TY - JOUR
T1 - Glaucomatous cell derived matrices differentially modulate non-glaucomatous trabecular meshwork cellular behavior
AU - Raghunathan, Vijay Krishna
AU - Benoit, Julia
AU - Kasetti, Ramesh
AU - Zode, Gulab
AU - Salemi, Michelle
AU - Phinney, Brett S.
AU - Keller, Kate E.
AU - Staverosky, Julia A.
AU - Murphy, Christopher J.
AU - Acott, Ted
AU - Vranka, Janice
N1 - Funding Information:
This study was supported by Bright Focus National Glaucoma Research Awards (VKR, JAV), startup funding at UHCO (VKR), and National Institute of Health grants EY003279, EY008247, EY025721 (TSA) and P30 EY010572, and by an unrestricted grant to the Casey Eye Institute from Research to Prevent Blindness, New York, NY. The authors would like to thank Dr. Paul Russell for his valuable insight and discussions through the preparation of this manuscript.
Funding Information:
This study was supported by Bright Focus National Glaucoma Research Awards (VKR, JAV), startup funding at UHCO (VKR), and National Institute of Health grants EY003279, EY008247, EY025721 ( TSA ) and P30 EY010572, and by an unrestricted grant to the Casey Eye Institute from Research to Prevent Blindness , New York, NY. The authors would like to thank Dr. Paul Russell for his valuable insight and discussions through the preparation of this manuscript.
Publisher Copyright:
© 2018 Acta Materialia Inc.
PY - 2018/4/15
Y1 - 2018/4/15
N2 - Ocular hypertension is a causal risk-factor to developing glaucoma. This is associated with stiffening of the trabecular meshwork (TM), the primary site of resistance to aqueous-humor-outflow. The mechanisms underlying this stiffening or how pathologic extracellular matrix (ECM) affects cell function are poorly understood. It is recognized that mechanotransduction systems allow cells to sense and translate the intrinsic biophysical properties of ECM into intracellular signals to control gene transcription, protein expression, and cell behavior. Using an anterior segment perfusion model, we document that there are significantly more low flow regions that are much stiffer, and fewer high flow regions that are less stiff in glaucomatous TM (GTM) when compared to non-glaucomatous TMs (NTM). GTM tissue also has fewer cells overall when compared with NTM tissue. In order to study the role of pathologic ECM in glaucoma disease progression, we conducted studies using cell derived matrices (CDM). First, we characterized the mechanics, composition and organization of fibronectin in ECM deposited by GTM and NTM cells treated with glucocorticosteroids. Then, we determined that these GTM-derived ECM are able to induce stiffening of normal NTM cells, and alter their gene/protein expression to resemble that of a glaucomatous phenotype. Further, we demonstrate that GTM-derived ECM causes endoplasmic reticular stress in NTM. They also became resistant to being reorganized by these NTM cells. These phenomena were exacerbated by ECMs obtained from steroid treated glaucoma model groups. Collectively, our data demonstrates that CDMs represent a novel tool for the study of bidirectional interactions between TM cells and their immediate microenvironment. Statement of Significance: Extracellular matrix (ECM) changes are prevalent in a number of diseases. The precise mechanisms by which changes in the ECM contribute to disease progression is unclear, primarily due to absence of appropriate models. Here, using glaucoma as a disease model, we document changes in cell derived matrix (CDM) and tissue mechanics that contribute to the pathology. Subsequently, we determine the effect that ECMs from diseased and healthy individuals have on healthy cell behaviors. Data emanating from this study demonstrate that CDMs are a potent tool for the study of cell-ECM interactions.
AB - Ocular hypertension is a causal risk-factor to developing glaucoma. This is associated with stiffening of the trabecular meshwork (TM), the primary site of resistance to aqueous-humor-outflow. The mechanisms underlying this stiffening or how pathologic extracellular matrix (ECM) affects cell function are poorly understood. It is recognized that mechanotransduction systems allow cells to sense and translate the intrinsic biophysical properties of ECM into intracellular signals to control gene transcription, protein expression, and cell behavior. Using an anterior segment perfusion model, we document that there are significantly more low flow regions that are much stiffer, and fewer high flow regions that are less stiff in glaucomatous TM (GTM) when compared to non-glaucomatous TMs (NTM). GTM tissue also has fewer cells overall when compared with NTM tissue. In order to study the role of pathologic ECM in glaucoma disease progression, we conducted studies using cell derived matrices (CDM). First, we characterized the mechanics, composition and organization of fibronectin in ECM deposited by GTM and NTM cells treated with glucocorticosteroids. Then, we determined that these GTM-derived ECM are able to induce stiffening of normal NTM cells, and alter their gene/protein expression to resemble that of a glaucomatous phenotype. Further, we demonstrate that GTM-derived ECM causes endoplasmic reticular stress in NTM. They also became resistant to being reorganized by these NTM cells. These phenomena were exacerbated by ECMs obtained from steroid treated glaucoma model groups. Collectively, our data demonstrates that CDMs represent a novel tool for the study of bidirectional interactions between TM cells and their immediate microenvironment. Statement of Significance: Extracellular matrix (ECM) changes are prevalent in a number of diseases. The precise mechanisms by which changes in the ECM contribute to disease progression is unclear, primarily due to absence of appropriate models. Here, using glaucoma as a disease model, we document changes in cell derived matrix (CDM) and tissue mechanics that contribute to the pathology. Subsequently, we determine the effect that ECMs from diseased and healthy individuals have on healthy cell behaviors. Data emanating from this study demonstrate that CDMs are a potent tool for the study of cell-ECM interactions.
KW - Atomic force microscopy
KW - Biomechanics
KW - Cell derived matrices
KW - Extracellular matrix
KW - Glaucoma
KW - Mechanotransduction
KW - Trabecular meshwork
UR - http://www.scopus.com/inward/record.url?scp=85044003474&partnerID=8YFLogxK
U2 - 10.1016/j.actbio.2018.02.037
DO - 10.1016/j.actbio.2018.02.037
M3 - Article
C2 - 29524673
AN - SCOPUS:85044003474
SN - 1742-7061
VL - 71
SP - 444
EP - 459
JO - Acta Biomaterialia
JF - Acta Biomaterialia
ER -