Inhibition of type I collagen gene expression in normal and systemic sclerosis fibroblasts by a specific inhibitor of geranylgeranyl transferase I

Joel Rosenbloom, Biagio Saitta, Svetlana Gaidarova, Nora Sandorfi, Joan C. Rosenbloom, William R. Abrams, Andrew Hamilton, Said M. Sebti, Umberto Kucich, Sergio A. Jimenez

Research output: Contribution to journalArticle

Abstract

Objective. To examine the effects of specific inhibition of geranylgeranyl transferase I on the expression of types I and III collagen genes in normal and systemic sclerosis (SSc) dermal fibroblasts in vitro. Methods. Fibroblasts from 2 normal subjects and 4 SSc patients were incubated with 2-10 μM of GGTI-298, a specific geranylgeranyl transferase inhibitor. Type I collagen and fibronectin production were determined by enzyme-linked immunosorbent assay. Steady-state messenger RNA (mRNA) levels for α 1/4 (I), α2(I), and α1(III) collagens and fibronectin were assessed by Northern hybridization, and the transcription of the α1(I) collagen gene was examined by transient transfections with a reporter construct containing -5.3 kb of the gene. Results. GGTI-298 caused a dose-dependent inhibition of type I collagen production and a reduction in the steady-state levels of α1(I), α2(I), and α1(III) mRNA in normal and SSc cells. A 60-70% inhibition of type I collagen production and a 70-80% reduction in the mRNA levels for α1(I), α2(I), and α1(III) were observed at 10 μM GGTI-298. In contrast, the expression of fibronectin, cyclooxygenase 1, and GAPDH was not affected. The effects on α1(I) collagen mRNA resulted from a profound reduction in transcription of the α1(I) collagen gene promoter. GGTI-298 did not affect cellular viability or morphology. Conclusion. These results demonstrate that specific inhibition of geranylgeranyl prenylation causes a potent and selective inhibition of expression of the genes encoding types I and III collagens, without affecting cellular viability. The findings indicate that inhibition of geranylgeranyl prenylation should be further studied as a potential therapeutic approach for SSc and other fibrosing diseases.

Original languageEnglish (US)
Pages (from-to)1624-1232
Number of pages393
JournalArthritis and Rheumatism
Volume43
Issue number7
DOIs
StatePublished - Jul 2000

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Systemic Scleroderma
Transferases
Collagen Type I
Fibroblasts
Gene Expression
Collagen
Fibronectins
Prenylation
Messenger RNA
Collagen Type III
Genes
Cyclooxygenase 1
Transfection
Enzyme-Linked Immunosorbent Assay
Skin
GGTI 298

ASJC Scopus subject areas

  • Immunology
  • Rheumatology

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Inhibition of type I collagen gene expression in normal and systemic sclerosis fibroblasts by a specific inhibitor of geranylgeranyl transferase I. / Rosenbloom, Joel; Saitta, Biagio; Gaidarova, Svetlana; Sandorfi, Nora; Rosenbloom, Joan C.; Abrams, William R.; Hamilton, Andrew; Sebti, Said M.; Kucich, Umberto; Jimenez, Sergio A.

In: Arthritis and Rheumatism, Vol. 43, No. 7, 07.2000, p. 1624-1232.

Research output: Contribution to journalArticle

Rosenbloom, Joel ; Saitta, Biagio ; Gaidarova, Svetlana ; Sandorfi, Nora ; Rosenbloom, Joan C. ; Abrams, William R. ; Hamilton, Andrew ; Sebti, Said M. ; Kucich, Umberto ; Jimenez, Sergio A. / Inhibition of type I collagen gene expression in normal and systemic sclerosis fibroblasts by a specific inhibitor of geranylgeranyl transferase I. In: Arthritis and Rheumatism. 2000 ; Vol. 43, No. 7. pp. 1624-1232.
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abstract = "Objective. To examine the effects of specific inhibition of geranylgeranyl transferase I on the expression of types I and III collagen genes in normal and systemic sclerosis (SSc) dermal fibroblasts in vitro. Methods. Fibroblasts from 2 normal subjects and 4 SSc patients were incubated with 2-10 μM of GGTI-298, a specific geranylgeranyl transferase inhibitor. Type I collagen and fibronectin production were determined by enzyme-linked immunosorbent assay. Steady-state messenger RNA (mRNA) levels for α 1/4 (I), α2(I), and α1(III) collagens and fibronectin were assessed by Northern hybridization, and the transcription of the α1(I) collagen gene was examined by transient transfections with a reporter construct containing -5.3 kb of the gene. Results. GGTI-298 caused a dose-dependent inhibition of type I collagen production and a reduction in the steady-state levels of α1(I), α2(I), and α1(III) mRNA in normal and SSc cells. A 60-70{\%} inhibition of type I collagen production and a 70-80{\%} reduction in the mRNA levels for α1(I), α2(I), and α1(III) were observed at 10 μM GGTI-298. In contrast, the expression of fibronectin, cyclooxygenase 1, and GAPDH was not affected. The effects on α1(I) collagen mRNA resulted from a profound reduction in transcription of the α1(I) collagen gene promoter. GGTI-298 did not affect cellular viability or morphology. Conclusion. These results demonstrate that specific inhibition of geranylgeranyl prenylation causes a potent and selective inhibition of expression of the genes encoding types I and III collagens, without affecting cellular viability. The findings indicate that inhibition of geranylgeranyl prenylation should be further studied as a potential therapeutic approach for SSc and other fibrosing diseases.",
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T1 - Inhibition of type I collagen gene expression in normal and systemic sclerosis fibroblasts by a specific inhibitor of geranylgeranyl transferase I

