Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis

Gautam Edhayan, Ray A. Ohara, W. Alex Stinson, M. Asif Amin, Takeo Isozaki, Christine M. Ha, G. Kenneth Haines, Rachel Morgan, Phillip L. Campbell, Ali Syed Arbab, Sean C. Friday, David A. Fox, Jeffrey H. Ruth

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background: Inhibitor of DNA binding 1 (Id1) is a nuclear protein containing a basic helix-loop-helix (bHLH) domain that regulates cell growth by selective binding and prevention of gene transcription. Sources of Id1 production in rheumatoid arthritis synovial tissue (RA ST) and its range of functional effects in RA remain to be clarified. Methods: We analyzed Id1 produced from synovial fibroblasts and endothelial cells (ECs) with histology and real-time polymerase chain reaction (RT-PCR). Fibroblast supernatants subjected to differential centrifugation to isolate and purify exosomes were measured for Id1 by enzyme-linked immunosorbent assay (ELISA). Western blotting of Id1-stimulated ECs was performed to determine the kinetics of intracellular protein phosphorylation. EC intracellular signaling pathways induced by Id1 were subsequently targeted with silencing RNA (siRNA) for angiogenesis inhibition. Results: By PCR and histologic analysis, we found that the primary source of Id1 in STs is from activated fibroblasts that correlate with inflammatory scores in human RA ST and in joints from K/BxN serum-induced mice. Normal (NL) and RA synovial fibroblasts increase Id1 production with stimulation by transforming growth factor beta (TGF-β). Most of the Id1 released by RA synovial fibroblasts is contained within exosomes. Endothelial progenitor cells (EPCs) and human dermal microvascular ECs (HMVECs) activate the Jnk signaling pathway in response to Id1, and Jnk siRNA reverses Id1-induced HMVEC vessel formation in Matrigel plugs in vivo. Conclusions: Id1 is a pleotropic molecule affecting angiogenesis, vasculogenesis, and fibrosis. Our data shows that Id1 is not only an important nuclear protein, but also can be released from fibroblasts via exosomes. The ability of extracellular Id1 to activate signaling pathways expands the role of Id1 in the orchestration of tissue inflammation.

Original languageEnglish (US)
Article number87
JournalArthritis Research and Therapy
Volume18
Issue number1
DOIs
StatePublished - Jan 1 2016
Externally publishedYes

Fingerprint

Rheumatoid Arthritis
Fibroblasts
DNA
Exosomes
Endothelial Cells
Nuclear Proteins
RNA Interference
Helix-Loop-Helix Motifs
Skin
Centrifugation
Transforming Growth Factor beta
Real-Time Polymerase Chain Reaction
Histology
Fibrosis
Joints
Western Blotting
Enzyme-Linked Immunosorbent Assay
Phosphorylation
Inflammation

Keywords

  • Angiogenesis
  • Fibroblasts
  • Inflammation
  • Inhibitor of DNA binding-1 protein (Id1)
  • Rheumatoid arthritis

ASJC Scopus subject areas

  • Rheumatology
  • Immunology and Allergy
  • Immunology

Cite this

Edhayan, G., Ohara, R. A., Alex Stinson, W., Asif Amin, M., Isozaki, T., Ha, C. M., ... Ruth, J. H. (2016). Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis. Arthritis Research and Therapy, 18(1), [87]. https://doi.org/10.1186/S13075-016-0984-3

Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis. / Edhayan, Gautam; Ohara, Ray A.; Alex Stinson, W.; Asif Amin, M.; Isozaki, Takeo; Ha, Christine M.; Kenneth Haines, G.; Morgan, Rachel; Campbell, Phillip L.; Arbab, Ali Syed; Friday, Sean C.; Fox, David A.; Ruth, Jeffrey H.

In: Arthritis Research and Therapy, Vol. 18, No. 1, 87, 01.01.2016.

