Inhibition of type I collagen gene expression in normal and systemic sclerosis fibroblasts by a specific inhibitor of geranylgeranyl transferase I.
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 microM 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 alpha1(I), alpha2(I), and alpha1(III) collagens and fibronectin were assessed by Northern hybridization, and the transcription of the alpha1(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 alpha1(I), alpha2(I), and alpha1(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 alpha1(I), alpha2(I), and alpha1(III) were observed at 10 microM GGTI-298. In contrast, the expression of fibronectin, cyclooxygenase 1, and GAPDH was not affected. The effects on alpha1(I) collagen mRNA resulted from a profound reduction in transcription of the alpha1(I) collagen gene promoter. GGTI-298 did not affect cellular viability or morphology.


CONCLUSIONS
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.
Rosenbloom
J
J
University of Pennsylvania School of Dental Medicine, Philadelphia, USA.
Saitta
B
B
Gaidarova
S
S
Sandorfi
N
N
Rosenbloom
J C
JC
Abrams
W R
WR
Hamilton
A D
AD
Sebti
S M
SM
Kucich
U
U
Jimenez
S A
SA
D000199:Actins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D019883:Alkyl and Aryl Transferases / Q000037:antagonists & inhibitors*; D001549:Benzamides / Q000494:pharmacology; D002460:Cell Line; D002461:Cell Line, Transformed; D003094:Collagen / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D051545:Cyclooxygenase 1; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000201:enzymology / Q000473:pathology; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008565:Membrane Proteins; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012595:Scleroderma, Systemic / Q000201:enzymology* / Q000473:pathology; D012867:Skin / Q000201:enzymology* / Q000473:pathology; D014158:Transcription, Genetic / Q000187:drug effects
D000199:Actins; D001549:Benzamides; D004791:Enzyme Inhibitors; D005353:Fibronectins; C102521:GGTI 298; D007527:Isoenzymes; D008565:Membrane Proteins; D012333:RNA, Messenger; D003094:Collagen; D051545:Cyclooxygenase 1; C496511:PTGS1 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D019883:Alkyl and Aryl Transferases; C083291:geranylgeranyltransferase type-I