AU - Rosenbloom, Joel

AU - Saitta, Biagio

AU - Gaidarova, Svetlana

AU - Sandorfi, Nora

AU - Rosenbloom, Joan C.

AU - Abrams, William R.

AU - Hamilton, Andrew

AU - Sebti, Said M.

AU - Kucich, Umberto

AU - Jimenez, Sergio A.

PY - 2000/7

Y1 - 2000/7

N2 - Objective. To examine the effects of specific inhibition of geranylgeranyl transferase I on the expression of types I and III collagen genes in normal and systemic sclerosis (SSc) dermal fibroblasts in vitro. Methods. Fibroblasts from 2 normal subjects and 4 SSc patients were incubated with 2-10 μM of GGTI-298, a specific geranylgeranyl transferase inhibitor. Type I collagen and fibronectin production were determined by enzyme-linked immunosorbent assay. Steady-state messenger RNA (mRNA) levels for α 1/4 (I), α2(I), and α1(III) collagens and fibronectin were assessed by Northern hybridization, and the transcription of the α1(I) collagen gene was examined by transient transfections with a reporter construct containing -5.3 kb of the gene. Results. GGTI-298 caused a dose-dependent inhibition of type I collagen production and a reduction in the steady-state levels of α1(I), α2(I), and α1(III) mRNA in normal and SSc cells. A 60-70% inhibition of type I collagen production and a 70-80% reduction in the mRNA levels for α1(I), α2(I), and α1(III) were observed at 10 μM GGTI-298. In contrast, the expression of fibronectin, cyclooxygenase 1, and GAPDH was not affected. The effects on α1(I) collagen mRNA resulted from a profound reduction in transcription of the α1(I) collagen gene promoter. GGTI-298 did not affect cellular viability or morphology. Conclusion. These results demonstrate that specific inhibition of geranylgeranyl prenylation causes a potent and selective inhibition of expression of the genes encoding types I and III collagens, without affecting cellular viability. The findings indicate that inhibition of geranylgeranyl prenylation should be further studied as a potential therapeutic approach for SSc and other fibrosing diseases.

AB - Objective. To examine the effects of specific inhibition of geranylgeranyl transferase I on the expression of types I and III collagen genes in normal and systemic sclerosis (SSc) dermal fibroblasts in vitro. Methods. Fibroblasts from 2 normal subjects and 4 SSc patients were incubated with 2-10 μM of GGTI-298, a specific geranylgeranyl transferase inhibitor. Type I collagen and fibronectin production were determined by enzyme-linked immunosorbent assay. Steady-state messenger RNA (mRNA) levels for α 1/4 (I), α2(I), and α1(III) collagens and fibronectin were assessed by Northern hybridization, and the transcription of the α1(I) collagen gene was examined by transient transfections with a reporter construct containing -5.3 kb of the gene. Results. GGTI-298 caused a dose-dependent inhibition of type I collagen production and a reduction in the steady-state levels of α1(I), α2(I), and α1(III) mRNA in normal and SSc cells. A 60-70% inhibition of type I collagen production and a 70-80% reduction in the mRNA levels for α1(I), α2(I), and α1(III) were observed at 10 μM GGTI-298. In contrast, the expression of fibronectin, cyclooxygenase 1, and GAPDH was not affected. The effects on α1(I) collagen mRNA resulted from a profound reduction in transcription of the α1(I) collagen gene promoter. GGTI-298 did not affect cellular viability or morphology. Conclusion. These results demonstrate that specific inhibition of geranylgeranyl prenylation causes a potent and selective inhibition of expression of the genes encoding types I and III collagens, without affecting cellular viability. The findings indicate that inhibition of geranylgeranyl prenylation should be further studied as a potential therapeutic approach for SSc and other fibrosing diseases.

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