Reduction of endogenous angiogenesis inhibitors in Bruch's membrane of the submacular region in eyes with age-related macular degeneration.

OBJECTIVES To determine the relative levels of 3 potent inhibitors of angiogenesis (endostatin, pigment epithelium-derived factor, and thrombospondin 1) in the retinal pigment epithelium-Bruch's membrane-choriocapillaris complex in the submacular region in aged control eyes and eyes with age-related macular degeneration (AMD). METHODS Immunohistochemical analysis with antibodies against endostatin, pigment epithelium-derived factor, and thrombospondin 1 was performed on the macular region of aged control donor eyes (n = 8; mean age, 79.8 years) and eyes with AMD (n = 12; mean age, 83.9 years). Three independent masked observers scored the reaction product (scored from 0-7). Mean scores from the control eyes and the eyes with AMD were analyzed using 1-way analysis of variance and unpaired t test. RESULTS In control eyes, strong immunoreactivity of all 3 inhibitors was observed in the retinal pigment epithelium-Bruch's membrane-choriocapillaris complex. Immunoreactivity for endostatin, pigment epithelium-derived factor, and thrombospondin 1 in Bruch's membrane was significantly lower in eyes with AMD compared with aged control eyes (analysis of variance, P = .003, P = .009, and P < .001, respectively). In the choriocapillaris, a significant reduction was observed in endostatin (analysis of variance, P = .02) and thrombospondin 1 (analysis of variance, P = .005) in eyes with AMD. CONCLUSIONS These findings suggest that endogenous angiogenesis inhibitors in the retinal pigment epithelium-Bruch's membrane-choriocapillaris complex may provide a biochemical barrier for choroidal neovascular invasion. CLINICAL RELEVANCE Decreased levels of angiogenic inhibitors at the retinal pigment epithelium-Bruch's membrane-choriocapillaris complex in eyes with AMD make Bruch's membrane vulnerable to choroidal neovascularization.

[1]  V. Gahtan,et al.  The role of thrombospondin-1 in human disease. , 2004, The Journal of surgical research.

[2]  Marco A Zarbin,et al.  Current concepts in the pathogenesis of age-related macular degeneration. , 2004, Archives of ophthalmology.

[3]  O. Volpert,et al.  Peptides derived from two separate domains of the matrix protein thrombospondin-1 have anti-angiogenic activity , 1993, The Journal of cell biology.

[4]  M. Yacoub,et al.  Antigenic heterogeneity of vascular endothelium. , 1992, The American journal of pathology.

[5]  C. Sorenson,et al.  Thrombospondin‐1–deficient mice exhibit increased vascular density during retinal vascular development and are less sensitive to hyperoxia‐mediated vessel obliteration , 2003, Developmental dynamics : an official publication of the American Association of Anatomists.

[6]  L. Liotta,et al.  Platelet thrombospondin modulates endothelial cell adhesion, motility, and growth: a potential angiogenesis regulatory factor , 1990, The Journal of cell biology.

[7]  P. Carmeliet,et al.  Angiogenesis in cancer and other diseases , 2000, Nature.

[8]  R. Timpl,et al.  Structure, function and tissue forms of the C‐terminal globular domain of collagen XVIII containing the angiogenesis inhibitor endostatin , 1998, The EMBO journal.

[9]  J. Lahav,et al.  Thrombospondin inhibits adhesion of endothelial cells. , 1988, Experimental cell research.

[10]  N. Bouck.  PEDF: anti-angiogenic guardian of ocular function. , 2002, Trends in molecular medicine.

[11]  F. Incardona,et al.  Thrombospondin-1 expression in RPE and choroidal neovascular membranes. , 2006, Yan ke xue bao = Eye science.

[12]  A. Ljubimov,et al.  Expression of protein kinase CK2 in astroglial cells of normal and neovascularized retina. , 2006, The American journal of pathology.

[13]  Richard F Spaide,et al.  CHOROIDAL NEOVASCULARIZATION IN AGE-RELATED MACULAR DEGENERATION—WHAT IS THE CAUSE? , 2003, Retina.

[14]  P. Jong,et al.  Immunohistochemical light and electron microscopy of basal laminar deposit , 2004, Graefe's Archive for Clinical and Experimental Ophthalmology.

[15]  J. Folkman.  Clinical Applications of Research on Angiogenesis , 1995 .

[16]  D. Lefer,et al.  Enhanced expression of intracellular adhesion molecule-1 and P-selectin in the diabetic human retina and choroid. , 1995, The American journal of pathology.

[17]  Peter Gehlbach,et al.  Pigment epithelium-derived factor suppresses ischemia-induced retinal neovascularization and VEGF-induced migration and growth. , 2002, Investigative ophthalmology & visual science.

[18]  J. Folkman.  Opinion: Angiogenesis: an organizing principle for drug discovery? , 2007, Nature Reviews Drug Discovery.

[19]  J. Tombran-Tink,et al.  PEDF: a pigment epithelium-derived factor with potent neuronal differentiative activity. , 1991, Experimental eye research.

