Role of ID proteins in BMP4 inhibition of profibrotic effects of TGF-β2 in human TM cells

Avani A. Mody, Robert J. Wordinger, Abbot Clark

Research output: Contribution to journalArticleResearchpeer-review

4 Citations (Scopus)

Abstract

PURPOSE. Increased expression of TGF-β2 in primary open-angle glaucoma (POAG) aqueous humor (AH) and trabecular meshwork (TM) causes deposition of extracellular matrix (ECM) in the TM and elevated IOP. Bone morphogenetic proteins (BMPs) regulate TGF-β2-induced ECM production. The underlying mechanism for BMP4 inhibition of TGF-β2-induced fibrosis remains undetermined. Bone morphogenic protein 4 induces inhibitor of DNA binding proteins (ID1, ID3), which suppress transcription factor activities to regulate gene expression. Our study will determine whether ID1and ID3 proteins are downstream targets of BMP4, which attenuates TGF-β2 induction of ECM proteins in TM cells. METHODS. Primary human TM cells were treated with BMP4, and ID1 and ID3 mRNA, and protein expression was determined by quantitative PCR (Q-PCR) and Western immunoblotting. Intracellular ID1 and ID3 protein localization was studied by immunocytochemistry. Transformed human TM cells (GTM3 cells) were transfected with ID1 or ID3 expression vectors to determine their potential inhibitory effects on TGF-β2-induced fibronectin and plasminogen activator inhibitor-I (PAI-1) protein expression. RESULTS. Basal expression of ID1-3 was detected in primary human TM cells. Bone morphogenic protein 4 significantly induced early expression of ID1 and ID3 mRNA (P < 0.05) and protein in primary TM cells, and a BMP receptor inhibitor blocked this induction. Overexpression of ID1 and ID3 significantly inhibited TGF-β2-induced expression of fibronectin and PAI-1 in TM cells (P < 0.01). CONCLUSIONS. Bone morphogenic protein 4 induced ID1 and ID3 expression suppresses TGFβ2 profibrotic activity in human TM cells. In the future, targeting specific regulators may control the TGF-β2 profibrotic effects on the TM, leading to disease modifying IOP lowering therapies.

Original languageEnglish
Pages (from-to)849-859
Number of pages11
JournalInvestigative Ophthalmology and Visual Science
Volume58
Issue number2
DOIs
StatePublished - 1 Feb 2017

Fingerprint

Trabecular Meshwork
Proteins
Plasminogen Activator Inhibitor 1
Fibronectins
Bone and Bones
Inhibitor of Differentiation Protein 1
Extracellular Matrix
Bone Morphogenetic Protein Receptors
Plasminogen Inactivators
Messenger RNA
Bone Morphogenetic Proteins
Aqueous Humor
Extracellular Matrix Proteins
Human Activities
Fibrosis
Transcription Factors
Western Blotting
Immunohistochemistry

Keywords

  • BMP4
  • Fibronectin
  • ID1
  • ID3
  • TGF-β2
  • TM cells

Cite this

@article{26676192af0a46f7a68eb97f3383e4de,
title = "Role of ID proteins in BMP4 inhibition of profibrotic effects of TGF-β2 in human TM cells",
abstract = "PURPOSE. Increased expression of TGF-β2 in primary open-angle glaucoma (POAG) aqueous humor (AH) and trabecular meshwork (TM) causes deposition of extracellular matrix (ECM) in the TM and elevated IOP. Bone morphogenetic proteins (BMPs) regulate TGF-β2-induced ECM production. The underlying mechanism for BMP4 inhibition of TGF-β2-induced fibrosis remains undetermined. Bone morphogenic protein 4 induces inhibitor of DNA binding proteins (ID1, ID3), which suppress transcription factor activities to regulate gene expression. Our study will determine whether ID1and ID3 proteins are downstream targets of BMP4, which attenuates TGF-β2 induction of ECM proteins in TM cells. METHODS. Primary human TM cells were treated with BMP4, and ID1 and ID3 mRNA, and protein expression was determined by quantitative PCR (Q-PCR) and Western immunoblotting. Intracellular ID1 and ID3 protein localization was studied by immunocytochemistry. Transformed human TM cells (GTM3 cells) were transfected with ID1 or ID3 expression vectors to determine their potential inhibitory effects on TGF-β2-induced fibronectin and plasminogen activator inhibitor-I (PAI-1) protein expression. RESULTS. Basal expression of ID1-3 was detected in primary human TM cells. Bone morphogenic protein 4 significantly induced early expression of ID1 and ID3 mRNA (P < 0.05) and protein in primary TM cells, and a BMP receptor inhibitor blocked this induction. Overexpression of ID1 and ID3 significantly inhibited TGF-β2-induced expression of fibronectin and PAI-1 in TM cells (P < 0.01). CONCLUSIONS. Bone morphogenic protein 4 induced ID1 and ID3 expression suppresses TGFβ2 profibrotic activity in human TM cells. In the future, targeting specific regulators may control the TGF-β2 profibrotic effects on the TM, leading to disease modifying IOP lowering therapies.",
keywords = "BMP4, Fibronectin, ID1, ID3, TGF-β2, TM cells",
author = "Mody, {Avani A.} and Wordinger, {Robert J.} and Abbot Clark",
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language = "English",
volume = "58",
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journal = "Investigative Ophthalmology and Visual Science",
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Role of ID proteins in BMP4 inhibition of profibrotic effects of TGF-β2 in human TM cells. / Mody, Avani A.; Wordinger, Robert J.; Clark, Abbot.

