Osteogenic markers and epigenetic reprogramming during the phenotypic transition of the vascular smooth muscle: from contraction to calcification

• Grant number: 19/22255-7
• Support Opportunities: Scholarships in Brazil - Doctorate
• Effective date (Start): December 01, 2019
• Effective date (End): July 31, 2024
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Willian Fernando Zambuzzi
• Grantee: Geórgia da Silva Feltran
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
• Associated research grant: 14/22689-3 - Microvesicle/proteins-mediated paracrine signaling among bone and endothelial cells during bone development and regeneration, AP.JP
• Associated scholarship(s): 22/06879-3 - Impact of the macrophages on the crosstalk with VSMCs in maintaining vascular calcification microenvironment: understanding the role of SIRT & cytokines, BE.EP.DR

Abstract

The vascular system is responsible for transporting fluids and is properly responsible for moving large volumes of blood throughout the body. All blood vessels have the same basic anatomical structure, composed of a single concentric layer of endothelial cells, surrounded by a secondary layer composed of smooth muscle cells. It is a dynamic tissue with exceptional physiological activities, but it is involved with the installation of vascular pathologies, such as vascular calcification, which is characterized by an ectopic deposition of inorganic calcium phosphate crystals in the arterial tissue leading to an increase risk of morbidity and mortality. It is a pathology of multifactorial etiology, but it has been widely associated with chronic kidney deficiency, which correlates with disturbances in calcium and phosphate metabolism, environmental factors that trigger ectopic calcification. Some authors have already pointed out that vascular calcification recapitulates morphogenic events of osteogenesis, but its molecular study still needs further investigation, especially bringing epigenetic understanding of this environmental influence on the expression of genes involved in smooth muscle cell calcification. In this sense, the proposition of alternative in vitro models guarantees an easily controlled experimental condition to study the etiology of vascular calcification, and this seems to fill a gap between animal models and human impairment. In this context, the main objective of this proposal will be to investigate if the global epigenetic mechanism is involved in the activation of osteogenic marker genes in smooth muscle cells during the calcification process. After standardization of vascular calcification models in 2D and 3D culture, we will perform gene expression analysis of osteogenic markers, analysis of global epigenetic mechanisms through methylation and hydroxymethylation, western blotting for classic markers of these events, and validate the findings in the osteogenic markers, cell culture, in an animal model already described in the literature for the installation of Chronic Kidney Disease. It is noteworthy that this request is financially covered by the JP project, process 2014/22689-3. We have also planned a BEPE period as a partner abroad to develop in vivo objectives, which will add technical and experimental values to the development of projects at the Chemistry and Biochemistry Department, IBB-UNESP. (AU)