VEGF-TRAPR1R2 suppresses choroidal neovascularization and VEGF-induced breakdown of the blood–retinal barrier†
Yoshitsugu Saishin
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Search for more papers by this authorYumiko Saishin
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Search for more papers by this authorKyoichi Takahashi
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Search for more papers by this authorRaquel Lima e Silva
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Search for more papers by this authorDonna Hylton
Regeneron Pharmaceuticals, Tarrytown, New York, New York
Search for more papers by this authorJohn S. Rudge
Regeneron Pharmaceuticals, Tarrytown, New York, New York
Search for more papers by this authorStanley J. Wiegand
Regeneron Pharmaceuticals, Tarrytown, New York, New York
Search for more papers by this authorCorresponding Author
Peter A. Campochiaro
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Maumenee 719, The Johns Hopkins University School of Medicine, 600 N. Wolfe Street, Baltimore, MD 21287-9277.Search for more papers by this authorYoshitsugu Saishin
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Search for more papers by this authorYumiko Saishin
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Search for more papers by this authorKyoichi Takahashi
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Search for more papers by this authorRaquel Lima e Silva
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Search for more papers by this authorDonna Hylton
Regeneron Pharmaceuticals, Tarrytown, New York, New York
Search for more papers by this authorJohn S. Rudge
Regeneron Pharmaceuticals, Tarrytown, New York, New York
Search for more papers by this authorStanley J. Wiegand
Regeneron Pharmaceuticals, Tarrytown, New York, New York
Search for more papers by this authorCorresponding Author
Peter A. Campochiaro
The Departments of Ophthalmology and Neuroscience, The Johns Hopkins University School of Medicine, Maumenee, Baltimore, Maryland
Maumenee 719, The Johns Hopkins University School of Medicine, 600 N. Wolfe Street, Baltimore, MD 21287-9277.Search for more papers by this authorPAC is the George S. and Dolores Dore Eccles Professor of Ophthalmology and Neuroscience.
Abstract
Vascular endothelial growth factor (VEGF) plays a central role in the development of retinal neovascularization and diabetic macular edema. There is also evidence suggesting that VEGF is an important stimulator for choroidal neovascularization. In this study, we investigated the effect of a specific inhibitor of VEGF, VEGF-TRAPR1R2, in models for these disease processes. VEGF-TRAPR1R2 is a fusion protein, which combines ligand binding elements taken from the extracellular domains of VEGF receptors 1 and 2 fused to the Fc portion of IgG1. Subcutaneous injections or a single intravitreous injection of VEGF-TRAPR1R2 strongly suppressed choroidal neovascularization in mice with laser-induced rupture of Bruch's membrane. Subcutaneous injection of VEGF-TRAPR1R2 also significantly inhibited subretinal neovascularization in transgenic mice that express VEGF in photoreceptors. In two models of VEGF-induced breakdown of the blood–retinal barrier (BRB), one in which recombinant VEGF is injected into the vitreous cavity and one in which VEGF expression is induced in the retina in transgenic mice, VEGF-TRAPR1R2 significantly reduced breakdown of the BRB. These data confirm that VEGF is a critical stimulus for the development of choroidal neovascularization and indicate that VEGF-TRAPR1R2 may provide a new agent for consideration for treatment of patients with choroidal neovascularization and diabetic macular edema. © 2003 Wiley-Liss, Inc.
LITERATURE CITED
- Amin R, Pulkin JE, Frank RN. 1994. Growth factor localization in choroidal neovascular membranes of age-related macular degeneration. Invest Ophthalmol Vis Sci 35: 3178–3188.
- Bainbridge J, Mistry A, Alwis MD, Paleolog E, Baker A, Thrasher AJ, Ali RR. 2002. Inhibition of retinal neovascularization by gene transfer of soluble VEGF receptor sFlt-1. Gene Ther 9: 320–326.
- Bold G, Altmann K-H, Bruggen J, Frei J, Lang M, Manley PW, Traxler P, Wietfeld B, Buchdunger E, Cozens R, Ferrari S, Furet P, Hofmann F, Martiny-Baron G, Mestan J, Rosel J, Sills M, Stover D, Acemonglu F, Boss E, Emmengger R, Lasser L, Masso E, Roth R, Schlachter C, Vetterli W, Wyss D, Wood JM. 2000. New anilino-phthalazines as potent and orally well absorbed inhibitors of the VEGF receptor tyrosine kinases useful as antagonists of tumor driven angiogenesis. J Med Chem 43: 2310–2323.