Research output: Contribution to journalArticle

Edhayan, G, Ohara, RA, Alex Stinson, W, Asif Amin, M, Isozaki, T, Ha, CM, Kenneth Haines, G, Morgan, R, Campbell, PL, Arbab, AS, Friday, SC, Fox, DA & Ruth, JH 2016, 'Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis', Arthritis Research and Therapy, vol. 18, no. 1, 87. https://doi.org/10.1186/S13075-016-0984-3
Edhayan, Gautam ; Ohara, Ray A. ; Alex Stinson, W. ; Asif Amin, M. ; Isozaki, Takeo ; Ha, Christine M. ; Kenneth Haines, G. ; Morgan, Rachel ; Campbell, Phillip L. ; Arbab, Ali Syed ; Friday, Sean C. ; Fox, David A. ; Ruth, Jeffrey H. / Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis. In: Arthritis Research and Therapy. 2016 ; Vol. 18, No. 1.
@article{c1968f209237436497cf32dd65a7ad1f,
title = "Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis",
abstract = "Background: Inhibitor of DNA binding 1 (Id1) is a nuclear protein containing a basic helix-loop-helix (bHLH) domain that regulates cell growth by selective binding and prevention of gene transcription. Sources of Id1 production in rheumatoid arthritis synovial tissue (RA ST) and its range of functional effects in RA remain to be clarified. Methods: We analyzed Id1 produced from synovial fibroblasts and endothelial cells (ECs) with histology and real-time polymerase chain reaction (RT-PCR). Fibroblast supernatants subjected to differential centrifugation to isolate and purify exosomes were measured for Id1 by enzyme-linked immunosorbent assay (ELISA). Western blotting of Id1-stimulated ECs was performed to determine the kinetics of intracellular protein phosphorylation. EC intracellular signaling pathways induced by Id1 were subsequently targeted with silencing RNA (siRNA) for angiogenesis inhibition. Results: By PCR and histologic analysis, we found that the primary source of Id1 in STs is from activated fibroblasts that correlate with inflammatory scores in human RA ST and in joints from K/BxN serum-induced mice. Normal (NL) and RA synovial fibroblasts increase Id1 production with stimulation by transforming growth factor beta (TGF-β). Most of the Id1 released by RA synovial fibroblasts is contained within exosomes. Endothelial progenitor cells (EPCs) and human dermal microvascular ECs (HMVECs) activate the Jnk signaling pathway in response to Id1, and Jnk siRNA reverses Id1-induced HMVEC vessel formation in Matrigel plugs in vivo. Conclusions: Id1 is a pleotropic molecule affecting angiogenesis, vasculogenesis, and fibrosis. Our data shows that Id1 is not only an important nuclear protein, but also can be released from fibroblasts via exosomes. The ability of extracellular Id1 to activate signaling pathways expands the role of Id1 in the orchestration of tissue inflammation.",
keywords = "Angiogenesis, Fibroblasts, Inflammation, Inhibitor of DNA binding-1 protein (Id1), Rheumatoid arthritis",
author = "Gautam Edhayan and Ohara, {Ray A.} and {Alex Stinson}, W. and {Asif Amin}, M. and Takeo Isozaki and Ha, {Christine M.} and {Kenneth Haines}, G. and Rachel Morgan and Campbell, {Phillip L.} and Arbab, {Ali Syed} and Friday, {Sean C.} and Fox, {David A.} and Ruth, {Jeffrey H.}",
year = "2016",
month = "1",
day = "1",
doi = "10.1186/S13075-016-0984-3",
language = "English (US)",
volume = "18",
journal = "Arthritis Research and Therapy",
issn = "1478-6354",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Inflammatory properties of inhibitor of DNA binding 1 secreted by synovial fibroblasts in rheumatoid arthritis

AU - Edhayan, Gautam

AU - Ohara, Ray A.

AU - Alex Stinson, W.

AU - Asif Amin, M.

AU - Isozaki, Takeo

AU - Ha, Christine M.

AU - Kenneth Haines, G.

AU - Morgan, Rachel

AU - Campbell, Phillip L.

AU - Arbab, Ali Syed

AU - Friday, Sean C.

AU - Fox, David A.

AU - Ruth, Jeffrey H.

