Biologically-active laminin-111 fragment that modulates the epithelial-to-mesenchymal transition in embryonic stem cells

Significance Laminin-111 is one of the first extracellular matrix proteins expressed during embryogenesis and has been studied for decades, mainly because of its major role in assembling the basement membrane, but also because it has now become one of the most popular cell culture substrates for embryonic stem cells. However, considering the importance of this protein, the role laminin-111 plays during matrix remodeling—which is not only of great interest when seeking to understand cell–matrix interactions, but also when using laminin as a substrate for tissue engineering—is still unclear. Our findings propose a previously unidentified role of laminin-111 that goes beyond basement membrane assembly and an important involvement in the regulation of the epithelial-to-mesenchymal transition. The dynamic interplay between the extracellular matrix and embryonic stem cells (ESCs) constitutes one of the key steps in understanding stem cell differentiation in vitro. Here we report a biologically-active laminin-111 fragment generated by matrix metalloproteinase 2 (MMP2) processing, which is highly up-regulated during differentiation. We show that the β1-chain–derived fragment interacts via α3β1-integrins, thereby triggering the down-regulation of MMP2 in mouse and human ESCs. Additionally, the expression of MMP9 and E-cadherin is up-regulated in mouse ESCs—key players in the epithelial-to-mesenchymal transition. We also demonstrate that the fragment acts through the α3β1-integrin/extracellular matrix metalloproteinase inducer complex. This study reveals a previously unidentified role of laminin-111 in early stem cell differentiation that goes far beyond basement membrane assembly and a mechanism by which an MMP2-cleaved laminin fragment regulates the expression of E-cadherin, MMP2, and MMP9.

[1]  L. V. Van Laake,et al.  Recombinant Vitronectin Is a Functionally Defined Substrate That Supports Human Embryonic Stem Cell Self‐Renewal via αVβ5 Integrin , 2008, Stem cells.

[2]  P. Stern,et al.  E-cadherin inhibits cell surface localization of the pro-migratory 5T4 oncofetal antigen in mouse embryonic stem cells. , 2007, Molecular biology of the cell.

[3]  K. Sekiguchi,et al.  Laminin E8 fragments support efficient adhesion and expansion of dissociated human pluripotent stem cells , 2012, Nature Communications.

[4]  I. Campbell,et al.  Interdomain Tilt Angle Determines Integrin-dependent Function of the Ninth and Tenth FIII Domains of Human Fibronectin* , 2004, Journal of Biological Chemistry.

[5]  Vandana Iyer,et al.  α3β1 integrin regulates MMP-9 mRNA stability in immortalized keratinocytes: a novel mechanism of integrin-mediated MMP gene expression , 2005, Journal of Cell Science.

[6]  H. Mizuguchi,et al.  Long-Term Self-Renewal of Human ES/iPS-Derived Hepatoblast-like Cells on Human Laminin 111-Coated Dishes , 2013, Stem cell reports.

[7]  Takako Sasaki,et al.  Laminin: the crux of basement membrane assembly. , 2004, The Journal of cell biology.

[8]  N. Brown,et al.  Integrins in development: moving on, responding to, and sticking to the extracellular matrix. , 2002, Developmental cell.

[9]  J. Hescheler,et al.  Inhibition of Tumor-Induced Angiogenesis and Matrix-Metalloproteinase Expression in Confrontation Cultures of Embryoid Bodies and Tumor Spheroids by Plant Ingredients Used in Traditional Chinese Medicine , 2003, Laboratory Investigation.

[10]  M. Nakai,et al.  Expression of extracellular matrix metalloproteinase inducer and matrix metalloproteinases during mouse embryonic development. , 2007, Reproduction.

[11]  F. Berditchevski,et al.  Generation of Monoclonal Antibodies to Integrin-associated Proteins , 1997, The Journal of Biological Chemistry.

[12]  H. Guo,et al.  The human tumor cell-derived collagenase stimulatory factor (renamed EMMPRIN) is a member of the immunoglobulin superfamily. , 1995, Cancer research.

[13]  F. Berditchevski,et al.  Generation of monoclonal antibodies to integrin-associated proteins. Evidence that alpha3beta1 complexes with EMMPRIN/basigin/OX47/M6. , 1997, The Journal of biological chemistry.

[14]  G. Laurie,et al.  Exposure of Cryptic Domains in the α1-chain of Laminin-1 by Elastase Stimulates Macrophages Urokinase and Matrix Metalloproteinase-9 Expression* , 2002, The Journal of Biological Chemistry.

[15]  P. Lonai,et al.  Expression and biological role of laminin-1. , 2003, Matrix biology : journal of the International Society for Matrix Biology.

[16]  S. Carbonetto,et al.  Mapping of network-forming, heparin-binding, and alpha 1 beta 1 integrin-recognition sites within the alpha-chain short arm of laminin-1. , 1995, The Journal of biological chemistry.

[17]  Hai-yan Lin,et al.  Effects of E-cadherin on mouse embryo implantation and expression of matrix metalloproteinase-2 and -9. , 2006, Biochemical and biophysical research communications.

[18]  Ashley C. Brown,et al.  Directing epithelial to mesenchymal transition through engineered microenvironments displaying orthogonal adhesive and mechanical cues. , 2012, Journal of biomedical materials research. Part A.

[19]  J. Thiery,et al.  Complex networks orchestrate epithelial–mesenchymal transitions , 2006, Nature Reviews Molecular Cell Biology.

[20]  D. Edwards,et al.  The regulation of matrix metalloproteinases and their inhibitors. , 2008, The international journal of biochemistry & cell biology.

[21]  R. Huang,et al.  Epithelial-Mesenchymal Transitions in Development and Disease , 2009, Cell.