- Derevjanik NL, Vinores SA, Xiao W-H, Mori K, Turon T, Hudish T, Dong S, Campochiaro PA. 2002. Quantitative assessment of the integrity of the blood–retinal barrier in mice. Invest Ophthalmol Vis Sci 43: 2462–2467.
- Drevs J, Hofmann I, Hugenschmidt H, Wittig C, Madjar H, Muller M, Wood J, Martiny-Baron G, Unger C, Marme D. 2000. Effects of PTK787/ZK 222584, a specific inhibitor of vascular endothelial growth factor receptor tyrosine kinases, on primary tumor, metastasis, vessel density, and blood flow in a murine renal cell carcinoma model. Cancer Res 60: 4819–4824.
- Edelman JL, Castro MR. 2000. Quantitative image analysis of laser-induced choroidal neovascularization in rat. Exp Eye Res 71: 523–533.
- EyeTech Study Group. 2002. Preclinical and phase 1A clinical evaluation of an anti-VEGF pegylated aptamer (EYE001) for the treatment of exudative age-related macular degeneration. Retina 22: 143–152.
- Fabbro D, Buchdunger E, Wood J, Mestan J, Hofmann F, Ferrari S, Mett H, O'Reilly T, Meyer T. 1999. CGP 41251 a protein kinase inhibitor with potential as an anti-cancer agent. Pharmacol Ther 82: 293–298.
- Folkman J. 1995. Angiogenesis in cancer, vascular, rheumatoid, and other diseases. Nat Med 1: 27–31.
- Forsythe JA, Jiang B-H, Iyer NV, Agani F, Leung SW, Koos RD, Semenza G. 1996. Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol Cell Biol 16: 4604–4613.
- Frank RN, Amin RH, Eliott D, Puklin JE, Abrams GW. 1996. Basic fibroblast growth factor and vascular endothelial growth factor are present in epiretinal and choroidal neovascular membranes. Am J Ophthalmol 122: 393–403.
- Gerber H-P, Condorelli F, Park J, Ferrara N. 1997. Differential transcriptional regulation of the two vascular endothelial growth factor receptor genes. Flt-1, but not Flk-1/KDR, is up-regulated by hypoxia. J Biol Chem 272: 23659–23667.
- Goldman CK, Kendall RL, Cabrera G, Soroceanu L, Heike Y, Gillespie GY, Siegal GP, Mao X, Bett AJ, Huckle WR, Thomas KA, Curiel DT. 1998. Paracrine expression of a native soluble vascular endothelial growth factor receptor inhibits tumor growth, metastasis, and mortality rate. Proc Natl Acad Sci USA 95: 8795–8800.
- Grunwald J, Hariprasad S, DuPont J, Maguire M, Fine S, Brucker A, Maguire A, Ho A. 1998. Foveolar choroidal blood flow in age-related macular degeneration. Invest Ophthalmol Vis Sci 39: 385–390.
- Guyer DR, Martin DM, Klein M, Haller J, Group The EyeTech Study Group. 2001. Anti-VEGF therapy in patients with exudative age-related macular degeneration. Invest Ophthalmol Vis Sci (Suppl) 42: S522.
- Honda M, Sakamoto T, Ishibashi T, Inomata H, Ueno H. 2000. Experimental subretinal neovascularization is inhibited by adenovirus-mediated soluble VEGF.flt-1 receptor gene transfection: A role of VEGF and possible treatment for SRN in age-related macular degeneration. Gene Ther 7: 978–985.
- Iyer NV, Kotch LE, Agani F, Leung SW, Laughner E, Wenger RH, Gassmann M, Gearhart JD, Lawler AM, Yu AY, Semenza GL. 1998. Cellular and developmental control of O2 homeostasis by hypoxia-inducible factor 1alpha. Genes Dev 12: 149–162.
- Joussen AM, Poulaki V, Qin W, Kirchhof B, Mitsiades N, Wiegand SJ, Rudge GD, Yancopoulos G, Adamis AP. 2002. Retinal vascular endothelial growth factor induces intercellular adhesion molecule-1 and endothelial nitric oxide syntase expression and initiates early diabetic retinal leukocyte adhesion in vivo. Am J Pathol 160: 501–509.
- Klein R, Klein BEK, Moss SE, Davis MD, DeMets DL. 1984. The Wisconsin Epidemiologic Study of Diabetic Retinopathy. II. Prevalence and risk of diabetic retinopathy when age at diagnosis is less than 30 years. Arch Ophthalmol 102: 520–526.
