Abstract
We evaluated the efficacy of equine infectious anaemia virus (EIAV)-based lentiviral vectors encoding endostatin (EIAV.endostatin) or angiostatin (EIAV.angiostatin) in inhibiting angiogenesis and vascular hyperpermeability in the laser-induced model of choroidal neovascularisation (CNV). Equine infectious anaemia virus.endostatin, EIAV.angiostatin or control (EIAV.null) vectors were administered into the subretinal space of C57Bl/6J mice. Two weeks after laser injury CNV areas and the degree of vascular hyperpermeability were measured by image analysis of in vivo fluorescein angiograms. Compared with EIAV.null-injected eyes, EIAV.endostatin resulted in a 59.5% (P<0.001) reduction in CNV area and a reduction in hyperpermeability of 25.6% (P<0.05). Equine infectious anaemia virus.angiostatin resulted in a 50.0% (P<0.05) reduction in CNV area and a 23.9% (P<0.05) reduction in hyperpermeability. Equine infectious anaemia virus.endostatin, but not EIAV.angiostatin significantly augmented the frequency of apoptosis within the induced CNV as compared with injected controls. TdT-dUTP terminal nick end labeling analysis 5 weeks post-injection, and histological and retinal flatmount analysis 12 months post-injection revealed no evidence of vector- or transgene expression-related deleterious effects on neurosensory retinal cells, or mature retinal vasculature in non-lasered eyes. Highly expressing EIAV-based vectors encoding endostatin or angiostatin effectively control angiogenesis and hyperpermeability in experimental CNV without long-term deleterious effects, supporting the use of such a strategy in the management of patients with exudative age-related macular degeneration.
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References
Macular Photocoagulation Study Group. Argon laser photocoagulation for neovascular maculopathy. Five-year results from randomized clinical trials. Arch Ophthalmol 1991; 109: 1109–1114.
Ferris III FL, Fine SL, Hyman L . Age-related macular degeneration and blindness due to neovascular maculopathy. Arch Ophthalmol 1984; 102: 1640–1642.
Guidelines for using verteporfin (Visudyne) in photodynamic therapy for choroidal neovascularization due to age-related macular degeneration and other causes: update. Retina 2005; 25: 119–134.
Lopez PF, Lambert HM, Grossniklaus HE, Sternberg Jr P . Well-defined subfoveal choroidal neovascular membranes in age-related macular degeneration. Ophthalmology 1993; 100: 415–422.
Moisseiev J, Alhalel A, Masuri R, Treister G . The impact of the macular photocoagulation study results on the treatment of exudative age-related macular degeneration. Arch Ophthalmol 1995; 113: 185–189.
Gragoudas ES, Adamis AP, Cunningham Jr ET, Feinsod M, Guyer DR . Pegaptanib for neovascular age-related macular degeneration. N Engl J Med 2004; 351: 2805–2816.
Husain D, Kim I, Gauthier D, Lane AM, Tsilimbaris MK, Ezra E et al. Safety and efficacy of intravitreal injection of ranibizumab in combination with verteporfin PDT on experimental choroidal neovascularization in the monkey. Arch Ophthalmol 2005; 123: 509–516.
Chang TS, Tonnu IQ, Globe DR, Fine J . Longitudinal changes in self-reported visual functioning in AMD patients in a randomized controlled Phase I/II trial of lucentisTM (ranizumab; rHuFAB v2). Invest Ophthalmol Visual Sci 2004; 45: E-abstract 3098.
Heier JS, Rosenfeld PJ, Antoszyk AN, Hantsbarger G, Kim R, Shams N . Long-term experience with lucentisTM (Ranibizumab) in patients with neovascular age-related macular degeneration (AMD). Invest Ophthalmol Visual Sci 2005; 46: E-abstract 1393.
Gaudreault J, Fei D, Rusit J, Suboc P, Shiu V . Preclinical pharmacokinetics of ranibizumab (rhuFabV2) after a single intravitreal administration. Invest Ophthalmol Vis Sci 2005; 46: 726–733.
Capone A, Macugen AMD Study Group. Intravitreous pegaptanib sodium (MacugenTM) in patients with age-related macular degeneration (AMD): safety and pharmacokinetics. Invest Ophthalmol Visual Sci 2005; 46: E-abstract 2362.
Bainbridge JW, Stephens C, Parsley K, Demaison C, Halfyard A, Thrasher AJ et al. In vivo gene transfer to the mouse eye using an HIV-based lentiviral vector; efficient long-term transduction of corneal endothelium and retinal pigment epithelium. Gene Therapy 2001; 8: 1665–1668.
Duisit G, Conrath H, Saleun S, Folliot S, Provost N, Cosset FL et al. Five recombinant simian immunodeficiency virus pseudotypes lead to exclusive transduction of retinal pigmented epithelium in rat. Mol Ther 2002; 6: 446–454.
Balaggan KS, Binley K, Esapa M, Iqball S, Askham Z, Kan O et al. Stable and efficient intraocular gene transfer using pseudotyped EIAV lentiviral vectors. J Gene Med 2006; 8: 275–285.
