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

Abstract

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.

• Authors: Leilei Tu, Jiang-Hui Wang, Veluchamy A Barathi, Selwyn M Prea, Zheng He, Jia Hui Lee, James Bender, Anna E King, Grant J Logan, Ian E Alexander, Youn-Shen Bee, Ming-Hong Tai, Gregory J Dusting, Bang V Bui, Jingxiang Zhong, Guei-Sheung Liu
• Journal: Angiogenesis (Angiogenesis) Vol. 21 Issue 1 Pg. 95-109 (02 2018) ISSN: 1573-7209 [Electronic] Germany
• PMID: 29318471 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Angiogenesis Inhibitors
• Calreticulin
• Luminescent Proteins

Topics

• Angiogenesis Inhibitors (biosynthesis, genetics)
• Angiography
• Animals
• Calreticulin (biosynthesis, genetics)
• Dependovirus
• Diabetic Retinopathy (diagnostic imaging, genetics, metabolism, physiopathology)
• Electroretinography
• Female
• Genetic Vectors
• HEK293 Cells
• Human Umbilical Vein Endothelial Cells (metabolism, pathology)
• Humans
• Luminescent Proteins (biosynthesis, genetics)
• Macular Degeneration (diagnostic imaging, genetics, metabolism, physiopathology)
• Mice
• Rats
• Rats, Sprague-Dawley
• Retinal Neovascularization (diagnostic imaging, genetics, metabolism, physiopathology)
• Tomography, Optical Coherence
• Transduction, Genetic
• Red Fluorescent Protein