Adenovirus-mediated intravitreal delivery of Tum5 inhibits retinal neovascularization in a mouse model of oxygen-induced retinopathy_Chinese Journal of Ocular Fundus Diseases

Authors：

SunJing , YangWei , ZhangYan , 薄其玉 ,  张红

Tianjin Medical University Hospital, Tianjin Medical University Eye Institute, College of Optometry and Ophthalmology, Tianjin Medical University, Tianjin 300384, China;

Corresponding author：

张红, Email: tmuechong@sina.com

Keywords：

Retinal neovascularization/prevention & control; Gene transfer techniques; Transfection; Animal experimentation

DOI：

10.3760/cma.j.issn.1005-1015.2015.04.015

Video：

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Objective To observe the effect of adenovirus-mediated Tum5 (rAd-Tum5) inhibiting retinal neovascularization (RNV) of oxygen-induced retinopathy (OIR) mouse model. Methods The OIR model was induced in 96 C57BL/6J mice aged of 7 days according to the literature. These mice were divided randomly into control group, OIR group, OIR rAd-green fluorescent grotein (GFP) group and OIR rAd-Tum5 group, each group had 24 mice. The rAd-GFP and rAd-Tum5 were injected into the vitreous cavity of mice aged of 12 days in OIR rAd-GFP group and OIR rAd-Tum5 group, respectively. Meanwhile, OIR group and the control group received the injection of physiological saline solution of same volume. The relatively non-perfusion area was evaluated by fluorescence angiography, and the number of pre-retinal nucleus breaking through internal limiting membranes was observed by hematoxylin-eosin staining. The expression of vascular endothelial growth factor (VEGF) was estimated by immunofluorescent (IF) and Western blot. Results The retinal avascular areas of all groups were significantly different (F=61.224, P<0.01). The retinal avascular area of the rAd-Tum5 group was decreased significantly comparing with that in the OIR group and rAd-GFP group (P<0.01). However, there are no significant differences between the OIR group and rAd-GFP group (P=0.827). The number of pre-retinal nucleus breaking through ILM of all groups was significantly different (F=635.738, P<0.01), but no significantly difference was observed in OIR group and rAd-GFP group (P=0.261). Significant differences could also been seen between OIR rAd-Tum5 group and OIR group as well as OIR rAd-Tum5 group and OIR rAd-GFP group (P<0.01). The results of IF and Western blot indicated that expression of VEGF in the OIR group and rAd-GFP group was obviously up-regulated, compared with that in the control group. But the expression was declined in the rAd-Tum5 group compared with that in the OIR group and rAd-GFP group. Conclusion Tum-5 peptide can efficiently prevent RNV probably by down-regulating expression of VEGF.

Citation： SunJing, YangWei, ZhangYan. Adenovirus-mediated intravitreal delivery of Tum5 inhibits retinal neovascularization in a mouse model of oxygen-induced retinopathy. Chinese Journal of Ocular Fundus Diseases, 2015, 31(4): 371-376. doi: 10.3760/cma.j.issn.1005-1015.2015.04.015 Copy