Issel CJ, Coggins L . Equine infectious anemia: current knowledge. J Am Vet Med Assoc 1979; 174: 727–733.
Sellon DC, Fuller FJ, McGuire TC . The immunopathogenesis of equine infectious anemia virus. Virus Res 1994; 32: 111–138.
O'Reilly MS, Holmgren L, Shing Y, Chen C, Rosenthal RA, Moses M et al. Angiostatin: a novel angiogenesis inhibitor that mediates the suppression of metastases by a Lewis lung carcinoma. Cell 1994; 79: 315–328.
O'Reilly MS, Boehm T, Shing Y, Fukai N, Vasios G, Lane WS et al. Endostatin: an endogenous inhibitor of angiogenesis and tumor growth. Cell 1997; 88: 277–285.
Dhanabal M, Ramchandran R, Waterman MJ, Lu H, Knebelmann B, Segal M et al. Endostatin induces endothelial cell apoptosis. J Biol Chem 1999; 274: 11721–11726.
Karumanchi SA, Jha V, Ramchandran R, Karihaloo A, Tsiokas L, Chan B et al. Cell surface glypicans are low-affinity endostatin receptors. Mol Cell 2001; 7: 811–822.
Rehn M, Veikkola T, Kukk-Valdre E, Nakamura H, Ilmonen M, Lombardo C et al. Interaction of endostatin with integrins implicated in angiogenesis. Proc Natl Acad Sci USA 2001; 98: 1024–1029.
Dixelius J, Larsson H, Sasaki T, Holmqvist K, Lu L, Engstrom A et al. Endostatin-induced tyrosine kinase signaling through the Shb adaptor protein regulates endothelial cell apoptosis. Blood 2000; 95: 3403–3411.
Zhang M, Yang Y, Yan M, Zhang J . Downregulation of vascular endothelial growth factor and integrinbeta(3) by endostatin in a mouse model of retinal neovascularization. Exp Eye Res 2005; 82: 74–80.
Hajitou A, Grignet C, Devy L, Berndt S, Blacher S, Deroanne CF et al. The antitumoral effect of endostatin and angiostatin is associated with a down-regulation of vascular endothelial growth factor expression in tumor cells. FASEB J 2002; 16: 1802–1804.
Kim YM, Hwang S, Kim YM, Pyun BJ, Kim TY, Lee ST et al. Endostatin blocks vascular endothelial growth factor-mediated signaling via direct interaction with KDR/Flk-1. J Biol Chem 2002; 277: 27872–27879.
Takahashi K, Saishin Y, Saishin Y, Silva RL, Oshima Y, Oshima S et al. Intraocular expression of endostatin reduces VEGF-induced retinal vascular permeability, neovascularization, and retinal detachment. FASEB J 2003; 17: 896–898.
Hari D, Beckett MA, Sukhatme VP, Dhanabal M, Nodzenski E, Lu H et al. Angiostatin induces mitotic cell death of proliferating endothelial cells. Mol Cell Biol Res Commun 2000; 3: 277–282.
Luo J, Lin J, Paranya G, Bischoff J . Angiostatin upregulates E-selectin in proliferating endothelial cells. Biochem Biophys Res Commun 1998; 245: 906–911.
Lucas R, Holmgren L, Garcia I, Jimenez B, Mandriota SJ, Borlat F et al. Multiple forms of angiostatin induce apoptosis in endothelial cells. Blood 1998; 92: 4730–4741.
Claesson-Welsh L, Welsh M, Ito N, Anand-Apte B, Soker S, Zetter B et al. Angiostatin induces endothelial cell apoptosis and activation of focal adhesion kinase independently of the integrin-binding motif RGD. Proc Natl Acad Sci USA 1998; 95: 5579–5583.
Moser TL, Stack MS, Asplin I, Enghild JJ, Hojrup P, Everitt L et al. Angiostatin binds ATP synthase on the surface of human endothelial cells. Proc Natl Acad Sci USA 1999; 96: 2811–2816.
Stack MS, Gately S, Bafetti LM, Enghild JJ, Soff GA . Angiostatin inhibits endothelial and melanoma cellular invasion by blocking matrix-enhanced plasminogen activation. Biochem J 1999; 340 (Part 1): 77–84.
Gupta N, Nodzenski E, Khodarev NN, Yu J, Khorasani L, Beckett MA et al. Angiostatin effects on endothelial cells mediated by ceramide and RhoA. EMBO Rep 2001; 2: 536–540.
Liu J, Razani B, Tang S, Terman BI, Ware JA, Lisanti MP . Angiogenesis activators and inhibitors differentially regulate caveolin-1 expression and caveolae formation in vascular endothelial cells. Angiogenesis inhibitors block vascular endothelial growth factor-induced down-regulation of caveolin-1. J Biol Chem 1999; 274: 15781–15785.
Redlitz A, Daum G, Sage EH . Angiostatin diminishes activation of the mitogen-activated protein kinases ERK-1 and ERK-2 in human dermal microvascular endothelial cells. J Vasc Res 1999; 36: 28–34.
