Transforming growth factor-β2 induces expression of biologically active bone morphogenetic protein-1 in human trabecular meshwork cells

Tara Tovar-Vidales, Ashley M. Fitzgerald, Abbot Clark, Robert J. Wordinger

Research output: Contribution to journalArticleResearchpeer-review

12 Citations (Scopus)

Abstract

PURPOSE. There are limited studies on the factors that regulate the processing of TGF-β2 and extracellular matrix (ECM) proteins into their mature form. Bone morphogenic protein 1 (BMP1) is an enzyme responsible for the cleavage and maturation of growth factors and ECM proteins. The purpose of our study was to determine whether cultured human trabecular meshwork (TM) cells express BMP1, BMP1 expression is regulated by TGF-β2, BMP1 is biologically active, and BMP1 regulates LOX activity. METHODS. Primary human TM cells were isolated and subjected to quantitative PCR (qPCR) and Western immunoblotting (WB) for BMP1. BMP1 immunolocalization was performed in TM tissues. qPCR was used to determine BMP1 mRNA expression and WB results were used to determine BMP1 protein expression. BMP1 activity was measured in TM cells treated with TGF-β2 or with a combination of TGF-β2/UK383367. Lysyl oxidase (LOX) enzyme activity was evaluated by WB in TM cells treated with BMP1 or with a combination of BMP1/βaminoprorionitrile (BAPN). RESULTS. Human TM cells expressed mRNA and protein for BMP1. Exogenous TGF-β2 increased mRNA expression compared to their controls (P < 0.05). An ELISA showed TGF-β2- induced BMP1 secretion compared to their controls in all cell strains (P < 0.05). Secreted BMP1 stimulated LOX enzymatic activity in TM cells. CONCLUSIONS. BMP1 is expressed in the human TM. TGF-β2 induction of BMP1 may be responsible for increased processing of growth factors and ECM proteins into their mature forms, resulting in TM stiffness and resistance to ECM degradation.

Original languageEnglish
Pages (from-to)4741-4748
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume54
Issue number7
DOIs
StatePublished - 23 Jul 2013

Fingerprint

Trabecular Meshwork
Transforming Growth Factors
Bone and Bones
Proteins
Protein-Lysine 6-Oxidase
Extracellular Matrix Proteins
human BMP1 protein
Messenger RNA
Intercellular Signaling Peptides and Proteins
Polymerase Chain Reaction

Keywords

  • Bone morphogenetic protein 1
  • Trabecular meshwork
  • Transforming growth factor beta 2

Cite this

@article{a0d41787f7d848f2b1fd6595f9186069,
title = "Transforming growth factor-β2 induces expression of biologically active bone morphogenetic protein-1 in human trabecular meshwork cells",
abstract = "PURPOSE. There are limited studies on the factors that regulate the processing of TGF-β2 and extracellular matrix (ECM) proteins into their mature form. Bone morphogenic protein 1 (BMP1) is an enzyme responsible for the cleavage and maturation of growth factors and ECM proteins. The purpose of our study was to determine whether cultured human trabecular meshwork (TM) cells express BMP1, BMP1 expression is regulated by TGF-β2, BMP1 is biologically active, and BMP1 regulates LOX activity. METHODS. Primary human TM cells were isolated and subjected to quantitative PCR (qPCR) and Western immunoblotting (WB) for BMP1. BMP1 immunolocalization was performed in TM tissues. qPCR was used to determine BMP1 mRNA expression and WB results were used to determine BMP1 protein expression. BMP1 activity was measured in TM cells treated with TGF-β2 or with a combination of TGF-β2/UK383367. Lysyl oxidase (LOX) enzyme activity was evaluated by WB in TM cells treated with BMP1 or with a combination of BMP1/βaminoprorionitrile (BAPN). RESULTS. Human TM cells expressed mRNA and protein for BMP1. Exogenous TGF-β2 increased mRNA expression compared to their controls (P < 0.05). An ELISA showed TGF-β2- induced BMP1 secretion compared to their controls in all cell strains (P < 0.05). Secreted BMP1 stimulated LOX enzymatic activity in TM cells. CONCLUSIONS. BMP1 is expressed in the human TM. TGF-β2 induction of BMP1 may be responsible for increased processing of growth factors and ECM proteins into their mature forms, resulting in TM stiffness and resistance to ECM degradation.",
keywords = "Bone morphogenetic protein 1, Trabecular meshwork, Transforming growth factor beta 2",
author = "Tara Tovar-Vidales and Fitzgerald, {Ashley M.} and Abbot Clark and Wordinger, {Robert J.}",
year = "2013",
month = "7",
day = "23",
doi = "10.1167/iovs.13-12203",
language = "English",
volume = "54",
pages = "4741--4748",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "7",

}

Transforming growth factor-β2 induces expression of biologically active bone morphogenetic protein-1 in human trabecular meshwork cells. / Tovar-Vidales, Tara; Fitzgerald, Ashley M.; Clark, Abbot; Wordinger, Robert J.

