title: Activated hepatic stellate cells are dependent on self-collagen, cleaved by membrane type 1 matrix metalloproteinase for their growth.
abstract: Stellate cells are distributed throughout organs, where, upon chronic damage, they become activated and proliferate to secrete collagen, which results in organ fibrosis. An intriguing property of hepatic stellate cells (HSCs) is that they undergo apoptosis when collagen is resolved by stopping tissue damage or by treatment, even though the mechanisms are unknown. Here we disclose the fact that HSCs, normal diploid cells, acquired dependence on collagen for their growth during the transition from quiescent to active states. The intramolecular RGD motifs of collagen were exposed by cleavage with their own membrane type 1 matrix metalloproteinase (MT1-MMP). The following evidence supports this conclusion. When rat activated HSCs (aHSCs) were transduced with siRNA against the collagen-specific chaperone gp46 to inhibit collagen secretion, the cells underwent autophagy followed by apoptosis. Concomitantly, the growth of aHSCs was suppressed, whereas that of quiescent HSCs was not. These in vitro results are compatible with the in vivo observation that apoptosis of aHSCs was induced in cirrhotic livers of rats treated with siRNAgp46. siRNA against MT1-MMP and addition of tissue inhibitor of metalloproteinase 2 (TIMP-2), which mainly inhibits MT1-MMP, also significantly suppressed the growth of aHSCs in vitro. The RGD inhibitors echistatin and GRGDS peptide and siRNA against the RGD receptor αVβ1 resulted in the inhibition of aHSCs growth. Transduction of siRNAs against gp46, αVβ1, and MT1-MMP to aHSCs inhibited the survival signal of PI3K/AKT/IκB. These results could provide novel antifibrosis strategies.
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Naoko Kubo
authors.initials: NK
authors.lastname: Birukawa
authors.affiliation: 4th Department of Internal Medicine, and.
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Murase
authors.affiliation: 4th Department of Internal Medicine, and.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Sato
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Akemi
authors.initials: A
authors.lastname: Kosaka
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Yoneda
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Nishita
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Fujita
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Miyuki
authors.initials: M
authors.lastname: Nishimura
authors.affiliation: Department of Basic Medical Science Department of Anatomy (1), Sapporo Medical University School of Medicine, 060-8556 Sapporo, Japan.
authors.forename: Takafumi
authors.initials: T
authors.lastname: Ninomiya
authors.affiliation: From the Department of Molecular Target Exploration, the Translational Research Group, Hokkaido Laboratory, Molecular Therapeutics Department, Corporate Business Development Division, Nitto Denko Corporation, Hokkaido, 001-0021 Sapporo, Japan, and.
authors.forename: Keiko
authors.initials: K
authors.lastname: Kajiwara
authors.affiliation: From the Department of Molecular Target Exploration, the Translational Research Group, Hokkaido Laboratory, Molecular Therapeutics Department, Corporate Business Development Division, Nitto Denko Corporation, Hokkaido, 001-0021 Sapporo, Japan, and.
authors.forename: Miyono
authors.initials: M
authors.lastname: Miyazaki
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Nakashima
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Sigenori
authors.initials: S
authors.lastname: Ota
authors.affiliation: From the Department of Molecular Target Exploration.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Murakami
authors.affiliation: From the Department of Molecular Target Exploration, the Translational Research Group, Hokkaido Laboratory, Molecular Therapeutics Department, Corporate Business Development Division, Nitto Denko Corporation, Hokkaido, 001-0021 Sapporo, Japan, and.
authors.forename: Yasunobu
authors.initials: Y
authors.lastname: Tanaka
authors.affiliation: From the Department of Molecular Target Exploration, the Translational Research Group, Hokkaido Laboratory, Molecular Therapeutics Department, Corporate Business Development Division, Nitto Denko Corporation, Hokkaido, 001-0021 Sapporo, Japan, and.
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Minomi
authors.affiliation: the Faculty of Advanced Life Science, Hokkaido University, 001-0021 Sapporo, Japan.
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Tamura
authors.affiliation: From the Department of Molecular Target Exploration, niitsu@sapmed.ac.jp.
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Niitsu
mesh_terms: D000818:Animals; D017209:Apoptosis; D049109:Cell Proliferation; D002470:Cell Survival; D003094:Collagen / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D050958:HSP47 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D055166:Hepatic Stellate Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D020672:I-kappa B Proteins / Q000378:metabolism; D016023:Integrins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D008103:Liver Cirrhosis / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology; D051379:Mice; D009842:Oligopeptides / Q000737:chemistry; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D003094:Collagen; D050958:HSP47 Heat-Shock Proteins; D020672:I-kappa B Proteins; D016023:Integrins; D053511:Matrix Metalloproteinase 14; D061965:Matrix Metalloproteinase Inhibitors; D009842:Oligopeptides; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D034741:RNA, Small Interfering; D019715:Tissue Inhibitor of Metalloproteinase-1; C494662:Serpinh1 protein, rat; C576839:TIMP1 protein, rat; C047981:arginyl-glycyl-aspartic acid
keywords: Apoptosis; Extracellular Matrix; Fibrosis; Heat Shock Protein (HSP); Integrin