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PMID: 15987742 Published · ppublish English Comparative Study Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Smad2 and Smad3 play different roles in rat hepatic stellate cell function and alpha-smooth muscle actin organization.

Molecular biology of the cell ·Vol. 16 ·No. 9 ·2005-09-00 ·Pages 4214-24

Uemura M, Swenson ES, Gaça MD, Giordano FJ, Reiss M, Wells RG

Abstract

Hepatic stellate cells (HSC) play a central role in the pathogenesis of liver fibrosis, transdifferentiating in chronic liver disease from "quiescent" HSC to fibrogenic myofibroblasts. Transforming growth factor-beta (TGF-beta), acting both directly and indirectly, is a critical mediator of this process. To characterize the function of the TGF-beta signaling intermediates Smad2 and Smad3 in HSC, we infected primary rat HSC in culture with adenoviruses expressing wild-type and dominant negative Smads 2 and 3. Smad3-overexpressing cells exhibited increased deposition of fibronectin and type 1 collagen, increased chemotaxis, and decreased proliferation compared with uninfected cells and those infected with Smad2 or either dominant negative, demonstrating different biological functions for the two Smads. Additionally, coinfection experiments suggested that Smad2 and Smad3 signal via independent pathways. Smad3-overexpressing cells as well as TGF-beta-treated cells demonstrated more focal adhesions and increased alpha-smooth muscle actin (alpha-SMA) organization in stress fibers, although all cells reached the same level of alpha-SMA expression, indicating that Smad3 also regulates cytoskeletal organization in HSC. We suggest that TGF-beta, signaling via Smad3, plays an important role in the morphological and functional maturation of hepatic myofibroblasts.