1.	Ye HX, Yao Y, Jiang XJ, et al. Tumstatin transfected into human glioma cell line U251 represses tumor growth by inhibiting angiogenesis[J]. Chin Med J (Engl),2013,126(9):1720-1725.
2.	You Y, Xue X, Li M, et al. Inhibition effect of pcDNA-tum-5 on the growth of S180 tumor[J]. Cytotechnology,2008,56(2):97-104.
3.	Thevenard J, Ramont L, Devy J, et al. The YSNSG cyclopeptide derived from tumstatin inhibits tumor angiogenesis by down-regulating endothelial cell migration[J]. Int J Cancer,2010,126(5):1055-1066.
4.	黄敏丽,罗国容,陈维平,等. Tumstatin肽对视网膜微血管内皮细胞增生和迁移的影响[J]. 眼科新进展,2008,28(9):649-652.
5.	Esipov R, Beyrakhova K, Likhvantseva V, et al. Antiangiogenic and antivascular effects of a recombinant tumstatin-derived peptide in a corneal neovascularization model[J]. Biochimie,2012,94(6):1368-1375.
6.	Maeshima Y, Sudhakar A, Lively JC, et al. Tumstatin, an endothelial cell-specific inhibitor of protein synthesis[J]. Science,2002,295(5552):140-143.
7.	Maeshima Y, Colorado PC, Torre A,et al. Distinct antitumor properties of a type Ⅳ collagen domain derived from basement membrane[J]. J Biol Chem, 2000, 275(28):21340-21348.
8.	Ricci B, Lepore D, Iossa M, et al. Effect of light on oxygen-induced retinopathy in the rat model: light and OIR in the rat[J]. Doc Ophthalmol, 1990, 74(4):287-301.
9.	Smith LE, Wesolowski E, Mclellan A, et al. Oxygen-induced retinopathy in the mouse[J]. Invest Ophthalmol Vis Sci,1994,35(1):101-111.
10.	Li S, Li T, Luo Y, et al. Retro-orbital injection of FITC-dextran is an effective and economical method for observing mouse retinal vessels[J]. Mol Vis, 2011,17:3566-3573.
11.	Maeshima Y, Manfredi M, Reimer C, et al. Identification of the anti-angiogenic site within vascular basement membrane-derived Tumstatin[J]. J Biol Chem, 2001,276(18):15240-15248.
12.	Maeshima Y, Yerramalla UL, Dhanabal M,et al.Extracellular matrix-derived peptide binds to alpha vbeta 3 integrin and inhibits angiogenesis[J].J Biol Chem,2001,276(34):31959-31968.
13.	Wang L, Shi P, Xu Z, et al. Up-regulation of VEGF by retinoic acid during hyperoxia prevents retinal neovascularization and retinopathy[J]. Invest Ophthalmol Vis Sci,2014,55(7):4276-4287.
14.	Ameri H, Liu H, Liu R, et al. TWEAK/Fn14 pathway is a novel mediator of retinal neovascularization[J]. Invest Ophthalmol Vis Sci,2014,55(2):801-813.
15.	Li Z, He T, Du K, et al. Inhibition of oxygen-induced ischemic retinal neovascularization with adenoviral 15-lipoxygenase-1 gene transfer via up-regulation of PPAR-γ and Down-regulation of VEGFR-2 expression[J/OL]. PLoS ONE, 2014,9(1):85824[2014-01-21]. http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0085824.
16.	Liu G, Wu J, Tsai H, et al. Proopiomelanocortin gene delivery induces apoptosis in melanoma through NADPH oxidase 4-mediated ROS generation[J].Free Radic Biol Med, 2014,70:14-22.
17.	Maclaren RE, Groppe M, Barnard AR, et al. Retinal gene therapy in patients with choroideremia: initial findings from a phase 1/2 clinical trial[J]. Lancet, 2014,383(9923):1129-1137.
18.	王爱媛,高殿文,王桂玲,等. 携带Tum5基因的慢病毒表达载体的构建及其在人脐静脉血管内皮细胞中的表达[J]. 国际眼科杂志,2008,8(1):53-55.
19.	Ferrara N, Davis-Smyth T. The biology of vascular endothelial growth factor[J]. Endocr Rev,1997,18(1):4-25.
20.	Pierce EA, Avery RL, Foley ED, et al. Vascular endothelial growth factor/vascular permeability factor expression in a mouse model of retinal neovascularization[J]. Proc Natl Acad Sci USA,1995,92(3):905-909.

