T cells expressing GXMR-CAR and CD19-CAR to control cryptococcosis and B cell neoplasms.

• Grant number: 24/00300-9
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: December 01, 2024
• End date: July 31, 2027
• Field of knowledge: Biological Sciences - Immunology - Applied Immunology
• Principal Investigator: Thiago Aparecido da Silva
• Grantee: Milena Barrocali de Araújo Melo
• Host Institution: Faculdade de Ciências Farmacêuticas (FCFAR). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil

Abstract

Human cryptococcosis, an infection caused by Cryptoccocus spp., was classified by the WHO as a high priority for the development of therapies. Cryptococcosis has an estimated global annual incidence of 1 million cases, being an invasive fungal infection (IFI) with the lowest percentages of investment in research among neglected diseases. The genomic plasticity of Cryptococcus spp. associated with antifungal resistance are factors that compromise the prophylactic strategy with antifungal drugs prior to therapy for hematological malignancies. New therapeutic strategies are necessary to control infections caused by Cryptococcus gattii and Cryptococcus neoformans, that are of greater clinical relevance, considering immunocompromised and immunocompetent individuals. Moreover, patients with lymphoproliferative neoplasms, in treatment or not, are susceptible to infection by Cryptococcus spp.. In the face of success of immunotherapy with CAR (Chimeric Antigen Receptor) technology for the treatment of certain hematological neoplasms and the possibility of using CAR therapy in the context of cryptococcosis, the present proposal aims to modify human T cells to co-express GXMR-CAR (specific for Cryptococcus spp.) and CD19-CAR aiming to control cryptococcosis and the expansion of neoplastic B cells. This approach will begin with the amplification of GXMR-CAR variants through modification of the signal transduction domain, with emphasis on the co-stimulatory molecules CD28, CD137 and iCOS. The GXMR-CAR variants will be expressed in T cells to evaluate the in vitro fungicidal effect against C. neoformans. Subsequently, T cells will be modified to co-express GXMR-CAR and CD19-CAR and the fungicidal effect against C. neoformans and cytotoxic activity against neoplastic B cell lines will be evaluated in vitro. Finally, T cells modified to co-express GXMR-CAR and CD19-CAR will be used in the therapy of NSG mice previously infected with C. neoformans or infused with a neoplastic B cell line. In vivo control of cryptococcosis will be determined by quantifying fungal in the brain and lung, and reduction of neoplastic B cell expansion will be determined by IVIS microscopy. Therefore, the proposal presents an innovative perspective by presenting in an unprecedented way the approach of co-expressing CAR for the treatment of cryptococcosis and B cell neoplasms.