MeSH Terms
Actins/metabolism Adenoviridae Animals Cell Proliferation Cells, Cultured Chemotaxis/physiology Extracellular Matrix Proteins/biosynthesis,genetics Genetic Vectors Liver/cytology,metabolism Male Rats Rats, Sprague-Dawley Stem Cells/cytology Transduction, Genetic Up-Regulation/physiology
Chemicals
Actins Extracellular Matrix Proteins smooth muscle actin, rat
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Uemura Masayuki
The University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
Swenson E Scott
Gaça Marianna D A
Giordano Frank J
Reiss Michael
Wells Rebecca G
References (53)
53 references, click to expand
  1. Lipocytes from normal rat liver release a neutral metalloproteinase that degrades basement membrane (type IV) collagen.
    J Clin Invest. 1989 Oct;84(4):1076-85 PMID: 2551922
  2. Maintenance of differentiated phenotype of cultured rat hepatic lipocytes by basement membrane matrix.
    J Biol Chem. 1989 Jun 25;264(18):10756-62 PMID: 2732246
  3. Transforming growth factors beta 1 and alpha in chronic liver disease. Effects of interferon alfa therapy.
    N Engl J Med. 1991 Apr 4;324(14):933-40 PMID: 1900574
  4. Isolated hepatic lipocytes and Kupffer cells from normal human liver: morphological and functional characteristics in primary culture.
    Hepatology. 1992 Feb;15(2):234-43 PMID: 1735526
  5. Transforming growth factor-beta 1 induces alpha-smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent and growing cultured fibroblasts.
    J Cell Biol. 1993 Jul;122(1):103-11 PMID: 8314838
  6. Use of recombinant adenovirus for metabolic engineering of mammalian cells.
    Methods Cell Biol. 1994;43 Pt A:161-89 PMID: 7823861
  7. Hepatic expression of mature transforming growth factor beta 1 in transgenic mice results in multiple tissue lesions.
    Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2572-6 PMID: 7708687
  8. Phosphorylation of Ser465 and Ser467 in the C terminus of Smad2 mediates interaction with Smad4 and is required for transforming growth factor-beta signaling.
    J Biol Chem. 1997 Oct 31;272(44):28107-15 PMID: 9346966
  9. TGF-beta signalling from cell membrane to nucleus through SMAD proteins.
    Nature. 1997 Dec 4;390(6659):465-71 PMID: 9393997
  10. Failure of egg cylinder elongation and mesoderm induction in mouse embryos lacking the tumor suppressor smad2.
    Proc Natl Acad Sci U S A. 1998 Aug 4;95(16):9378-83 PMID: 9689088
  11. Smads: transcriptional activators of TGF-beta responses.
    Cell. 1998 Dec 11;95(6):737-40 PMID: 9865691
  12. The role of TGFbeta1 in initiating hepatic stellate cell activation in vivo.
    J Hepatol. 1999 Jan;30(1):77-87 PMID: 9927153
  13. Blockade of type beta transforming growth factor signaling prevents liver fibrosis and dysfunction in the rat.
    Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2345-9 PMID: 10051644
  14. Targeted disruption of SMAD3 results in impaired mucosal immunity and diminished T cell responsiveness to TGF-beta.
    EMBO J. 1999 Mar 1;18(5):1280-91 PMID: 10064594
  15. Targeted disruption of Smad3 reveals an essential role in transforming growth factor beta-mediated signal transduction.
    Mol Cell Biol. 1999 Apr;19(4):2495-504 PMID: 10082515
  16. Role of transforming growth factor beta type II receptor in hepatic fibrosis: studies of human chronic hepatitis C and experimental fibrosis in rats.
    Hepatology. 1999 Jun;29(6):1730-8 PMID: 10347115
  17. Roles of autocrine TGF-beta receptor and Smad signaling in adipocyte differentiation.
    J Cell Biol. 2000 May 1;149(3):667-82 PMID: 10791980
  18. Fibrogenesis. V. TGF-beta signaling pathways.
    Am J Physiol Gastrointest Liver Physiol. 2000 Nov;279(5):G845-50 PMID: 11052979
  19. Identification of novel TGF-beta /Smad gene targets in dermal fibroblasts using a combined cDNA microarray/promoter transactivation approach.
    J Biol Chem. 2001 May 18;276(20):17058-62 PMID: 11279127
  20. Functional characterization of transforming growth factor beta signaling in Smad2- and Smad3-deficient fibroblasts.
    J Biol Chem. 2001 Jun 8;276(23):19945-53 PMID: 11262418
  21. The role of Smad3 in mediating mouse hepatic stellate cell activation.
    Hepatology. 2001 Jul;34(1):89-100 PMID: 11431738
  22. Transforming growth factor-beta repression of matrix metalloproteinase-1 in dermal fibroblasts involves Smad3.
    J Biol Chem. 2001 Oct 19;276(42):38502-10 PMID: 11502752
  23. Transforming growth factor beta-1 stimulates invasivity of hepatic stellate cells by engagement of the cell-associated fibrinolytic system.
    Growth Factors. 2001;19(2):87-100 PMID: 11769974
  24. Smad3 deficiency attenuates bleomycin-induced pulmonary fibrosis in mice.
    Am J Physiol Lung Cell Mol Physiol. 2002 Mar;282(3):L585-93 PMID: 11839555
  25. In vivo inhibition of rat stellate cell activation by soluble transforming growth factor beta type II receptor: a potential new therapy for hepatic fibrosis.
    Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12719-24 PMID: 10535989
  26. Mice lacking Smad3 show accelerated wound healing and an impaired local inflammatory response.
