Molecular properties of fibrin-based matrices for promotion of angiogenesis in vitro.

The molecular properties of fibrin-based matrices, such as fibrillar structure and covalent modifications with adhesion domains, influence the angiogenic behavior of human umbilical vein endothelial cells (HUVECs) in vitro. The fibrillar structure of fibrin-based matrices was influenced by pH but not by covalent incorporation of exogenous adhesion domains. Native fibrin-based matrices polymerized at pH 10 formed organized and longitudinally oriented fibrin fibrils, which provided a good angiogenic substrate for endothelial cells. Furthermore, upon covalent incorporation of the model ligand L1Ig6, which binds to the integrin most prominently expressed on the surface of angiogenic endothelial cells, alpha(v)beta3, these matrices became angiogenesis-promoting when polymerized at physiological pH. The amount of incorporation of L1Ig6 into the matrices depended on the fibrinogen concentration on all three fibrin chains. Soluble forms of L1Ig6 diffused rapidly out of the matrix. Most important, L1Ig6-modified matrices were very specific in inducing the angiogenic phenotype of HUVECs, whereas control cells did not differentiate on these matrices. Our results indicate that artificial extracellular matrices can influence cell behavior in two ways. One way is based on the three-dimensional fibril structure of the matrix molecules themselves, and the other is due to providing specific binding sites for direct cell-matrix interactions that lead to the activation of second-messenger cascades and thus promoting angiogenic differentiation.

[1]  M. Ginsberg,et al.  Breaking the Integrin Hinge , 1996, The Journal of Biological Chemistry.

[2]  J. Heino,et al.  Adhesion receptors and cell invasion: mechanisms of integrin-guided degradation of extracellular matrix , 2000, Cellular and Molecular Life Sciences CMLS.

[3]  M. Humphries,et al.  Integrin activation: the link between ligand binding and signal transduction. , 1996, Current opinion in cell biology.

[4]  W. D. de Jong,et al.  Substrate Requirements for Transglutaminases , 1995, The Journal of Biological Chemistry.

[5]  J. Folkman Angiogenesis in cancer, vascular, rheumatoid and other disease , 1995, Nature Medicine.

[6]  Richard O. Hynes,et al.  Integrins: Versatility, modulation, and signaling in cell adhesion , 1992, Cell.

[7]  T. Tamaki,et al.  Factor XIII‐mediated cross‐linking of NH2‐terminal peptide of α2‐plasmin inhibitor to fibrin , 1983 .

[8]  D. Teplow,et al.  Biosynthesis and membrane topography of the neural cell adhesion molecule L1. , 1985, The EMBO journal.

[9]  X. Yang,et al.  A potential role for the plasmin(ogen) system in the posttranslational cleavage of the neural cell adhesion molecule L1. , 1999, Journal of cell science.

[10]  R. Soldi,et al.  Role of αvβ3 integrin in the activation of vascular endothelial growth factor receptor‐2 , 1999, The EMBO journal.

[11]  Vance Lemmon,et al.  L1-mediated axon outgrowth occurs via a homophilic binding mechanism , 1989, Neuron.

[12]  S. Kumar,et al.  Hyaluronan stimulates tumor cell migration by modulating the fibrin fiber architecture. , 1999, Journal of cell science.

[13]  M. Humphries,et al.  The molecular basis and specificity of integrin-ligand interactions. , 1990, Journal of cell science.

[14]  S. Dedhar,et al.  Integrin cytoplasmic interactions and bidirectional transmembrane signalling. , 1996, Current opinion in cell biology.

[15]  Donald E. Ingber,et al.  How does extracellular matrix control capillary morphogenesis? , 1989, Cell.

[16]  C. Bucana,et al.  Regulation of distinct steps of angiogenesis by different angiogenic molecules. , 1998, International journal of oncology.

[17]  J. Weisel Fibrin assembly. Lateral aggregation and the role of the two pairs of fibrinopeptides. , 1986, Biophysical journal.

[18]  W. Risau,et al.  Mechanisms of angiogenesis , 1997, Nature.

[19]  S. Mahooti,et al.  Distinct signal transduction pathways are utilized during the tube formation and survival phases of in vitro angiogenesis. , 1998, Journal of cell science.

[20]  D. Sheppard,et al.  Plasmin-Sensitive Dibasic Sequences in the Third Fibronectin-like Domain of L1–Cell Adhesion Molecule (CAM) Facilitate Homomultimerization and Concomitant Integrin Recruitment , 2000, The Journal of cell biology.

[21]  Napoleone Ferrara,et al.  Clinical applications of angiogenic growth factors and their inhibitors , 1999, Nature Medicine.

