Hypoxia-Inducible Factor-1α Is Associated With Sprouting Angiogenesis in the Murine Laser-Induced Choroidal Neovascularization Model

André, Helder; Tunik, Selcuk; Aronsson, Monica; Kvanta, Anders

doi:10.1167/iovs.15-16476

Cited by 47 publications

(57 citation statements)

References 45 publications

(72 reference statements)

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“…Markers for vascular pericytes and endothelial cells revealed that the newly formed vessels were NG2–negative while PECAM-1– and isolectin–positive. Animal models of induced sprouting angiogenesis in adult have determined pericyte recruitment as early as 7 days after induction of the model [20,21]. The finding of NG2–negative vascular sprouts in the presented iris model could be in agreement with ocular vascular development at P27.5, where new blood vessels, possibly anastomosis between arteries and veins, are still being formed.…”

Section: Discussionsupporting

confidence: 56%

A novel in vivo model of puncture-induced iris neovascularization

et al. 2017

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PurposeTo assess iris neovascularization by uveal puncture of the mouse eye and determine the role of angiogenic factors during iris neovascularization.MethodsUveal punctures were performed on BalbC mouse eyes to induce iris angiogenesis. VEGF-blockage was used as an anti-angiogenic treatment, while normoxia- and hypoxia-conditioned media from retinal pigment epithelium (RPE) cells was used as an angiogenic-inducer in this model. Iris vasculature was determined in vivo by noninvasive methods. Iris blood vessels were stained for platelet endothelial cell adhesion molecule-1 and vascular sprouts were counted as markers of angiogenesis. Expression of angiogenic and inflammatory factors in the puncture-induced model were determined by qPCR and western blot.ResultsPunctures led to increased neovascularization and sprouting of the iris. qPCR and protein analysis showed an increase of angiogenic factors, particularly in the plasminogen-activating receptor and inflammatory systems. VEGF-blockage partly reduced iris neovascularization, and treatment with hypoxia-conditioned RPE medium led to a statistically significant increase in iris neovascularization.ConclusionsThis study presents the first evidence of a puncture-induced iris angiogenesis model in the mouse. In a broader context, this novel in vivo model of neovascularization has the potential for noninvasive evaluation of angiogenesis modulating substances.

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Section: Discussionsupporting

confidence: 56%

A novel in vivo model of puncture-induced iris neovascularization

et al. 2017

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“…To show that CjCas9 can be expressed via AAV in other tissues such as retina in mice and to investigate the therapeutic potential of CjCas9-mediated gene surgery for the treatment of age-related macular degeneration (AMD), a leading cause of blindness in adults, we prepared an AAV9 vector encoding CjCas9 under the control of the elongation factor-1 short (EFS) promoter, enhanced green fluorescent protein (eGFP) linked to the C terminus of CjCas9 with the self-cleaving T2A peptide, and a U6 promoter-driven sgRNA specific to the Vegfa or Hif1a gene, whose expression in the retina is associated with choroidal neovascularization (CNV)2627 (Fig. 5a,b).…”

Section: Resultsmentioning

confidence: 99%

In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni

et al. 2017

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Several CRISPR-Cas9 orthologues have been used for genome editing. Here, we present the smallest Cas9 orthologue characterized to date, derived from Campylobacter jejuni (CjCas9), for efficient genome editing in vivo. After determining protospacer-adjacent motif (PAM) sequences and optimizing single-guide RNA (sgRNA) length, we package the CjCas9 gene, its sgRNA sequence, and a marker gene in an all-in-one adeno-associated virus (AAV) vector and produce the resulting virus at a high titer. CjCas9 is highly specific, cleaving only a limited number of sites in the human or mouse genome. CjCas9, delivered via AAV, induces targeted mutations at high frequencies in mouse muscle cells or retinal pigment epithelium (RPE) cells. Furthermore, CjCas9 targeted to the Vegfa or Hif1a gene in RPE cells reduces the size of laser-induced choroidal neovascularization, suggesting that in vivo genome editing with CjCas9 is a new option for the treatment of age-related macular degeneration.

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“…Hypoxia is a well‐characterized angiogenesis inducer in the eye (Ten & Pinsky ; André et al. ; Campochiaro ). To determine differential response to hypoxia, we exposed CEC or REC to 6, 12 and 24 hr of hypoxic conditions and analysed HIF‐1α and HIF‐2α protein expression.…”

Section: Resultsmentioning

confidence: 99%

“…; Ten & Pinsky ; André et al. ; Campochiaro ). It has been shown that regulation of factors as a response to hypoxia is mainly driven by hypoxia‐inducible factor (HIF) molecules (Zimna & Kurpisz ).…”

Section: Introductionmentioning

confidence: 99%