    Nat Cell Biol. 1999 Sep;1(5):260-6 PMID: 10559937
  27. A soluble transforming growth factor beta receptor expressed in muscle prevents liver fibrogenesis and dysfunction in rats.
    Hum Gene Ther. 2000 Jan 1;11(1):33-42 PMID: 10646637
  28. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury.
    J Biol Chem. 2000 Jan 28;275(4):2247-50 PMID: 10644669
  29. Transcriptional control by the TGF-beta/Smad signaling system.
    EMBO J. 2000 Apr 17;19(8):1745-54 PMID: 10775259
  30. Adenovirus-mediated gene transfer to nonparenchymal cells in normal and injured liver.
    Am J Physiol Gastrointest Liver Physiol. 2002 Mar;282(3):G565-72 PMID: 11842008
  31. Mice lacking Smad3 are protected against cutaneous injury induced by ionizing radiation.
    Am J Pathol. 2002 Mar;160(3):1057-68 PMID: 11891202
  32. Immortal activated human hepatic stellate cells generated by ectopic telomerase expression.
    Lab Invest. 2002 Mar;82(3):323-33 PMID: 11896211
  33. Roles of TGF-beta in hepatic fibrosis.
    Front Biosci. 2002 Apr 1;7:d793-807 PMID: 11897555
  34. Dose-dependent inhibition of hepatic fibrosis in mice by a TGF-beta soluble receptor: implications for antifibrotic therapy.
    Hepatology. 2002 May;35(5):1022-30 PMID: 11981752
  35. Myofibroblasts and mechano-regulation of connective tissue remodelling.
    Nat Rev Mol Cell Biol. 2002 May;3(5):349-63 PMID: 11988769
  36. Early endosomal regulation of Smad-dependent signaling in endothelial cells.
    J Biol Chem. 2002 May 17;277(20):18046-52 PMID: 11877415
  37. Adenoviral delivery of an antisense RNA complementary to the 3' coding sequence of transforming growth factor-beta1 inhibits fibrogenic activities of hepatic stellate cells.
    Cell Growth Differ. 2002 Jun;13(6):265-73 PMID: 12114216
  38. TGF beta receptor internalization into EEA1-enriched early endosomes: role in signaling to Smad2.
    J Cell Biol. 2002 Sep 30;158(7):1239-49 PMID: 12356868
  39. SARA, a FYVE domain protein, affects Rab5-mediated endocytosis.
    J Cell Sci. 2002 Dec 15;115(Pt 24):4755-63 PMID: 12432064
  40. Liver fibrosis: insights into migration of hepatic stellate cells in response to extracellular matrix and growth factors.
    Gastroenterology. 2003 Jan;124(1):147-59 PMID: 12512039
  41. Smad3 mediates the TGF-beta-induced contraction of type I collagen gels by mouse embryo fibroblasts.
    Cell Motil Cytoskeleton. 2003 Mar;54(3):248-53 PMID: 12589683
  42. Smads 2 and 3 are differentially activated by transforming growth factor-beta (TGF-beta ) in quiescent and activated hepatic stellate cells. Constitutive nuclear localization of Smads in activated cells is TGF-beta-independent.
    J Biol Chem. 2003 Mar 28;278(13):11721-8 PMID: 12547835
  43. Distinct endocytic pathways regulate TGF-beta receptor signalling and turnover.
    Nat Cell Biol. 2003 May;5(5):410-21 PMID: 12717440
  44. Alpha-smooth muscle actin is crucial for focal adhesion maturation in myofibroblasts.
    Mol Biol Cell. 2003 Jun;14(6):2508-19 PMID: 12808047
  45. The myofibroblast in wound healing and fibrocontractive diseases.
    J Pathol. 2003 Jul;200(4):500-3 PMID: 12845617
  46. Smad7 prevents activation of hepatic stellate cells and liver fibrosis in rats.
    Gastroenterology. 2003 Jul;125(1):178-91 PMID: 12851882
  47. Basement membrane-like matrix inhibits proliferation and collagen synthesis by activated rat hepatic stellate cells: evidence for matrix-dependent deactivation of stellate cells.
    Matrix Biol. 2003 May;22(3):229-39 PMID: 12853033
  48. Smads regulate collagen gel contraction by human dermal fibroblasts.
    Br J Dermatol. 2003 Sep;149(3):464-70 PMID: 14510976
  49. Distinct roles of Smad2-, Smad3-, and ERK-dependent pathways in transforming growth factor-beta1 regulation of pancreatic stellate cellular functions.
    J Biol Chem. 2004 Mar 5;279(10):8873-8 PMID: 14688282
  50. Smad3 as a mediator of the fibrotic response.
    Int J Exp Pathol. 2004 Apr;85(2):47-64 PMID: 15154911
  51. Proteomic profiling of bone marrow mesenchymal stem cells upon transforming growth factor beta1 stimulation.
    J Biol Chem. 2004 Oct 15;279(42):43725-34 PMID: 15302865
  52. Isolation and culture of hepatic lipocytes, Kupffer cells, and sinusoidal endothelial cells by density gradient centrifugation with Stractan.
    Anal Biochem. 1987 Feb 15;161(1):207-18 PMID: 3578783
  53. Cellular distribution of transforming growth factor-beta 1 and procollagen types I, III, and IV transcripts in carbon tetrachloride-induced rat liver fibrosis.
    J Clin Invest. 1990 Jun;85(6):1833-43 PMID: 1693377
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-02-0149
Published
2005-09-00
Epub
2005-00-29
Pages
4214-24
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1196331
Subset
IM
Grants
NIDDK NIH HHS · P30 DK050306 · United States
NIDDK NIH HHS · P30-DK050306 · United States
PHS HHS · R01-58123 · United States
External Links
PubMed source DOI: 10.1091/mbc.e05-02-0149 PMC Full Text View journal details
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