AAV-mediated gene delivery of the calreticulin anti-angiogenic domain inhibits ocular neovascularization

• Published in: Angiogenesis (London) 2018-02, Vol.21 (1), p.95-109
• Main Authors: Tu, Leilei, Wang, Jiang-Hui, Barathi, Veluchamy A., Prea, Selwyn M., He, Zheng, Lee, Jia Hui, Bender, James, King, Anna E., Logan, Grant J., Alexander, Ian E., Bee, Youn-Shen, Tai, Ming-Hong, Dusting, Gregory J., Bui, Bang V., Zhong, Jingxiang, Liu, Guei-Sheung
• Format: Article
• Language: English
• Subjects: Age; Angiogenesis inhibitors; Angiogenesis Inhibitors - biosynthesis; Angiogenesis Inhibitors - genetics; Angiography; Animal models; Animals; Antiangiogenics; Biomedical and Life Sciences; Biomedicine; Calreticulin; Calreticulin - biosynthesis; Calreticulin - genetics; Cancer Research; Cardiology; Cell Biology; Dependovirus; Diabetes mellitus; Diabetic retinopathy; Diabetic Retinopathy - diagnostic imaging; Diabetic Retinopathy - genetics; Diabetic Retinopathy - metabolism; Diabetic Retinopathy - physiopathology; Electroretinography; Endothelial cells; Eye (anatomy); Female; Fluorescein; Gene transfer; Genetic Vectors; HEK293 Cells; Human Umbilical Vein Endothelial Cells - metabolism; Human Umbilical Vein Endothelial Cells - pathology; Humans; Ischemia; Luminescent Proteins - biosynthesis; Luminescent Proteins - genetics; Macular degeneration; Macular Degeneration - diagnostic imaging; Macular Degeneration - genetics; Macular Degeneration - metabolism; Macular Degeneration - physiopathology; Mice; Oncology; Ophthalmology; Optical Coherence Tomography; Original Paper; Rats; Rats, Sprague-Dawley; Red Fluorescent Protein; Reduction; Retina; Retinal Neovascularization - diagnostic imaging; Retinal Neovascularization - genetics; Retinal Neovascularization - metabolism; Retinal Neovascularization - physiopathology; Retinopathy; Structure-function relationships; Tomography, Optical Coherence; Transduction, Genetic; Umbilical vein; Vascularization; Viruses
• ISSN: 0969-6970; 1573-7209
• DOI: 10.1007/s10456-017-9591-4

Ocular neovascularization is a common pathological feature in diabetic retinopathy and neovascular age-related macular degeneration that can lead to severe vision loss. We evaluated the therapeutic efficacy of a novel endogenous inhibitor of angiogenesis, the calreticulin anti-angiogenic domain (CAD180), and its functional 112-residue fragment, CAD-like peptide 112 (CAD112), delivered using a self-complementary adeno-associated virus serotype 2 (scAAV2) in rodent models of oxygen-induced retinopathy and laser-induced choroidal neovascularization. The expression of CAD180 and CAD112 was elevated in human umbilical vein endothelial cells transduced with scAAV2-CAD180 or scAAV2-CAD112, respectively, and both inhibited angiogenic activity in vitro. Intravitreal gene delivery of scAAV2-CAD180 or scAAV2-CAD112 significantly inhibited ischemia-induced retinal neovascularization in rat eyes (CAD180: 52.7% reduction; CAD112: 49.2% reduction) compared to scAAV2-mCherry, as measured in retinal flatmounts stained with isolectin B4. Moreover, the retinal structure and function were unaffected by scAAV2-CAD180 or scAAV2-CAD112, as measured by optical coherence tomography and electroretinography. Moreover, subretinal delivery of scAAV2-CAD180 or scAAV2-CAD112 significantly attenuated laser-induced choroidal neovascularization in mouse eyes compared to scAAV2-mCherry, as measured by fundus fluorescein angiography (CAD180: 62.4% reduction; CAD112: 57.5% reduction) and choroidal flatmounts (CAD180: 40.21% reduction; CAD112: 43.03% reduction). Gene delivery using scAAV2-CAD180 or scAAV2-CAD112 has significant potential as a therapeutic option for the management of ocular neovascularization.