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Background: Inhibitor of DNA binding 1 (Id1) is a nuclear protein containing a basic helix-loop-helix (bHLH) domain that regulates cell growth by selective binding and prevention of gene transcription. Sources of Id1 production in rheumatoid arthritis synovial tissue (RA ST) and its range of functional effects in RA remain to be clarified. Methods: We analyzed Id1 produced from synovial fibroblasts and endothelial cells (ECs) with histology and real-time polymerase chain reaction (RT-PCR). Fibroblast supernatants subjected to differential centrifugation to isolate and purify exosomes were measured for Id1 by enzyme-linked immunosorbent assay (ELISA). Western blotting of Id1-stimulated ECs was performed to determine the kinetics of intracellular protein phosphorylation. EC intracellular signaling pathways induced by Id1 were subsequently targeted with silencing RNA (siRNA) for angiogenesis inhibition. Results: By PCR and histologic analysis, we found that the primary source of Id1 in STs is from activated fibroblasts that correlate with inflammatory scores in human RA ST and in joints from K/BxN serum-induced mice. Normal (NL) and RA synovial fibroblasts increase Id1 production with stimulation by transforming growth factor beta (TGF-β). Most of the Id1 released by RA synovial fibroblasts is contained within exosomes. Endothelial progenitor cells (EPCs) and human dermal microvascular ECs (HMVECs) activate the Jnk signaling pathway in response to Id1, and Jnk siRNA reverses Id1-induced HMVEC vessel formation in Matrigel plugs in vivo. Conclusions: Id1 is a pleotropic molecule affecting angiogenesis, vasculogenesis, and fibrosis. Our data shows that Id1 is not only an important nuclear protein, but also can be released from fibroblasts via exosomes. The ability of extracellular Id1 to activate signaling pathways expands the role of Id1 in the orchestration of tissue inflammation.

AB - Background: Inhibitor of DNA binding 1 (Id1) is a nuclear protein containing a basic helix-loop-helix (bHLH) domain that regulates cell growth by selective binding and prevention of gene transcription. Sources of Id1 production in rheumatoid arthritis synovial tissue (RA ST) and its range of functional effects in RA remain to be clarified. Methods: We analyzed Id1 produced from synovial fibroblasts and endothelial cells (ECs) with histology and real-time polymerase chain reaction (RT-PCR). Fibroblast supernatants subjected to differential centrifugation to isolate and purify exosomes were measured for Id1 by enzyme-linked immunosorbent assay (ELISA). Western blotting of Id1-stimulated ECs was performed to determine the kinetics of intracellular protein phosphorylation. EC intracellular signaling pathways induced by Id1 were subsequently targeted with silencing RNA (siRNA) for angiogenesis inhibition. Results: By PCR and histologic analysis, we found that the primary source of Id1 in STs is from activated fibroblasts that correlate with inflammatory scores in human RA ST and in joints from K/BxN serum-induced mice. Normal (NL) and RA synovial fibroblasts increase Id1 production with stimulation by transforming growth factor beta (TGF-β). Most of the Id1 released by RA synovial fibroblasts is contained within exosomes. Endothelial progenitor cells (EPCs) and human dermal microvascular ECs (HMVECs) activate the Jnk signaling pathway in response to Id1, and Jnk siRNA reverses Id1-induced HMVEC vessel formation in Matrigel plugs in vivo. Conclusions: Id1 is a pleotropic molecule affecting angiogenesis, vasculogenesis, and fibrosis. Our data shows that Id1 is not only an important nuclear protein, but also can be released from fibroblasts via exosomes. The ability of extracellular Id1 to activate signaling pathways expands the role of Id1 in the orchestration of tissue inflammation.

KW - Angiogenesis

KW - Fibroblasts

KW - Inflammation

KW - Inhibitor of DNA binding-1 protein (Id1)

KW - Rheumatoid arthritis

UR - http://www.scopus.com/inward/record.url?scp=85008659776&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85008659776&partnerID=8YFLogxK

U2 - 10.1186/S13075-016-0984-3

DO - 10.1186/S13075-016-0984-3

M3 - Article

C2 - 27071670

AN - SCOPUS:85008659776

VL - 18

JO - Arthritis Research and Therapy

JF - Arthritis Research and Therapy

SN - 1478-6354

IS - 1

M1 - 87

ER -