. Our hypothesis is that new fourth-generation CAR constructs, modulating IL-15 or IL-27 pathways, may promote significant increases in NK cell proliferation, activation, cytokine secretion and tumor cytolytic activity. In order to test this hypothesis, our research group developed anti-CD19 CARs co-expressing soluble IL-15 or IL-15 complexed to its RA receptor and IL-27 (fourth-generation CARs). Our data highlighted that these three new vectors (4th generation) were more efficient than CAR.19 (second generation) in NK-92 cells. However, molecular mechanisms related to the activation and response of these constructs are not yet elucidated. Based on our preliminary data, the aim of this study is to evaluate the transcriptome profile of NK-92 modified with the different constructs, as well as to identify signaling pathways and immunological checkpoint molecules that can be modulated by these cytokines. Furthermore, this work aims to evaluate the cytotoxic potential of these vectors in primary NK cells in vitro and in vivo.