COMP-Ang1: a designed angiopoietin-1 variant with nonleaky angiogenic activity.

Angiopoietin-1 (Ang1) has potential therapeutic applications in inducing angiogenesis, enhancing endothelial cell survival, and preventing vascular leakage. However, production of Ang1 is hindered by aggregation and insolubility resulting from disulfide-linked higher-order structures. Here, by replacing the N-terminal portion of Ang1 with the short coiled-coil domain of cartilage oligomeric matrix protein (COMP), we have generated a soluble, stable, and potent Ang1 variant, COMP-Ang1. This variant is more potent than native Ang1 in phosphorylating the tyrosine kinase with Ig and epidermal growth factor homology domain 2 (Tie2) receptor and Akt in primary cultured endothelial cells, enhancing angiogenesis in vitro and increasing adult angiogenesis in vivo. Thus, COMP-Ang1 is an effective alternative to native Ang1 for therapeutic angiogenesis in vivo.

[1]  J. York,et al.  Tyrosine 1101 of Tie2 Is the Major Site of Association of p85 and Is Required for Activation of Phosphatidylinositol 3-Kinase and Akt , 1998, Molecular and Cellular Biology.

[2]  V. Malashkevich,et al.  The Crystal Structure of a Five-Stranded Coiled Coil in COMP: A Prototype Ion Channel? , 1996, Science.

[3]  N. Glazer,et al.  Angiopoietin-1 protects the adult vasculature against plasma leakage , 2000, Nature Medicine.

[4]  R. Auerbach,et al.  Angiogenesis in the mouse cornea. , 1979, Science.

[5]  Pamela F. Jones,et al.  Isolation of Angiopoietin-1, a Ligand for the TIE2 Receptor, by Secretion-Trap Expression Cloning , 1996, Cell.

[6]  J. Isner,et al.  Tie2 receptor ligands, angiopoietin-1 and angiopoietin-2, modulate VEGF-induced postnatal neovascularization. , 1998, Circulation research.

[7]  Thomas N. Sato,et al.  Increased vascularization in mice overexpressing angiopoietin-1. , 1998, Science.

[8]  J. Kim,et al.  Angiopoietin-1 regulates endothelial cell survival through the phosphatidylinositol 3'-Kinase/Akt signal transduction pathway. , 2000, Circulation research.

[9]  T. Sato,et al.  Tie-1 and tie-2 define another class of putative receptor tyrosine kinase genes expressed in early embryonic vascular system. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Chae,et al.  Coadministration of Angiopoietin-1 and Vascular Endothelial Growth Factor Enhances Collateral Vascularization , 2000, Arteriosclerosis, thrombosis, and vascular biology.

[11]  J. Isner,et al.  Direct intramuscular injection of plasmid DNA encoding angiopoietin-1 but not angiopoietin-2 augments revascularization in the rabbit ischemic hindlimb. , 1998, Circulation.

[12]  P. S. Kim,et al.  X-ray structure of the GCN4 leucine zipper, a two-stranded, parallel coiled coil. , 1991, Science.

[13]  Y. Pak,et al.  The angiopoietin-tie2 system in coronary artery endothelium prevents oxidized low-density lipoprotein-induced apoptosis. , 2001, Cardiovascular research.

[14]  U. Aebi,et al.  Preparation of single molecules and supramolecular complexes for high-resolution metal shadowing. , 1983, Journal of ultrastructure research.

[15]  Thomas N. Sato,et al.  Leakage-resistant blood vessels in mice transgenically overexpressing angiopoietin-1. , 1999, Science.

[16]  J. Isner,et al.  Angiogenesis and vasculogenesis as therapeutic strategies for postnatal neovascularization. , 1999, The Journal of clinical investigation.

[17]  William Lee,et al.  Angiopoietin-1 and -2 Coiled Coil Domains Mediate Distinct Homo-oligomerization Patterns, but Fibrinogen-like Domains Mediate Ligand Activity* , 1999, The Journal of Biological Chemistry.

[18]  G. Koh,et al.  Angiopoietin-1 inhibits irradiation- and mannitol-induced apoptosis in endothelial cells. , 2000, Circulation.

[19]  Ernest Fraenkel,et al.  Comparison of X-ray and NMR structures for the Antennapedia homeodomain–DNA complex , 1998, Nature Structural &Molecular Biology.

[20]  Thomas N. Sato,et al.  Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. , 1997, Science.

[21]  P. Carmeliet.  Mechanisms of angiogenesis and arteriogenesis , 2000, Nature Medicine.

[22]  Andrei T. Alexandrescu,et al.  NMR structure of a parallel homotrimeric coiled coil , 1998, Nature Structural &Molecular Biology.

[23]  G. Koh,et al.  Angiopoietin-1 induces endothelial cell sprouting through the activation of focal adhesion kinase and plasmin secretion. , 2000, Circulation research.

[24]  K. Alitalo,et al.  Identification of Tek/Tie2 Binding Partners , 1999, The Journal of Biological Chemistry.

[25]  G. Koh,et al.  Angiopoietin-1 Reduces VEGF-Stimulated Leukocyte Adhesion to Endothelial Cells by Reducing ICAM-1, VCAM-1, and E-Selectin Expression , 2001, Circulation research.

[26]  Pamela F. Jones,et al.  Requisite Role of Angiopoietin-1, a Ligand for the TIE2 Receptor, during Embryonic Angiogenesis , 1996, Cell.

[27]  Stanley J. Wiegand,et al.  Vascular-specific growth factors and blood vessel formation , 2000, Nature.

[28]  K. Alitalo,et al.  Tie receptors: new modulators of angiogenic and lymphangiogenic responses , 2001, Nature Reviews Molecular Cell Biology.

[29]  T. Noda,et al.  A Role for Hematopoietic Stem Cells in Promoting Angiogenesis , 2000, Cell.

[30]  Czeslaw Radziejewski,et al.  Moving across membranes , 2003, Nature Structural Biology.