Integrated mechanisms of protein turnover to prevent proteostasis decline with the aging process

• Grant number: 23/02988-5
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: November 01, 2023
• End date: October 31, 2024
• Field of knowledge: Biological Sciences - Physiology - General Physiology
• Principal Investigator: Adelino Sanchez Ramos da Silva
• Grantee: Ivo Vieira de Sousa Neto
• Supervisor: Benjamin Miller
• Host Institution: Escola de Educação Física e Esporte de Ribeirão Preto (EEFERP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Institution abroad: Oklahoma Medical Research Foundation (OMRF), United States
• Associated to the scholarship: 21/14233-3 - Effects of PGC-1alpha conditional deletion and exercise on mitochondrial biogenesis, autophagy and rev-erb-alpha pathways in mouse skeletal muscle: a proteomics approach, BP.PD

Abstract

Proteostatic quality control mechanisms fail with advancing age, resulting in the accumulation of damaged and dysfunctional proteins. However, our understanding is limited because the prior investigations treated tissues as a homogenous collection of cells. Therefore, a critical unknown is how individual cell types within a tissue influence overall tissue proteostasis. The overall goal of this study will be to elucidate previously unexplored mechanisms of protein turnover for proteostatic maintenance. We will accomplish this goal by examining cell-type specific changes in individual protein turnover with aging or treatments that slow aging through direct inhibition of mTOR (rapamycin) or independent of direct mTOR inhibition (17±-estradiol (17±-E2)). For this, the following specific goals will be used: 1) Determine cell-type specific proteins susceptible to proteostatic decline with aging. 2) Determine how a treatment that inhibits mTOR improves proteostasis. 3) Determine how a treatment that does not directly inhibit mTOR improves proteostasis. We will use novel mouse models (NuTRAP-Cre; i.e HSA-myocyte, Pax7-satellite cell, Neuron-CamkIIa, and astrocyte-Aldh1l1) to isolate DNA, protein and mitochondria in a cell-type-specific manner. Our new approaches use the combination of deuterium oxide labeling and discovery or targeted proteomics. The methodologies used in this project will be single-cell, intact nuclear isolation, mass spectrometry and protein synthesis rate. Our study can contribute to a better understanding of the basic biology of aging and clarify more profoundly the molecular networks behind physiological adaptations promoted by protein turnover. Therefore, the findings open new perspectives and insights for studies based on complex mechanisms that govern cellular longevity. (AU)