Exploiting the mammary matrisome: analysis of the extracellular matrix under healthy and endocrine disruption conditions

• Grant number: 24/06057-9
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: September 01, 2024
• End date: August 31, 2025
• Field of knowledge: Biological Sciences - Morphology - Histology
• Principal Investigator: Sebastião Roberto Taboga
• Grantee: Thalles Fernando Rocha Ruiz
• Supervisor: Christiani Andrade Amorim
• Host Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil
• Institution abroad: Universitè Catolique de Louvain (UCL), Belgium
• Associated to the scholarship: 22/00521-0 - Bisphenol A-induced mammary carcinogenesis in senile females: influence of hormone therapies on neoplastic development, BP.DR

Abstract

Mammary gland (MG) biology shows high complexity related to stromal, which requires an intense remodeling of extracellular matrix (ECM) components influenced by ovarian-derived hormones. Parity and hormonal profile are intrinsic changes that affect ECM composition. The stromal microenvironment's role could extend to breast cancer development, where alterations in proteoglycans, glycosaminoglycans (GAGs), and types of collagen influence tumor progression. This emphasizes the significance of ECM composition in clinical analysis and therapeutics, not only under carcinogenesis but also in a healthy perspective. Thus, we aim (1) to unveil the matrisome of the human breast and (2) to describe ECM remodeling under disrupted MG exposed to hormonal therapies. In the first task, our research will demonstrate the expression of ECM elements of human breast samples (nulliparous and parous) from Hôpital Universitaire de Bruxelles in cooperation with UCLouvain, Belgium. We will explore collagens (I, III, IV, and V), fibronectin, laminin-111, and -332 expression and localization by Western blotting and microscopy techniques. Atomic Force Microscopy (AFM) and Scanning Electron Microscopy (SEM) will be performed to detect the elasticity and fibrillar content of samples. LC-MS/MS and proteomic analysis will support specific markers for ECM structure and their interactions. Also, we will decode proteoglycans and GAG content by glycomic analysis using high-performance liquid chromatography (HPLC). Second task will employ techniques of first task to determine MG matrisome under endocrine disruption and hormonal replacement therapies (HRT). Our previous results demonstrate the onset of carcinogenesis in the experimental model (Mongolian gerbil) and HRT could affect the tumoral progression. Experimental procedures were completed, and gerbil MG samples were collected. Analysis in UCLouvain will integrate proteomic and Western blotting of ECM components with microscopic techniques to detect tumor-associated collagen signatures and remodeling of basement membrane components (collagen IV, nidogen, perlecan). Glycomic, biochemical, and histochemistry will complete the MG matrisome, spotting changes in proteoglycans/GAGs profile. Thus, combined results from proteomic, glycomic, and microscopy will identify features related to normal breast and disrupted microenvironment, associated with parity conditions and hormonal therapies.