Etude des effets du collagène sur la production fibroblastique de MMPs en vue d'optimiser les modèles 3D endométriaux

Details

Supervisors

Henriet, Patrick

Faculty

Faculté de pharmacie et des sciences biomédicales

Degree label

Master [120] en sciences biomédicales, à finalité approfondie

Abstract

enThe human endometrium is a tissue undergoing complex cyclical remodeling. The initiation of menstruation results from a massive induction of the expression of matrix metalloproteinases (MMPs) responsible for the degradation of the extracellular matrix. Dysregulation of these enzymes can lead to dysfunctional uterine bleeding and the pathogenesis of endometriosis lesions. In order to further investigate these pathologies, three-dimensional (3D) in vitro models are being developed to mimic the endometrium. However, matrices containing type I collagen, the main component of the stroma, degrade in less than six days, limiting their use. It is therefore essential to understand the molecular mechanisms underlying this degradation in order to design stable and physiologically relevant 3D models. My thesis focused on comparing endometrial fibroblasts embedded in type I collagen droplets or Matrigel. First, I characterized cell viability. Second, I continued previous work by the laboratory aimed at exploring the mechanisms responsible for MMP-2 activation in collagen I droplets. The study focused on the subcellular localization of MT1-MMP (the activator of MMP-2) and collagen receptors (integrins α1β1 and α2β1, and DDR-2). Several approaches were combined: gene expression analysis (qPCR), measurement of MMP-2 activation by western blot, fluorescent co-immunolabeling to track intracellular MT1-MMP trafficking, and pharmacological inhibition of the receptors. The results show that the multi-tyrosine kinase inhibitor Dasatinib abolishes MMP-2 activation in collagen, suggesting the involvement of a tyrosine kinase pathway, potentially dependent on DDR-2. Furthermore, analysis of intracellular trafficking reveals that MT1-MMP is retained more in the endoplasmic reticulum (KDEL) in Matrigel droplets, suggesting a reduction in its activating potential at the plasma membrane. These data provide new mechanistic insights into collagen degradation in endometrial 3D models and contribute to the design of more stable and reproducible biomimetic systems.