In: Investigative Ophthalmology and Visual Science, Vol. 58, No. 2, 01.02.2017, p. 849-859.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Role of ID proteins in BMP4 inhibition of profibrotic effects of TGF-β2 in human TM cells

AU - Mody, Avani A.

AU - Wordinger, Robert J.

AU - Clark, Abbot

PY - 2017/2/1

Y1 - 2017/2/1

N2 - PURPOSE. Increased expression of TGF-β2 in primary open-angle glaucoma (POAG) aqueous humor (AH) and trabecular meshwork (TM) causes deposition of extracellular matrix (ECM) in the TM and elevated IOP. Bone morphogenetic proteins (BMPs) regulate TGF-β2-induced ECM production. The underlying mechanism for BMP4 inhibition of TGF-β2-induced fibrosis remains undetermined. Bone morphogenic protein 4 induces inhibitor of DNA binding proteins (ID1, ID3), which suppress transcription factor activities to regulate gene expression. Our study will determine whether ID1and ID3 proteins are downstream targets of BMP4, which attenuates TGF-β2 induction of ECM proteins in TM cells. METHODS. Primary human TM cells were treated with BMP4, and ID1 and ID3 mRNA, and protein expression was determined by quantitative PCR (Q-PCR) and Western immunoblotting. Intracellular ID1 and ID3 protein localization was studied by immunocytochemistry. Transformed human TM cells (GTM3 cells) were transfected with ID1 or ID3 expression vectors to determine their potential inhibitory effects on TGF-β2-induced fibronectin and plasminogen activator inhibitor-I (PAI-1) protein expression. RESULTS. Basal expression of ID1-3 was detected in primary human TM cells. Bone morphogenic protein 4 significantly induced early expression of ID1 and ID3 mRNA (P < 0.05) and protein in primary TM cells, and a BMP receptor inhibitor blocked this induction. Overexpression of ID1 and ID3 significantly inhibited TGF-β2-induced expression of fibronectin and PAI-1 in TM cells (P < 0.01). CONCLUSIONS. Bone morphogenic protein 4 induced ID1 and ID3 expression suppresses TGFβ2 profibrotic activity in human TM cells. In the future, targeting specific regulators may control the TGF-β2 profibrotic effects on the TM, leading to disease modifying IOP lowering therapies.

AB - PURPOSE. Increased expression of TGF-β2 in primary open-angle glaucoma (POAG) aqueous humor (AH) and trabecular meshwork (TM) causes deposition of extracellular matrix (ECM) in the TM and elevated IOP. Bone morphogenetic proteins (BMPs) regulate TGF-β2-induced ECM production. The underlying mechanism for BMP4 inhibition of TGF-β2-induced fibrosis remains undetermined. Bone morphogenic protein 4 induces inhibitor of DNA binding proteins (ID1, ID3), which suppress transcription factor activities to regulate gene expression. Our study will determine whether ID1and ID3 proteins are downstream targets of BMP4, which attenuates TGF-β2 induction of ECM proteins in TM cells. METHODS. Primary human TM cells were treated with BMP4, and ID1 and ID3 mRNA, and protein expression was determined by quantitative PCR (Q-PCR) and Western immunoblotting. Intracellular ID1 and ID3 protein localization was studied by immunocytochemistry. Transformed human TM cells (GTM3 cells) were transfected with ID1 or ID3 expression vectors to determine their potential inhibitory effects on TGF-β2-induced fibronectin and plasminogen activator inhibitor-I (PAI-1) protein expression. RESULTS. Basal expression of ID1-3 was detected in primary human TM cells. Bone morphogenic protein 4 significantly induced early expression of ID1 and ID3 mRNA (P < 0.05) and protein in primary TM cells, and a BMP receptor inhibitor blocked this induction. Overexpression of ID1 and ID3 significantly inhibited TGF-β2-induced expression of fibronectin and PAI-1 in TM cells (P < 0.01). CONCLUSIONS. Bone morphogenic protein 4 induced ID1 and ID3 expression suppresses TGFβ2 profibrotic activity in human TM cells. In the future, targeting specific regulators may control the TGF-β2 profibrotic effects on the TM, leading to disease modifying IOP lowering therapies.

KW - BMP4

KW - Fibronectin

KW - ID1

KW - ID3

KW - TGF-β2

KW - TM cells

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U2 - 10.1167/iovs.16-20472

DO - 10.1167/iovs.16-20472

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JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

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