Analysis of the physiological and pathological role of the klotho protein in a cellular model of Alzheimer's Disease

• Grant number: 21/06759-5
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: April 01, 2022
• End date: April 02, 2026
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: Merari de Fátima Ramires Ferrari
• Grantee: Alan Moreira Henrique
• Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated scholarship(s): 23/13548-6 - CHARACTERIZATION AND THE ROLE OF ANTI-AGING PROTEIN KLOTHO IN HUMAN 3D MONOCYTE-DERIVED MICROGLIA (MDMi) CULTURES, BE.EP.DD

Abstract

Klotho is widely described as a neuroprotective and anti-aging protein and is associated with numerous age-related diseases, including neurodegenerative disorders. Its functions in the kidney are relatively well known. However, despite the beneficial effects it produces in the brain and neurons, its mechanisms of action in the brain tissue remain elusive. In view of this, the present study proposes to evaluate how klotho behaves in cholinergic neurons and its association with physiological and pathological processes in these cells. To this end, cells of the murine N2a line will be differentiated in cholinergic neurons. These cells will be manipulated in terms of KL and APP expression, which may make possible to recognize cellular processes associated to klotho in physiological and pathological contexts. We will analyze the expression and localization of this protein in the cells, the levels of oxidative and endoplasmic reticule stress, how levels of klotho associate with these stresses and with aging in these neurons, as well as mitophagy and autophagic flux and in what way overexpression of APP affects these functions. This project will make use of RT-qPCR, western blot and electron and fluorescence microscopy techniques in order to answer the evoked questions. (AU)