Proteomic analysis of endothelial cells secretome: study about uric acid and Atherosclerosis development

• Grant number: 19/16224-1
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Effective date (Start): June 01, 2020
• Effective date (End): February 29, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Metabolism and Bioenergetics
• Principal Investigator: Flavia Carla Meotti
• Grantee: Bianca Dempsey Pinto
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 18/14898-2 - Investigations of the redox processes in inflammatory response and associated pathologies, AP.JP2
• Associated scholarship(s): 21/13550-5 - Structural and functional characterization of the mammalian chaperone Asna1 (TRC40), BE.EP.DD

Abstract

Atherosclerosis-related cardiovascular disease is the leading cause of death in Brazil and in other developed countries. Atherogenesis is associated with metabolic disorders such as hyperuricemia, followed by an inflammatory process in the vascular tissue. Oxidation of uric acid by inflammatory peroxidases and generation of reactive intermediates is one of the mechanisms potentially involved in Atherogenesis. The vascular endothelium abundantly expresses a heme peroxidase, peroxidasin (PXDN), which is the enzyme responsible for producing hypobromous acid (HOBr). HOBr is important for collagen crosslinking and extracellular matrix structuring. Given the similarity of PXDN with heme peroxidases already described in uric acid oxidation, it is possible that uric acid is also substrate for PXDN. Oxidation of uric acid by PXDN may cause, in addition to decreased synthesis of hypobromous acid, increased production of reactive species derived from uric acid. Preliminary data from the research group showed that endothelial cells (HUVEC) release a peroxidase capable of reacting with uric acid, but this peroxidase has not yet been identified. Thus, the objective of this project is to identify, through proteomics techniques, which proteins are released by HUVEC cells in search of the peroxidasin. This project will also investigate whether the acid is used as a peroxidasin substrate or acts as an enzyme inhibitor. If substrate, uric acid oxidation products by cell secretome and recombinant peroxidasin will be identified in order to understand the mechanism of uric acid oxidation in this environment. Finally, this project proposes the proteomic investigation of the effect of uric acid and urate hydroperoxide on endothelial cells. The execution of this project seeks to contribute to the understanding of the mechanisms related to vascular homeostasis disorder and Atherosclerosis progression, as well as the role of uric acid in cardiovascular diseases. (AU)