In: Investigative Ophthalmology and Visual Science, Vol. 54, No. 7, 23.07.2013, p. 4741-4748.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Transforming growth factor-β2 induces expression of biologically active bone morphogenetic protein-1 in human trabecular meshwork cells

AU - Tovar-Vidales, Tara

AU - Fitzgerald, Ashley M.

AU - Clark, Abbot

AU - Wordinger, Robert J.

PY - 2013/7/23

Y1 - 2013/7/23

N2 - PURPOSE. There are limited studies on the factors that regulate the processing of TGF-β2 and extracellular matrix (ECM) proteins into their mature form. Bone morphogenic protein 1 (BMP1) is an enzyme responsible for the cleavage and maturation of growth factors and ECM proteins. The purpose of our study was to determine whether cultured human trabecular meshwork (TM) cells express BMP1, BMP1 expression is regulated by TGF-β2, BMP1 is biologically active, and BMP1 regulates LOX activity. METHODS. Primary human TM cells were isolated and subjected to quantitative PCR (qPCR) and Western immunoblotting (WB) for BMP1. BMP1 immunolocalization was performed in TM tissues. qPCR was used to determine BMP1 mRNA expression and WB results were used to determine BMP1 protein expression. BMP1 activity was measured in TM cells treated with TGF-β2 or with a combination of TGF-β2/UK383367. Lysyl oxidase (LOX) enzyme activity was evaluated by WB in TM cells treated with BMP1 or with a combination of BMP1/βaminoprorionitrile (BAPN). RESULTS. Human TM cells expressed mRNA and protein for BMP1. Exogenous TGF-β2 increased mRNA expression compared to their controls (P < 0.05). An ELISA showed TGF-β2- induced BMP1 secretion compared to their controls in all cell strains (P < 0.05). Secreted BMP1 stimulated LOX enzymatic activity in TM cells. CONCLUSIONS. BMP1 is expressed in the human TM. TGF-β2 induction of BMP1 may be responsible for increased processing of growth factors and ECM proteins into their mature forms, resulting in TM stiffness and resistance to ECM degradation.

AB - PURPOSE. There are limited studies on the factors that regulate the processing of TGF-β2 and extracellular matrix (ECM) proteins into their mature form. Bone morphogenic protein 1 (BMP1) is an enzyme responsible for the cleavage and maturation of growth factors and ECM proteins. The purpose of our study was to determine whether cultured human trabecular meshwork (TM) cells express BMP1, BMP1 expression is regulated by TGF-β2, BMP1 is biologically active, and BMP1 regulates LOX activity. METHODS. Primary human TM cells were isolated and subjected to quantitative PCR (qPCR) and Western immunoblotting (WB) for BMP1. BMP1 immunolocalization was performed in TM tissues. qPCR was used to determine BMP1 mRNA expression and WB results were used to determine BMP1 protein expression. BMP1 activity was measured in TM cells treated with TGF-β2 or with a combination of TGF-β2/UK383367. Lysyl oxidase (LOX) enzyme activity was evaluated by WB in TM cells treated with BMP1 or with a combination of BMP1/βaminoprorionitrile (BAPN). RESULTS. Human TM cells expressed mRNA and protein for BMP1. Exogenous TGF-β2 increased mRNA expression compared to their controls (P < 0.05). An ELISA showed TGF-β2- induced BMP1 secretion compared to their controls in all cell strains (P < 0.05). Secreted BMP1 stimulated LOX enzymatic activity in TM cells. CONCLUSIONS. BMP1 is expressed in the human TM. TGF-β2 induction of BMP1 may be responsible for increased processing of growth factors and ECM proteins into their mature forms, resulting in TM stiffness and resistance to ECM degradation.

KW - Bone morphogenetic protein 1

KW - Trabecular meshwork

KW - Transforming growth factor beta 2

UR - http://www.scopus.com/inward/record.url?scp=84880283394&partnerID=8YFLogxK

U2 - 10.1167/iovs.13-12203

DO - 10.1167/iovs.13-12203

M3 - Article

VL - 54

SP - 4741

EP - 4748

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 7

ER -