[22]  C. Lagenaur,et al.  An L1-like molecule, the 8D9 antigen, is a potent substrate for neurite extension. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[23]  A. Horwitz,et al.  Integrin cytoplasmic domains: mediators of cytoskeletal linkages and extra- and intracellular initiated transmembrane signaling. , 1993, Current opinion in cell biology.

[24]  V. Nehls,et al.  The configuration of fibrin clots determines capillary morphogenesis and endothelial cell migration. , 1996, Microvascular research.

[25]  M. Schachner,et al.  Immunocytological and biochemical characterization of a new neuronal cell surface component (L1 antigen) which is involved in cell adhesion. , 1984, The EMBO journal.

[26]  M. Bourassa,et al.  Management of coronary artery disease: therapeutic options in patients with diabetes. , 2000, Journal of the American College of Cardiology.

[27]  S. Kenwrick,et al.  Neural cell adhesion molecule L1: relating disease to function , 1998, BioEssays : news and reviews in molecular, cellular and developmental biology.

[28]  R. Kammerer,et al.  Structural Analysis of the Sixth Immunoglobulin‐Like Domain of Mouse Neural Cell Adhesion Molecule L1 and Its Interactions with αvβ3, αIIbβ3, and α5β1 Integrins , 1998 .

[29]  J. Hubbell,et al.  Cross-linking exogenous bifunctional peptides into fibrin gels with factor XIIIa. , 1999, Bioconjugate chemistry.

[30]  W. Freed,et al.  Degradation fragments of L1 antigen enhance tyrosine hydroxylase-immunoreactive neurite outgrowth in mesencephalic cell culture , 1993, Brain Research.

[31]  S. Mandriota,et al.  Vascular endothelial growth factor-induced in vitro angiogenesis and plasminogen activator expression are dependent on endogenous basic fibroblast growth factor. , 1997, Journal of cell science.

[32]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[33]  J. Engel,et al.  Trimerization of Cell Adhesion Molecule L1 Mimics Clustered L1 Expression on the Cell Surface , 2000, Journal of neurochemistry.

[34]  D. Cheresh,et al.  Role of alpha v integrins during angiogenesis. , 2000, Cancer journal.

[35]  David A. Calderwood,et al.  Integrins and Actin Filaments: Reciprocal Regulation of Cell Adhesion and Signaling* , 2000, The Journal of Biological Chemistry.

[36]  L. Lorand,et al.  Influence of a natural and a synthetic inhibitor of factor XIIIa on fibrin clot rheology. , 1999, Biophysical journal.

[37]  M. Schachner,et al.  Biochemical Characterization of Different Molecular Forms of the Neural Cell Adhesion Molecule L1 , 1988, Journal of neurochemistry.

[38]  J. Becker,et al.  Human neural cell adhesion molecule L1 and rat homologue NILE are ligands for integrin alpha v beta 3 , 1996, The Journal of cell biology.

[39]  E. Engvall,et al.  Integrins and vascular extracellular matrix assembly. , 1997, The Journal of clinical investigation.

[40]  M. Iruela-Arispe,et al.  Modulation of extracellular matrix proteins by endothelial cells undergoing angiogenesis in vitro. , 1991, Arteriosclerosis and thrombosis : a journal of vascular biology.

[41]  M. Wilchek,et al.  [14] Protein biotinylation , 1990 .

[42]  D. Cheresh,et al.  Requirement of vascular integrin alpha v beta 3 for angiogenesis. , 1994, Science.

[43]  L. Greene,et al.  Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. , 1976, Proceedings of the National Academy of Sciences of the United States of America.

[44]  M. Hortsch Structural and Functional Evolution of the L1 Family: Are Four Adhesion Molecules Better Than One? , 2000, Molecular and Cellular Neuroscience.

[45]  W. Freed,et al.  Cell adhesion molecules (CAMs) in adrenal medulla in situ and in vitro: Enhancement of chromaffin cell L1 Ng-CAM expression by NGF , 1990, Experimental Neurology.

[46]  M. Schachner,et al.  L1/HNK‐1 Carbohydrate‐ and β1 Integrin‐Dependent Neural Cell Adhesion to Laminin‐1 , 1997, Journal of neurochemistry.

[47]  D. Cheresh,et al.  A Single Immunoglobulin-like Domain of the Human Neural Cell Adhesion Molecule L1 Supports Adhesion by Multiple Vascular and Platelet Integrins , 1997, The Journal of cell biology.

[48]  M. Schwartz,et al.  The extracellular matrix as a cell survival factor. , 1993, Molecular biology of the cell.

[49]  L. Orci,et al.  Angiogenesis: a paradigm for balanced extracellular proteolysis during cell migration and morphogenesis. , 1996, Enzyme & protein.