Please use this identifier to cite or link to this item:
http://arks.princeton.edu/ark:/88435/dsp01000003417
Title: | FIBRONECTIN MATRIX AS AN INSTIGATOR OF ENDOTHELIAL CELL DYSFUNCTION |
Authors: | Resnikoff, Henry |
Advisors: | Schwarzbauer, Jean E |
Contributors: | Molecular Biology Department |
Subjects: | Molecular biology Cellular biology |
Issue Date: | 2024 |
Publisher: | Princeton, NJ : Princeton University |
Abstract: | The extracellular matrix (ECM) is a 3-dimensional network of proteins, proteoglycans, and glycoproteins organized into complex structures that support tissue specific cell attachment, arrangement, and behavior. Fibronectin (FN) is a foundational ECM protein that is polymerized into fibrils by cells and helps coordinate cell adhesion, migration, and proliferation. FN is essential for cardiovascular development and supports angiogenesis in adults. Despite its noted role in vasculogenesis, the basement membrane, a highly organized ECM that underlies endothelial cells in established vasculature, notably lacks FN. However, in fibrosis, FN accumulates in the subendothelial space, disrupting the normal basement membrane architecture. Utilizing human umbilical vein endothelial cells (HUVECs) cultured on permeable Transwell membranes, we investigated how FN matrix accumulation impacts endothelial cell organization and function. We found the FN matrix assembled in this model is heterogenous and discontinuous, and sites of FN matrix accumulation, driven by exogenous FN supplementation, correlated with regional alterations to cell distribution and cell cycle entry. Time course analysis revealed that FN accumulation preceded other changes, demonstrating a causal link between FN accumulation and monolayer disruption. Functionally, FN accumulation increased the HUVEC monolayer permeability to large solutes and induced increased MDA-MB-231 cell attachment to the monolayer, due to the exposure of subendothelial matrix. These results demonstrate that FN mediates a disruption of endothelial cell barrier function, supporting the hypothesis that FN is causative of pathological vascular dysfunction in fibrosis. We subsequently demonstrate the utility of a FN targeting probe generated from a novel, phage-display derived peptide, for delivering and maintaining fluorescent signal to the FN matrix, thus expanding the toolbox for analyzing and targeting FN matrix. |
URI: | http://arks.princeton.edu/ark:/88435/dsp01000003417 |
Type of Material: | Academic dissertations (Ph.D.) |
Language: | en |
Appears in Collections: | Molecular Biology |
Files in This Item:
File | Description | Size | Format | |
---|---|---|---|---|
Resnikoff_princeton_0181D_15149.pdf | 33.3 MB | Adobe PDF | View/Download |
Items in Dataspace are protected by copyright, with all rights reserved, unless otherwise indicated.