Glucocorticoid-induced cell-derived matrix modulates transforming growth factor \u03b22 signaling in human trabecular meshwork cells

Felix Yemanyi,Vijay Krishna Raghunathan,Janice Vranka

doi:10.1038/s41598-020-72779-w

Felix Yemanyi, Vijay Krishna Raghunathan + Show 1 more

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https://doi.org/10.1038/s41598-020-72779-w

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Abstract

Aberrant remodeling of trabecular meshwork (TM) extracellular matrix (ECM) may induce ocular hypertensive phenotypes in human TM (hTM) cells to cause ocular hypertension, via a yet unknown mechanism. Here, we show that, in the absence of exogenous transforming growth factor-beta2 (TGFβ2), compared with control matrices (VehMs), glucocorticoid-induced cell-derived matrices (GIMs) trigger non-Smad TGFβ2 signaling in hTM cells, correlated with overexpression/activity of structural ECM genes (fibronectin, collagen IV, collagen VI, myocilin), matricellular genes (connective tissue growth factor [CTGF], secreted protein, acidic and rich in cysteine), crosslinking genes/enzymes (lysyl oxidase, lysyl oxidase-like 2–4, tissue transglutaminase-2), and ECM turnover genes/enzymes (matrix metalloproteinases-MMP2,14 and their inhibitors-TIMP2). However, in the presence of exogenous TGFβ2, VehMs and GIMs activate Smad and non-Smad TGFβ2 signaling in hTM cells, associated with overexpression of α-smooth muscle actin (α-SMA), and differential upregulation of aforementioned ECM genes/proteins with new ones emerging (collagen-I, thrombospondin-I, plasminogen activator inhibitor, MMP1, 9, ADAMTS4, TIMP1); with GIM-TGFβ2-induced changes being mostly more pronounced. This suggests dual glaucomatous insults potentiate profibrotic signaling/phenotypes. Lastly, we demonstrate type I TGFβ receptor kinase inhibition abrogates VehM-/GIM- and/or TGFβ2-induced upregulation of α-SMA and CTGF. Collectively, pathological TM microenvironments are sufficient to elicit adverse cellular responses that may be ameliorated by targeting TGFβ2 pathway.

Highlights

Aberrant remodeling of trabecular meshwork (TM) extracellular matrix (ECM) may induce ocular hypertensive phenotypes in human TM cells to cause ocular hypertension, via a yet unknown mechanism
In the absence of exogenous TGFβ2, glucocorticoid-induced cell-derived matrices (GIMs) had no statistically significant effect on the active form of Smad[3] in human TM (hTM) cells compared with vehicle control matrices (VehMs)
In the absence of exogenous TGFβ2, GIMs had no effect on TIMP1 in hTM cells compared with VehMs

Summary

Introduction

Aberrant remodeling of trabecular meshwork (TM) extracellular matrix (ECM) may induce ocular hypertensive phenotypes in human TM (hTM) cells to cause ocular hypertension, via a yet unknown mechanism. In the presence of exogenous TGFβ2, VehMs and GIMs activate Smad and non-Smad TGFβ2 signaling in hTM cells, associated with overexpression of α-smooth muscle actin (α-SMA), and differential upregulation of aforementioned ECM genes/proteins with new ones emerging (collagen-I, thrombospondin-I, plasminogen activator inhibitor, MMP1, 9, ADAMTS4, TIMP1); with GIM-TGFβ2-induced changes being mostly more pronounced. This suggests dual glaucomatous insults potentiate profibrotic signaling/phenotypes. In the absence of exogenous DEX, whether pathologic matrices such as GIMs can intrinsically modulate Smad- and non-Smad-dependent TGFβ signaling pathways in human TM (hTM) cells remains to be demonstrated. A portion of the findings in this study has been reported elsewhere (Yemanyi F, et al IOVS 2019; 60: ARVO E-abstract 5146)[48]

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