1. Ye HX, Yao Y, Jiang XJ, et al. Tumstatin transfected into human glioma cell line U251 represses tumor growth by inhibiting angiogenesis[J]. Chin Med J (Engl),2013,126(9):1720-1725.
2. You Y, Xue X, Li M, et al. Inhibition effect of pcDNA-tum-5 on the growth of S180 tumor[J]. Cytotechnology,2008,56(2):97-104.
3. Thevenard J, Ramont L, Devy J, et al. The YSNSG cyclopeptide derived from tumstatin inhibits tumor angiogenesis by down-regulating endothelial cell migration[J]. Int J Cancer,2010,126(5):1055-1066.
4. 黄敏丽,罗国容,陈维平,等. Tumstatin肽对视网膜微血管内皮细胞增生和迁移的影响[J]. 眼科新进展,2008,28(9):649-652.
5. Esipov R, Beyrakhova K, Likhvantseva V, et al. Antiangiogenic and antivascular effects of a recombinant tumstatin-derived peptide in a corneal neovascularization model[J]. Biochimie,2012,94(6):1368-1375.
6. Maeshima Y, Sudhakar A, Lively JC, et al. Tumstatin, an endothelial cell-specific inhibitor of protein synthesis[J]. Science,2002,295(5552):140-143.
7. Maeshima Y, Colorado PC, Torre A,et al. Distinct antitumor properties of a type Ⅳ collagen domain derived from basement membrane[J]. J Biol Chem, 2000, 275(28):21340-21348.
8. Ricci B, Lepore D, Iossa M, et al. Effect of light on oxygen-induced retinopathy in the rat model: light and OIR in the rat[J]. Doc Ophthalmol, 1990, 74(4):287-301.
9. Smith LE, Wesolowski E, Mclellan A, et al. Oxygen-induced retinopathy in the mouse[J]. Invest Ophthalmol Vis Sci,1994,35(1):101-111.
10. Li S, Li T, Luo Y, et al. Retro-orbital injection of FITC-dextran is an effective and economical method for observing mouse retinal vessels[J]. Mol Vis, 2011,17:3566-3573.
11. Maeshima Y, Manfredi M, Reimer C, et al. Identification of the anti-angiogenic site within vascular basement membrane-derived Tumstatin[J]. J Biol Chem, 2001,276(18):15240-15248.
12. Maeshima Y, Yerramalla UL, Dhanabal M,et al.Extracellular matrix-derived peptide binds to alpha vbeta 3 integrin and inhibits angiogenesis[J].J Biol Chem,2001,276(34):31959-31968.
13. Wang L, Shi P, Xu Z, et al. Up-regulation of VEGF by retinoic acid during hyperoxia prevents retinal neovascularization and retinopathy[J]. Invest Ophthalmol Vis Sci,2014,55(7):4276-4287.
14. Ameri H, Liu H, Liu R, et al. TWEAK/Fn14 pathway is a novel mediator of retinal neovascularization[J]. Invest Ophthalmol Vis Sci,2014,55(2):801-813.
15. Li Z, He T, Du K, et al. Inhibition of oxygen-induced ischemic retinal neovascularization with adenoviral 15-lipoxygenase-1 gene transfer via up-regulation of PPAR-γ and Down-regulation of VEGFR-2 expression[J/OL]. PLoS ONE, 2014,9(1):85824[2014-01-21]. http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0085824.
16. Liu G, Wu J, Tsai H, et al. Proopiomelanocortin gene delivery induces apoptosis in melanoma through NADPH oxidase 4-mediated ROS generation[J].Free Radic Biol Med, 2014,70:14-22.
17. Maclaren RE, Groppe M, Barnard AR, et al. Retinal gene therapy in patients with choroideremia: initial findings from a phase 1/2 clinical trial[J]. Lancet, 2014,383(9923):1129-1137.
18. 王爱媛,高殿文,王桂玲,等. 携带Tum5基因的慢病毒表达载体的构建及其在人脐静脉血管内皮细胞中的表达[J]. 国际眼科杂志,2008,8(1):53-55.
19. Ferrara N, Davis-Smyth T. The biology of vascular endothelial growth factor[J]. Endocr Rev,1997,18(1):4-25.
20. Pierce EA, Avery RL, Foley ED, et al. Vascular endothelial growth factor/vascular permeability factor expression in a mouse model of retinal neovascularization[J]. Proc Natl Acad Sci USA,1995,92(3):905-909.

Chinese Journal of Ocular Fundus Diseases

Adenovirus-mediated intravitreal delivery of Tum5 inhibits retinal neovascularization in a mouse model of oxygen-induced retinopathy

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