The Role of Mitochondrial NAD(P)+ Transhydrogenase (NNT) in Neuroinflammatory Response and Glioblastoma Cell Proliferation in Culture

• Grant number: 24/10257-3
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: August 01, 2024
• End date: March 31, 2025
• Field of knowledge: Biological Sciences - Biochemistry - Metabolism and Bioenergetics
• Principal Investigator: Roger Frigério Castilho
• Grantee: Paulo Francisco Veiga Bizerra
• Host Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 23/00229-0 - Pathophysiological roles of mitochondrial NAD(P)+ transhydrogenase, AP.TEM

Abstract

The mitochondrial NAD(P)+ transhydrogenase (NNT) is a transmembrane protein that catalyzes the transfer of hydride between NAD(H) and NADP(H) coupled with proton transport between the intermembrane space and the mitochondrial matrix. This process can occur in both directions and generate either NADPH or NADH (i.e., forward and reverse reactions). Although both reactions are physiologically relevant, the NADPH generated by NNT activity stands out for providing reducing power to peroxide detoxification systems and participating in anabolic reactions. The current project aims to elucidate the role of NNT in neuroinflammation and the proliferation and viability of astrocytes and glioblastoma cells in culture. To achieve this, comparative studies will be conducted using cultures of microglial cells and astrocytes obtained from C57BL/6 mice with a spontaneous mutation in the Nnt gene (C57Unib.B6-Nnt-/-), resulting in the absence of protein expression, and from congenic control mice expressing this protein (C57Unib.B6-Nnt+/+). Human glioblastoma cell lines will also be used. The specific objectives of the project are as follows: i) To study the inflammatory response of astrocytes and microglial cells obtained from Nnt+/+ and Nnt-/- mice; ii) To characterize the importance of NNT in redox signaling and proliferation of astrocytes in culture, and the effect of respiratory and oxidative phosphorylation inhibitors in this model; iii) To evaluate the effect of NNT silencing on redox signaling, proliferation, and viability of glioblastoma cells and astrocytes in culture; iv) To study the effect of silencing NNT expression in glioblastoma cells treated with the chemotherapy drug temozolomide or those expressing IDH1 mutation.