TRPM2 knockout and its impact in an animal model of Parkinson's disease

• Grant number: 22/14846-8
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: September 01, 2023
• End date: August 31, 2024
• Field of knowledge: Biological Sciences - Physiology - General Physiology
• Principal Investigator: Luiz Roberto Giorgetti de Britto
• Grantee: Ana Flávia Fernandes Ferreira
• Supervisor: Hong-Shuo Sun
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: University of Toronto (U of T), Canada
• Associated to the scholarship: 20/02109-3 - Involvement of TRPM2 channel in neurodegeneration and microglial activation in a Parkinson's Disease model induced by 6-hydroxydopamine, BP.DR

Abstract

Parkinson's disease (PD) is a devastating disease clinically diagnosed by motor dysfunctions that affect millions of people worldwide. Among the neuropathological hallmarks are the degeneration of dopaminergic neurons in the substantia nigra pars compacta and the loss of dopamine in its projection area, the striatum. Currently there is no cure for PD, so many researchers aim to develop new therapeutical targets. Transient Receptor Potential Melastatin 2 (TRPM2) is a calcium channel that is expressed in different cell types, including neurons and glial cells. Activated by oxidative stress, the channel appears to be one of the mechanisms involved in cell death. The TRPM2 potential role in neurodegenerative diseases, including Alzheimer's disease and ischemia, has been described. However, more studies are needed to achieve a better understanding of the involvement of TRPM2 in PD. Thus, the aims of this project are (i) to evaluate the effect of TRPM2 inhibition on a PD animal model using genetically modified TRPM2 mouse and (ii) to verify the effect of TRPM2 selective inhibition on microglial and neuronal cells in an in vitro PD model. The in vivo approach involves TRPM2 knockout mice that will be submitted to a stereotaxic surgery to inject 6-hydroxydopamine (6-OHDA) into the right striatum. At days 7, 14, and 21 after surgery the animals will be submitted to motor behavior tests. At day 21, brains will be collected for immunohistochemical assays. For the in vitro approach, a coculture will be established between wild-type neurons and TRPM2 knockout microglia, and vice-versa. The cultures will be treated with 6-OHDA and cell viability, cell death, intracellular calcium, immunohistochemistry, and cytokine levels will all be evaluated. The results will highlight the TRPM2 involvement in PD, which might be important for the establishment of this channel as new pharmacological target for PD. (AU)