[22]  K. Tryggvason,et al.  Laminin‐511 but Not ‐332, ‐111, or ‐411 Enables Mouse Embryonic Stem Cell Self‐Renewal In Vitro , 2008, Stem cells.

[23]  K. Sekiguchi,et al.  Recombinant human laminin isoforms can support the undifferentiated growth of human embryonic stem cells. , 2008, Biochemical and biophysical research communications.

[24]  J. Kreidberg.  Functions of α3β1 integrin , 2000 .

[25]  V. Quaranta,et al.  Tales from the crypt[ic] sites of the extracellular matrix. , 2003, Trends in cell biology.

[26]  E. Hohenester,et al.  Laminin Network Formation Studied by Reconstitution of Ternary Nodes in Solution* , 2012, The Journal of Biological Chemistry.

[27]  Y. Seyama,et al.  Anti‐α3 integrin antibody induces the activated form of matrix metalloprotease‐2 (MMP‐2) with concomitant stimulation of invasion through matrigel by human rhabdomyosarcoma cells , 1997, International journal of cancer.

[28]  D. Schaffer,et al.  Characterization of integrin engagement during defined human embryonic stem cell culture , 2010, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[29]  Y. Okada,et al.  MT-MMP, the cell surface activator of proMMP-2 (pro-gelatinase A), is expressed with its substrate in mouse tissue during embryogenesis. , 1996, Journal of cell science.

[30]  K. Sekiguchi,et al.  Ligand-binding specificities of laminin-binding integrins: A comprehensive survey of laminin–integrin interactions using recombinant α3β1, α6β1, α7β1 and α6β4 integrins , 2006 .

[31]  J. D’Armiento,et al.  Matrix metalloproteinases in development and disease. , 2006, Birth defects research. Part C, Embryo today : reviews.

[32]  T. Ching,et al.  Distinct ligand binding sites in integrin α3β1 regulate matrix adhesion and cell–cell contact , 2003, The Journal of cell biology.

[33]  Bonnie F. Sloane,et al.  Degradation of laminin by human tumor cathepsin B , 1989, Clinical & Experimental Metastasis.

[34]  P. Yurchenco,et al.  Role of Laminin Terminal Globular Domains in Basement Membrane Assembly* , 2007, Journal of Biological Chemistry.

[35]  K. Chien,et al.  Long-term self-renewal of human pluripotent stem cells on human recombinant laminin-511 , 2010, Nature Biotechnology.

[36]  R. Fässler,et al.  The role of laminin in embryonic cell polarization and tissue organization. , 2003, Developmental cell.

[37]  S. Carbonetto,et al.  Mapping of Network-forming, Heparin-binding, and 11 Integrin-recognition Sites within the -Chain Short Arm of Laminin-1 (*) , 1995, The Journal of Biological Chemistry.

[38]  P. simon-Assmann.  The laminin family , 2013, Cell adhesion & migration.

[39]  C. Gilles,et al.  Epithelial-mesenchymal transition process in human embryonic stem cells cultured in feeder-free conditions. , 2007, Molecular human reproduction.

[40]  Raghu Kalluri,et al.  The epithelial–mesenchymal transition: new insights in signaling, development, and disease , 2006, The Journal of cell biology.

[41]  Z. Werb,et al.  Regulation of matrix biology by matrix metalloproteinases. , 2004, Current opinion in cell biology.

[42]  P. Stern,et al.  Epithelial-mesenchymal transition events during human embryonic stem cell differentiation. , 2007, Cancer research.

[43]  Y. Shintani,et al.  Cadherin switching , 2008, Journal of Cell Science.

[44]  H. Chapman,et al.  Integrin α3β1–dependent β-catenin phosphorylation links epithelial Smad signaling to cell contacts , 2009, The Journal of cell biology.

[45]  R. Mecham,et al.  Neutrophil Elastase Cleaves Laminin-332 (Laminin-5) Generating Peptides That Are Chemotactic for Neutrophils* , 2008, Journal of Biological Chemistry.

[46]  C. ffrench-Constant,et al.  Laminin enhances the growth of human neural stem cells in defined culture media , 2008, BMC Neuroscience.

[47]  G. Giannelli,et al.  Induction of cell migration by matrix metalloprotease-2 cleavage of laminin-5. , 1997, Science.

[48]  J. Kreidberg,et al.  (Alpha)3(beta)1 integrin regulates epithelial cytoskeletal organization. , 1999, Journal of cell science.

[49]  Trent P Munro,et al.  Stem cell integrins: implications for ex-vivo culture and cellular therapies. , 2011, Stem cell research.

[50]  E. Hohenester,et al.  Laminins in basement membrane assembly , 2013, Cell adhesion & migration.

[51]  G. Giannelli,et al.  Role of Cell Surface Metalloprotease Mt1-Mmp in Epithelial Cell Migration over Laminin-5 , 2000, The Journal of cell biology.

[52]  Ashley C. Brown,et al.  Guiding epithelial cell phenotypes with engineered integrin-specific recombinant fibronectin fragments. , 2011, Tissue engineering. Part A.

[53]  S. Shapiro,et al.  Matrix metalloproteinase degradation of extracellular matrix: biological consequences. , 1998, Current opinion in cell biology.

[54]  J. Miner,et al.  Laminin functions in tissue morphogenesis. , 2004, Annual review of cell and developmental biology.

[55]  W. Tourtellotte,et al.  EMMPRIN: A Novel Regulator of Leukocyte Transmigration into the CNS in Multiple Sclerosis and Experimental Autoimmune Encephalomyelitis , 2011, The Journal of Neuroscience.

[56]  P. Yurchenco,et al.  Self-assembly and calcium-binding sites in laminin. A three-arm interaction model. , 1993, The Journal of biological chemistry.