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Author Notes:

Email: Elisabetta A Renzoni - e.renzoni@imperial.ac.uk; David J Abraham - d.abraham@rfc.ucl.ac.uk; Sarah Howat - sarah.howat@kcl.ac.uk; Xu Shi-Wen - shiwen@rfc.ucl.ac.uk; Piersante Sestini - sestini@unisi.it; George Bou-Gharios - e.renzoni@imperial.ac.uk; Athol U Wells - a.wells@rbh.nthames.nhs.uk; Srihari Veeraraghavan - srihari1@yahoo.com; Andrew G Nicholson a.nicholson@rbh.nthames.nhs.uk; Christopher P Denton - c.denton@rfc.ucl.ac.uk; Andrew Leask - a.leask@rfc.ucl.ac.uk; Jeremy D Pearson - jeremy.pearson@kcl.ac.uk; Carol M Black - c.black@rfc.ucl.ac.uk; Kenneth I Welsh - k.welsh@imperial.ac.uk; Roland M du Bois* - R.DuBois@rbh.nthames.nhs.uk

EAR participated in the design and interpretation of the study, carried out the cell culture work and participated in the microarray work, performed immunohistochemistry staining, and drafted the manuscript.

DJA participated in the design and coordination of the study and in the preparation of the manuscript, SH performed the RT-PCR assays, XSW carried out the Western Blot analysis, PS performed the statistical analysis and participated in the interpretation of results and preparation of the manuscript, GBG participated in the microarray work, AUW participated in the interpretation of results, SV participated in cell line selection and clinical characterization, AGN reviewed fibrotic lung biopsies and interpreted immunohistochemistry staining, CD and CMB contributed towards the overall organizational setup for the study of lung fibroblast lines and participated in the interpretation of results, AL and JDP participated in the preparation of the manuscript, KIW conceived of the study and participated in the design, RdB participated in study design, interpretation and coordination.

All authors read and approved the final manuscript.

We are grateful to Helen Causton, Laurence Game, Nicola Cooley and Helen Banks of the CSC/IC Microarray Centre for expert help with Affymetrix microarray experiments.

We thank Carmen Fonseca, Paul Beirne and Alan Holmes for their technical expertise and for their review of the manuscript.

Subjects:

Research Funding:

This work was supported by the Royal Brompton & Harefield Clinical Research Fund, by the Raynaud's and Scleroderma Association Trust, the Scleroderma Society, the Welton Foundation, the Rosetrees Charitable Trust, and by the Arthritis and Research Council.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Respiratory System
  • RESPIRATORY SYSTEM
  • GROWTH-FACTOR-BETA
  • IDIOPATHIC PULMONARY FIBROSIS
  • ANGIOTENSIN-II
  • CARDIAC FIBROBLASTS
  • SYSTEMIC-SCLEROSIS
  • TYPE-1 RECEPTOR
  • INHIBITOR
  • DISEASE
  • DEGRADATION
  • ACTIVATION
  • PULMONARY

Gene expression profiling reveals novel TGFβ targets in adult lung fibroblasts

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Journal Title:

Respiratory Research

Volume:

Volume 5, Number 24

Publisher:

, Pages 24-24

Type of Work:

Article | Final Publisher PDF

Abstract:

Background: Transforming growth factor beta (TGFβ), a multifunctional cytokine, plays a crucial role in the accumulation of extracellular matrix components in lung fibrosis, where lung fibroblasts are considered to play a major role. Even though the effects of TGFβ on the gene expression of several proteins have been investigated in several lung fibroblast cell lines, the global pattern of response to this cytokine in adult lung fibroblasts is still unknown. Methods: We used Affymetrix oligonucleotide microarrays U95v2, containing approximately 12,000 human genes, to study the transcriptional profile in response to a four hour treatment with TGFβ in control lung fibroblasts and in fibroblasts from patients with idiopathic and scleroderma-associated pulmonary fibrosis. A combination of the Affymetrix change algorithm (Microarray Suite 5) and of analysis of variance models was used to identify TGFβ-regulated genes. Additional criteria were an average up- or down- regulation of at least two fold. Results: Exposure of fibroblasts to TGFβ had a profound impact on gene expression, resulting in regulation of 129 transcripts. We focused on genes not previously found to be regulated by TGFβ in lung fibroblasts or other cell types, including nuclear co-repressor 2, SMAD specific E3 ubiquitin protein ligase 2 (SMURF2), bone morphogenetic protein 4, and angiotensin II receptor type 1 (AGTR1), and confirmed the microarray results by real time-PCR. Western Blotting confirmed induction at the protein level of AGTR1, the most highly induced gene in both control and fibrotic lung fibroblasts among genes encoding for signal transduction molecules. Upregulation of AGTR1 occurred through the MKK1/MKK2 signalling pathway. Immunohistochemical staining showed AGTR1 expression by lung fibroblasts in fibroblastic foci within biopsies of idiopathic pulmonary fibrosis. Conclusions: This study identifies several novel TGFβ targets in lung fibroblasts, and confirms with independent methods the induction of angiotensin II receptor type 1, underlining a potential role for angiotensin II receptor 1 antagonism in the treatment of lung fibrosis.

Copyright information:

© 2004 Renzoni et al; licensee BioMed Central Ltd.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 2.0 Generic License (http://creativecommons.org/licenses/by/2.0/).

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