[20]  H. Krutzsch,et al.  Modulation of endothelial cell proliferation, adhesion, and motility by recombinant heparin‐binding domain and synthetic peptides from the type I repeats of thrombospondin , 1993, Journal of cellular biochemistry.

[21]  G. Lutty,et al.  Heterogeneity in localization of isoforms of TGF-beta in human retina, vitreous, and choroid. , 1993, Investigative ophthalmology & visual science.

[22]  D. S. Mcleod,et al.  Pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF) in aged human choroid and eyes with age-related macular degeneration. , 2006, Experimental eye research.

[23]  N. Ferrara.  Vascular endothelial growth factor: molecular and biological aspects. , 1999, Current topics in microbiology and immunology.

[24]  P. Campochiaro,et al.  Clinical protocol. An open-label, phase I, single administration, dose-escalation study of ADGVPEDF.11D (ADPEDF) in neovascular age-related macular degeneration (AMD). , 2001, Human gene therapy.

[25]  J. Folkman.  Seminars in Medicine of the Beth Israel Hospital, Boston. Clinical applications of research on angiogenesis. , 1995, The New England journal of medicine.

[26]  J. Bennett,et al.  Inhibition of retinal neovascularization by intraocular viral-mediated delivery of anti-angiogenic agents. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[27]  I Que,et al.  Endostatin inhibits VEGF‐induced endothelial cell migration and tumor growth independently of zinc binding , 1999, The EMBO journal.

[28]  M. Pepper.  Role of the Matrix Metalloproteinase and Plasminogen Activator-Plasmin Systems in Angiogenesis , 2001, Arteriosclerosis, thrombosis, and vascular biology.

[29]  U. Felbor,et al.  Collagen XVIII/endostatin structure and functional role in angiogenesis. , 2000, Cell structure and function.

[30]  W R Green,et al.  Senile macular degeneration: a histopathologic study. , 1977, Transactions of the American Ophthalmological Society.

[31]  内田 博子,et al.  Production and accumulation of thrombospondin-1 in human retinal pigment epithelial cells , 2001 .

[32]  L. Cornelius,et al.  Thrombospondin-1, a natural inhibitor of angiogenesis, is present in vitreous and aqueous humor and is modulated by hyperglycemia. , 2000, Biochemical and biophysical research communications.

[33]  I. Grierson,et al.  Thrombospondin as a component of the extracellular matrix of epiretinal membranes: Comparisons with cellular fibronectin , 1992, Eye.

[34]  S. Becerra.  Focus on Molecules: Pigment epithelium-derived factor (PEDF). , 2006, Experimental eye research.

[35]  Joyce Tombran-Tink,et al.  Expression of pigment epithelium-derived factor in normal adult rat eye and experimental choroidal neovascularization. , 2002, Investigative ophthalmology & visual science.

[36]  Sonia H Yoo,et al.  Age-related macular degeneration: etiology, pathogenesis, and therapeutic strategies. , 2003, Survey of ophthalmology.

[37]  S. Campbell,et al.  Molecular mediators of angiogenesis in bladder cancer. , 1998, Cancer research.

[38]  M. Cross,et al.  Angiostatin and endostatin inhibit endothelial cell migration in response to FGF and VEGF without interfering with specific intracellular signal transduction pathways , 2003, FEBS letters.

[39]  B. Olsen,et al.  Localization of collagen XVIII and the endostatin portion of collagen XVIII in aged human control eyes and eyes with age-related macular degeneration. , 2004, Investigative ophthalmology & visual science.

[40]  J. Debus,et al.  Endostatin's antiangiogenic signaling network. , 2004, Molecular cell.

[41]  P. Campochiaro,et al.  Inhibition of choroidal neovascularization by intravenous injection of adenoviral vectors expressing secretable endostatin. , 2001, The American journal of pathology.

[42]  U. Felbor,et al.  Generation and degradation of human endostatin proteins by various proteinases , 2000, FEBS letters.

[43]  Tiansen Li,et al.  Collagen XVIII/endostatin is essential for vision and retinal pigment epithelial function , 2004, The EMBO journal.

[44]  G. Chader.  PEDF: Raising both hopes and questions in controlling angiogenesis , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[45]  J. Bennett,et al.  Gene therapy for ocular neovascularization: a cure in sight. , 2003, Current opinion in genetics & development.

[46]  William Arbuthnot Sir Lane,et al.  Endostatin: An Endogenous Inhibitor of Angiogenesis and Tumor Growth , 1997, Cell.

[47]  N. Bouck,et al.  Prevention of ischemia-induced retinopathy by the natural ocular antiangiogenic agent pigment epithelium-derived factor , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[48]  K. Talks,et al.  Current Status Of Antiangiogenic Factors , 2000, British journal of haematology.

[49]  P. Campochiaro,et al.  Regression of ocular neovascularization in response to increased expression of pigment epithelium-derived factor. , 2002, Investigative ophthalmology & visual science.

[50]  J. Ji.  Pigment epithelium derived factor:a potent inhibitor of angiogenesis , 2002 .

[51]  D. S. Mcleod,et al.  Impaired expression of thrombospondin-1 in eyes with age related macular degeneration , 2005, British Journal of Ophthalmology.