Evaluation of amphregulin as a prognostic biomarker of severity and its participation in the pathophysiology of COVID-19

• Grant number: 22/08458-5
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): August 01, 2022
• Effective date (End): June 30, 2023
• Field of knowledge: Biological Sciences - Pharmacology - Biochemical and Molecular Pharmacology
• Principal Investigator: José Carlos Farias Alves Filho
• Grantee: Robertha Mariana Rodrigues Lemes
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 20/05211-3 - Evaluation of Amphiregulin as a prognostic biomarker of severity and its participation in the pathophysiology of COVID-19, AP.R

Abstract

The outbreak of the new SARS-CoV-2 virus infection became a pandemic in 2020, being called COVID-19. Because it is an acute disease, it resolves itself in most cases. However, some patients have massive damage to alveolar cells, resulting from viral replication and exacerbated local inflammation. On the other hand, in a systemic way, the plasma elevation of pro-inflammatory cytokines, such as IL-1², IL-6, and TNF, can cause acute cardiac and renal lesions, or even favor secondary bacterial infections. Justify: By containing the infection and repairing injured tissues, the orchestration of immune system cells becomes essential for the resolution and cure of diseases. Amphregulin plays a central role in regulating the host's mechanisms of tolerance to infections, so that AREG-/- mice show delays in the restoration of lung function in cases of injury. On the other hand, several studies show that the administration of recombinant AREG potentiates the repair of tissues injured by excessive inflammation. Objective: Thus, the present study aims to evaluate the role of amphregulin in tissue repair and development of pulmonary fibrosis resulting from SARS-COV-2 infection. Methods: 1- Viral expansion and viral quantification in Vero-E2 cells by PFU/mL and TCID50; 2- Intranasal infection of AREG-/-, AREGf/fLysMCre mice and respective controls B6 and littermates AREGf/f and LysMCre, 3-body weight, 4-viral load, 5-assessment of AREG levels and inflammatory mediators by Elisa and qPCR-RT or immunofluorescence (AREG), 6-quantitation of tissue injury biomarkers such as AST, ALT, CK-MB, urea; 7- Histopathological study of vital organs; 8- Analysis of PBMC cells from peripheral blood of mice with cellular markers (CD45, Ly6C, Ly6G, CD11b, F4/80, CD4, CD8, FOXP3) by flow cytometry. Timeline: Initially, the project will focus on the standardization of experimental models of SARS-CoV-2 infection (1st quarter), to later investigate the importance of AREG in the pathophysiology of COVID-19, using an experimental SARS-CoV infection model -2 (2nd, 3rd and 4th quarters). (AU)