Crp3 modulates aorta biomechanical properties and smooth muscle cells mechanotransduction during aging process.

• Grant number: 24/12223-9
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: January 01, 2025
• End date: December 31, 2027
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: Ayumi Aurea Miyakawa Yamaguchi
• Grantee: Kawane Fabricio Moura
• Host Institution: Instituto do Coração Professor Euryclides de Jesus Zerbini (INCOR). Hospital das Clínicas da Faculdade de Medicina da USP (HCFMUSP). Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated research grant: 23/03079-9 - Crp3 is a key element for vascular smooth muscle cells mechanotransduction and functional cell phenotype, AP.R

Abstract

Focal adhesions are structures capable of sensing changes in the extracellular matrix (ECM) and modulating the tension of actin fibers, thereby altering cellular phenotype. The factors influencing this process are not fully understood, and in our Fapesp project 2023/03079-9, we are testing the hypothesis that Crp3 modulates signaling in focal adhesions, influencing the contractility of aortic smooth muscle cells (SMCs) and cellular and ECM mechanical properties. During aging, there is fragmentation of elastic lamellae, increased collagen deposition, leading to vascular wall stiffening. It is known that smooth muscle cells (SMCs) in aging exhibit larger focal adhesions and increased actin fiber quantity. However, this does not reflect in increased contractile response or increased signaling from focal adhesions. Preliminary data demonstrate that the absence of Crp3 in SMCs mimics the behavior of aged cells, with larger focal adhesions and without modification of their signaling. To better understand the role of Crp3 in modulating SMC phenotype and consequently vascular phenotype, SMCs and aortas from young rats will be evaluated and compared with wild-type and Crp3-KO aged animals. Focal adhesion-mediated signaling will be evaluated in SMCs extracted from young and aged animals. Cellular stiffness and adhesion strength will be assessed by atomic force microscopy (AFM), and contractile properties will be evaluated by collagen gel contraction assay. Functional study of the aorta will be performed through vascular reactivity using vasoactive drugs and using a pressure myograph to evaluate aortic resistance and stiffness in young and aged animals compared to wild-type and Crp3-KO animals. Additionally, the expression of extracellular matrix components such as collagen and elastin will be evaluated. Cellular information on SMC phenotype will be associated with functional and structural responses of the aorta to verify the contribution of cellular findings in the tissue context. Thus, we will evaluate the participation of Crp3 in aortic biomechanics and clarify the underlying mechanisms of vascular diseases associated with mechanical stress, enabling the development of new therapeutic approaches for vascular stiffening observed especially during aging.