- Kong H-L, Hecht D, Song W, Kovesdi I, Hackett NR, Yayon A, Crystal RG. 1998. Regional suppression of tumor growth by in vivo transfer of a cDNA encoding a secreted form of the extracellular domain of the flt-1 vascular endothelial growth factor receptor. Hum Gene Ther 9: 823–833.
- Kryzstolik MG, Afshari MA, Adamis AP, Gaudreault J, Gragoudas ES, Michaud NM, Li W, Connolly E, O'Neill CA, Miller JW. 2002. Prevention of experimental choroidal neovascularization with intravitreal anti-vascular endothelial growth factor antibody fragment. Arch Ophthalmol 120: 338–346.
- Kvanta A, Algvere PV, Berglin L, Seregard S. 1996. Subfoveal fibrovascular membranes in age-related macular degeneration express vascular endothelial growth factor. Invest Ophthalmol Vis Sci 37: 1929–1934.
- Kwak N, Okamoto N, Wood JM, Campochiaro PA. 2000. VEGF is an important stimulator in a model of choroidal neovascularization. Invest Ophthalmol Vis Sci 41: 3158–3164.
- Lai C-M, Brankov M, Zaknich T, Lai YK-Y, Shen W-Y, Constable IJ, Kovesdi I, Rakoczy PE. 2001. Inhibition of angiogenesis by adenovirus-mediated sFlt-1 expression in a rat model of corneal neovascularization. Hum Gene Ther 12: 1299–1310.
- Lai YK, Shen WY, Brankov M, Lai CM, Constable IJ, Rakoczy PE. 2002. Potential long-term inhibition of ocular neovascularization by recombinant adeno-associated virus-mediated secretion gene therapy. Gene Ther 9: 804–813.
- Lopez PF, Sippy BD, Lambert HM, Thach AB, Hinton DR. 1996. Transdifferentiated retinal pigment epithelial cells are immunoreactive for vascular endothelial growth factor in surgically excised age-related macular degeneration-related choroidal neovascular membranes. Invest Ophthalmol Vis Sci 37: 855–868.
- The Macular Photocoagulation Study. 1991. Argon laser photocoagulation for neovascular maculopathy: Five year results from randomized clinical trials. Arch Ophthalmol 109: 1109–1114.
- Mahasreshti PJ, Navarro JG, Kataram M, Wang MH, Carey D, Siegal GP, Barnes MN, Nettelbeck DM, Alvarez RD, Hemminki A, Curiel DT. 2001. Adenovirus-mediated soluble FLT-1 gene therapy for ovarian carcinoma. Clin Canc Res 7: 2057–2066.
- Mori K, Duh E, Gehlbach P, Ando A, Takahashi K, Pearlman J, Mori K, Yang HS, Zack DJ, Ettyreddy D, Brough DE, Wei LL, Campochiaro PA. 2001. Pigment epithelium-derived factor inhibits retinal and choroidal neovascularization. J Cell Physiol 188: 253–263.
- Ogata N, Matsushima M, Takada Y, Tobe T, Takahashi K, Yi X, Yamamoto C, Yamada H, Uyama M. 1996. Expression of basic fibroblast growth factor mRNA in developing choroidal neovascularization. Curr Eye Res 15: 1008–1018.
- Ohno-Matsui K, Hirose A, Yamamoto S, Saikia J, Okamoto N, Gehlbach P, Duh EJ, Hackett SF, Chang M, Bok D, Zack DJ, Campochiaro PA. 2002. Inducible expression of vascular endothelial growth factor in photoreceptors of adult mice causes severe proliferative retinopathy and retinal detachment. Am J Pathol 160: 711–719.
- Okamoto N, Tobe T, Hackett SF, Ozaki H, Vinores MA, LaRochelle W, Zack DJ, Campochiaro PA. 1997. Transgenic mice with increased expression of vascular endothelial growth factor in the retina: A new model of intraretinal and subretinal neovascularization. Am J Pathol 151: 281–291.
- Ozaki H, Hayashi H, Vinores SA, Moromizato Y, Campochiaro PA, Oshima K. 1997. Intravitreal sustained release of VEGF causes retinal neovascularization in rabbits and breakdown of the blood–retinal barrier in rabbits and primates. Exp Eye Res 64: 505–517.