Sima J, Zhang SX, Shao C, Fant J, Ma JX . The effect of angiostatin on vascular leakage and VEGF expression in rat retina. FEBS Lett 2004; 564: 19–23.
Mori K, Ando A, Gehlbach P, Nesbitt D, Takahashi K, Goldsteen D et al. Inhibition of choroidal neovascularization by intravenous injection of adenoviral vectors expressing secretable endostatin. Am J Pathol 2001; 159: 313–320.
Auricchio A, Behling KC, Maguire AM, O'Connor EM, Bennett J, Wilson JM et al. Inhibition of retinal neovascularization by intraocular viral-mediated delivery of anti-angiogenic agents. Mol Ther 2002; 6: 490–494.
Lai CC, Wu WC, Chen SL, Xiao X, Tsai TC, Huan SJ et al. Suppression of choroidal neovascularization by adeno-associated virus vector expressing angiostatin. Invest Ophthalmol Vis Sci 2001; 42: 2401–2407.
Gruter O, Kostic C, Crippa SV, Perez M-TR, Zografos L, Schorderet DF et al. Lentiviral vector-mediated gene transfer in adult mouse photoreceptors is impaired by the presence of a physical barrier. Gene Ther 2005; 12: 942–947.
Yao XY, Hageman GS, Marmor MF . Retinal adhesiveness is weakened by enzymatic modification of the interphotoreceptor matrix in vivo. Invest Ophthalmol Vis Sci 1990; 31: 2051–2058.
Mori K, Duh E, Gehlbach P, Ando A, Takahashi K, Pearlman J et al. Pigment epithelium-derived factor inhibits retinal and choroidal neovascularization. J Cell Physiol 2001; 188: 253–263.
Mori K, Gehlbach P, Yamamoto S, Duh E, Zack DJ, Li Q et al. AAV-mediated gene transfer of pigment epithelium-derived factor inhibits choroidal neovascularization. Invest Ophthalmol Vis Sci 2002; 43: 1994–2000.
Kwak N, Okamoto N, Wood JM, Campochiaro PA . VEGF is major stimulator in model of choroidal neovascularization. Invest Ophthalmol Vis Sci 2000; 41: 3158–3164.
Shen WY, Yu MJ, Barry CJ, Constable IJ, Rakoczy PE . Expression of cell adhesion molecules and vascular endothelial growth factor in experimental choroidal neovascularisation in the rat. Br J Ophthalmol 1998; 82: 1063–1071.
Brankin B, Campbell M, Canning P, Gardiner TA, Stitt AW . Endostatin modulates VEGF-mediated barrier dysfunction in the retinal microvascular endothelium. Exp Eye Res 2005; 81: 22–31.
Dixelius J, Cross M, Matsumoto T, Sasaki T, Timpl R, Claesson-Welsh L . Endostatin regulates endothelial cell adhesion and cytoskeletal organization. Cancer Res 2002; 62: 1944–1947.
Russ PK, Davidson MK, Hoffman LH, Haselton FR . Partial characterization of the human retinal endothelial cell tight and adherens junction complexes. Invest Ophthalmol Vis Sci 1998; 39: 2479–2485.
Mazarakis ND, Azzouz M, Rohll JB, Ellard FM, Wilkes FJ, Olsen AL et al. Rabies virus glycoprotein pseudotyping of lentiviral vectors enables retrograde axonal transport and access to the nervous system after peripheral delivery. Hum Mol Genet 2001; 10: 2109–2121.
Mitrophanous K, Yoon S, Rohll J, Patil D, Wilkes F, Kim V et al. Stable gene transfer to the nervous system using a non-primate lentiviral vector. Gene Therapy 1999; 6: 1808–1818.
Kingsman SM, Mitrophanous K, Olsen JC . Potential oncogene activity of the woodchuck hepatitis post-transcriptional regulatory element (WPRE). Gene Therapy 2005; 12: 3–4.
Wong LF, Scott Ralph G, Walmsley LE, Bienemann AS, Parham S, Kingsman SM et al. Lentiviral-mediated delivery of Bcl-2 or GDNF protects against excitotoxicity in the rat hippocampus. Mol Ther 2005; 11: 89–95.
Hobson AH, Donovan M, Humphries MM, Tuohy G, Nally M, Carmody R et al. Apoptotic Photoreceptor Death in the Rhodopsin Knockout Mouse in the Presence and Absence of c-fos. Exp Eye Res 2000; 71: 247–254.
Acknowledgements
We thank Professor Sue Kingsman for reviewing this manuscript. We also acknowledge the Special Trustees of Moorfields Eye Hospital, London.
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Balaggan, K., Binley, K., Esapa, M. et al. EIAV vector-mediated delivery of endostatin or angiostatin inhibits angiogenesis and vascular hyperpermeability in experimental CNV. Gene Ther 13, 1153–1165 (2006). https://doi.org/10.1038/sj.gt.3302769
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DOI: https://doi.org/10.1038/sj.gt.3302769
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