CXCL12 induces hepatic stellate cell contraction through a calcium-independent pathway.

Liver fibrosis, with subsequent development of cirrhosis and ultimately portal hypertension, results in the death of patients with end-stage liver disease if liver transplantation is not performed. Hepatic stellate cells (HSCs), central mediators of liver fibrosis, resemble tissue pericytes and regulate intrahepatic blood flow by modulating pericapillary resistance. Therefore, HSCs can contribute to portal hypertension in patients with chronic liver disease (CLD). We have previously demonstrated that activated HSCs express functional chemokine receptor, CXCR4, and that receptor engagement by its ligand, CXCL12, which is increased in patients with CLD, leads to further stellate cell activation in a CXCR4-specific manner. We therefore hypothesized that CXCL12 promotes HSC contraction in a CXCR4-dependent manner. Stimulation of HSCs on collagen gel lattices with CXCL12 led to gel contraction and myosin light chain (MLC) phosphorylation, which was blocked by addition of AMD3100, a CXCR4 small molecule inhibitor. These effects were further mediated by the Rho kinase pathway since both Rho kinase knockdown or Y-27632, a Rho kinase inhibitor, blocked CXCL12 induced phosphorylation of MLC and gel contraction. BAPTA-AM, a calcium chelator, had no effect, indicating that this pathway is calcium sensitive but not calcium dependent. In conclusion, CXCL12 promotes stellate cell contractility in a predominantly calcium-independent fashion. Our data demonstrates a novel role of CXCL12 in stellate cell contraction and the availability of small molecule inhibitors of the CXCL12/CXCR4 axis justifies further investigation into its potential as therapeutic target for portal hypertension.

[1]  H. Broxmeyer,et al.  CCR5 Ligands Modulate CXCL12-Induced Chemotaxis, Adhesion, and Akt Phosphorylation of Human Cord Blood CD34+ Cells1 , 2009, The Journal of Immunology.

[2]  Jeffrey B. Kopp,et al.  TGF- and fibrosis , 1999 .

[3]  Kazuhiko Yamamoto,et al.  Cytokine‐mediated regulation of CXCR4 expression in human neutrophils , 2002, Journal of leukocyte biology.

[4]  N. Hotchin,et al.  Distinct Roles for ROCK1 and ROCK2 in the Regulation of Keratinocyte Differentiation , 2009, PloS one.

[5]  Olivia T. Abar,et al.  Functional linkage of cirrhosis‐predictive single nucleotide polymorphisms of toll‐like receptor 4 to hepatic stellate cell responses , 2009, Hepatology.

[6]  S. Friedman Hepatic stellate cells: protean, multifunctional, and enigmatic cells of the liver. , 2008, Physiological reviews.

[7]  J. Prakash,et al.  Reduction of Fibrogenesis by Selective Delivery of a Rho Kinase Inhibitor to Hepatic Stellate Cells in Mice , 2011, Journal of Pharmacology and Experimental Therapeutics.

[8]  W. Streit,et al.  Transforming growth factor‐β1 increases CXCR4 expression, stromal‐derived factor‐1α‐stimulated signalling and human immunodeficiency virus‐1 entry in human monocyte‐derived macrophages , 2005 .

[9]  Ido D. Weiss,et al.  Involvement of the CXCL12/CXCR4 pathway in the advanced liver disease that is associated with hepatitis C virus or hepatitis B virus , 2004, European journal of immunology.

[10]  Jiaquan Xu,et al.  Deaths: preliminary data for 2009. , 2011, National vital statistics reports : from the Centers for Disease Control and Prevention, National Center for Health Statistics, National Vital Statistics System.

[11]  M. Pinzani The Hepatic Stellate Cell , 2008 .

[12]  Geoffrey C Gurtner,et al.  Progenitor cell trafficking is regulated by hypoxic gradients through HIF-1 induction of SDF-1 , 2004, Nature Medicine.

[13]  M. Dwinell,et al.  Calcium Mobilization Triggered by the Chemokine CXCL12 Regulates Migration in Wounded Intestinal Epithelial Monolayers* , 2010, The Journal of Biological Chemistry.

[14]  N. Kawada,et al.  The contraction of hepatic stellate (Ito) cells stimulated with vasoactive substances. Possible involvement of endothelin 1 and nitric oxide in the regulation of the sinusoidal tonus. , 1993, European journal of biochemistry.

[15]  D. Häussinger,et al.  The niche of stellate cells within rat liver , 2009, Hepatology.

[16]  Xi Liu‐DeRyke,et al.  Medical management of variceal hemorrhage. , 2010, Critical care nursing clinics of North America.

[17]  A. Blauvelt,et al.  Cytokines regulate expression and function of the HIV coreceptor CXCR4 on human mature dendritic cells. , 1998, Journal of immunology.

[18]  J. Prakash,et al.  Increased Liver Uptake and Reduced Hepatic Stellate Cell Activation with a Cell-Specific Conjugate of the Rho-kinase Inhibitor Y27632 , 2011, Pharmaceutical Research.

[19]  D. Sims,et al.  The pericyte--a review. , 1986, Tissue & cell.

[20]  N. Heveker,et al.  Monomeric and dimeric CXCL12 inhibit metastasis through distinct CXCR4 interactions and signaling pathways , 2011, Proceedings of the National Academy of Sciences.

[21]  H. Broxmeyer,et al.  Altered responsiveness to chemokines due to targeted disruption of SHIP. , 1999, The Journal of clinical investigation.

[22]  Ritu Agarwal,et al.  Hepatic stellate cells express functional CXCR4: Role in stromal cell–derived factor‐1α–mediated stellate cell activation , 2009, Hepatology.

[23]  J. Henriksen,et al.  [The hepatic stellate cell]. , 2002, Ugeskrift for laeger.

[24]  M. Iizuka,et al.  Increased contractility of hepatic stellate cells in cirrhosis is mediated by enhanced Ca2+-dependent and Ca2+-sensitization pathways. , 2011, American journal of physiology. Gastrointestinal and liver physiology.

[25]  R. Bataller,et al.  Angiotensin II induces contraction and proliferation of human hepatic stellate cells. , 2000, Gastroenterology.

[26]  D. Rockey,et al.  Endothelin induced contractility of stellate cells from normal and cirrhotic rat liver: implications for regulation of portal pressure and resistance , 1996, Hepatology.

[27]  F. Sellke,et al.  Human coronary microvascular effects of cardioplegia-induced stromal-derived factor-1alpha. , 2006, The Annals of thoracic surgery.

[28]  Ramon Bataller,et al.  Human hepatic stellate cells express CCR5 and RANTES to induce proliferation and migration. , 2003, American journal of physiology. Gastrointestinal and liver physiology.