- Ozaki H, Seo M-S, Ozaki K, Yamada H, Yamada E, Hofmann F, Wood J, Campochiaro PA. 2000. Blockade of vascular endothelial cell growth factor receptor signaling is sufficient to completely prevent retinal neovascularization. Am J Pathol 156: 679–707.
- Poulaki V, Qin W, Joussen AM, Hurlbit P, Wiegand SJ, Rudge J, Yancopoulos G, Adamis AP. 2002. Acute insulin therapy exacerbates diabetic blood–retinal barrier breakdown via HIF-1 alpha and vascular endothelial growth factor. J Clin Invest 109: 805–815.
- Qaum T, Xu Q, Joussen AM, Clemens MW, Qin W, Miyamoto K, Hassessian H, Wiegand SJ, Rudge J, Yancoupoulos GD, Adamis AP. 2001. VEGF-initiated blood–retinal barrier breakdown in early diabetes. Invest Ophthalmol Vis Sci 42: 2408–2413.
- Ross RD, Barofsky JM, Cohen G, Baber W, Palao Sh, Gitter K. 1998. Presumed macular choroidal watershed vascular filling, choroidal neovascularization, and systemic vascular disease in patients with age-related macular degeneration. Am J Ophthalmol 125: 71–80.
- Schwartz SD, Blumenkranz M, Rosenfeld PJ, Miller JW, Haller J, Fish G, Lobes L, Singerman L, Green WL, Reimann J. 2001. Safety of rhuFab V2, an anti-VEGF antibody fragment, as a single intravitreal injection in subjects with neovascular age-related macular degeneration. Invest Ophthalmol Vis Sci (Suppl) 42: S522.
- Seo M-S, Kwak N, Ozaki H, Yamada H, Okamoto N, Fabbro D, Hofmann F, Wood JM, Campochiaro PA. 1999. Dramatic inhibition of retinal and choroidal neovascularization by oral administration of a kinase inhibitor. Am J Pathol 154: 1743–1753.
- Shiose S, Sakamoto T, Yoshikawa H, Hata Y, Kawano Y, Ishibashi T, Inomata H, Takayama K, Ueno H. 2000. Gene transfer of a soluble receptor of VEGF inhibits the growth of experimental eyelid malignant melanoma. Invest Ophthalmol Vis Sci 41: 2395–2403.
- Takayama K, Ueno H, Nakanishi Y, Sakamoto T, Inoue K, Shimizu K, Oohashi H, Hai N. 2000. Suppression of tumor angiogenesis and growth by gene transfer of a soluble form of vascular endothelial growth factor receptor into a remote organ. Canc Res 60: 2168–2177.
- Tobe T, Okamoto N, Vinores MA, Derevjanik NL, Vinores SA, Zack DJ, Campochiaro PA. 1998a. Evolution of neovascularization in mice with overexpression of vascular endothelial growth factor in photoreceptors. Invest Ophthalmol Vis Sci 39: 180–188.
- Tobe T, Ortega S, Luna L, Ozaki H, Okamoto N, Derevjanik NL, Vinores SA, Basilico C, Campochiaro PA. 1998b. Targeted disruption of the FGF2 gene does not prevent choroidal neovascularization in a murine model. Am J Pathol 153: 1641–1646.
- Wong AK, Alfert M, Castrillon DH, Shen Q, Holash J, Yancopoulos GD, Chin L. 2001. Excessive tumor-elaborated VEGF and its neutralization define a lethal paraneoplastic syndrome. Proc Natl Acad Sci USA 98: 7481–7486.
- Wood JM, Bold G, Stover D, Hofmann F, Buchduneger E, Mestan J, Towbin H, Schnell C, Mett H, O'Reilly T, Cozens R, Rosel J, Siemeister G, Menrad A, Schirner M, Thierauch K-H, Schneider MR, Martiny-Baron G, Drevs J, Marme D. 2000. PTK787/ZK 222584, a novel and potent inhibitor of vascular endothelial growth factor receptor tyrosine kinases, impairs vascular endothelial growth factor-induced responses and tumor growth after oral administration. Cancer Res 60: 2178–2189.
- Wulff C, Wilson H, Wiegand SJ, Rudge JS, Fraser HM. 2002. Prevention of thecal angiogenesis, antral follicular growth and ovulation in the primate with vascular endothelial growth factor trapR1R2. Endocrinology 143: 2797–2807.
- Yi X, Ogata N, Komada M, Yamamoto C, Takahashi K, Omori K, Uyama M. 1997. Vascular endothelial growth factor expression in choroidal neovascularization in rats. Lab Invest 235: 313–319.
