Abstract:
Purpose:Vascular endothelial growth factor receptor 2 (VEGFR2) plays a key role in VEGF-induced angiogenesis. The goal of this project was to test the hypothesis that editing genomic VEGFR2 loci using the technology of clustered regularly interspaced palindromic repeats (CRISPR)-associated DNA endonuclease (Cas)9 in Streptococcus pyogenes (SpCas9) was able to block VEGF-induced activation of Akt and tube formation. Methods:Four 20 nucleotides for synthesizing single-guide RNAs based on human genomic VEGFR2 exon 3 loci were selected and cloned into a lentiCRISPR v2 vector, respectively. The DNA fragments from the genomic VEGFR2 exon 3 of transduced primary human retinal microvascular endothelial cells (HRECs) were analyzed by Sanger DNA sequencing, surveyor nuclease assay, and next-generation sequencing (NGS). In the transduced cells, expression of VEGFR2 and VEGF-stimulated signaling events (e.g., Akt phosphorylation) were determined by Western blot analyses; VEGF-induced cellular responses (proliferation, migration, and tube formation) were examined. Results:In the VEGFR2-sgRNA/SpCas9-transduced HRECs, Sanger DNA sequencing indicated that there were mutations, and NGS demonstrated that there were 83.57% insertion and deletions in the genomic VEGFR2 locus; expression of VEGFR2 was depleted in the VEGFR2-sgRNA/SpCas9-transduced HRECs. In addition, there were lower levels of Akt phosphorylation in HRECs with VEGFR2-sgRNA/SpCas9 than those with LacZ-sgRNA/SpCas9, and there was less VEGF-stimulated Akt activation, proliferation, migration, or tube formation in the VEGFR2-depleted HRECs than those treated with aflibercept or ranibizumab. Conclusions:The CRISPR-SpCas9 technology is a potential novel approach to prevention of pathologic angiogenesis.
journal_name
Invest Ophthalmol Vis Scijournal_title
Investigative ophthalmology & visual scienceauthors
Huang X,Zhou G,Wu W,Ma G,D'Amore PA,Mukai S,Lei Hdoi
10.1167/iovs.16-20537subject
Has Abstractpub_date
2017-02-01 00:00:00pages
1228-1236issue
2eissn
0146-0404issn
1552-5783pii
2606594journal_volume
58pub_type
杂志文章abstract:PURPOSE:To determine protective properties of recombinant TCR ligands (RTLs) as a new treatment for experimental autoimmune anterior uveitis (AU). RTLs comprise the rat RT1.B beta1alpha1 domains, linked either to the guinea pig MBP69-89 peptide (RTL201), to the corresponding rat MBP69-89 peptide (RTL200), or to the car...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-1242
更新日期:2006-06-01 00:00:00
abstract:PURPOSE:To evaluate the rate and location of visual field (VF) progression before and after detection of disc hemorrhage (DH). METHODS:Disc photographs of consecutive patients with glaucoma with >or=5 SITA-Standard 24-2 VF in either eye were evaluated for the presence and location of DH. Exclusion criteria included di...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.09-3446
更新日期:2009-10-01 00:00:00
abstract:Purpose:Although normal function of the lacrimal gland is essential for vision (and thus for human well-being), the lacrimal gland remains rather poorly understood at a molecular level. The purpose of this study was to identify new genes and signaling cascades involved in lacrimal gland development. Methods:To identif...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.16-20315
更新日期:2017-02-01 00:00:00
abstract:Purpose:The purpose of this study was to evaluate macular preganglionic function and to verify its relationship with retinal and choroidal morphology in patients with intermediate age-related macular degeneration (iAMD) patients. Methods:All included patients performed multifocal electroretinogram (mfERG) for investig...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.61.5.11
更新日期:2020-05-11 00:00:00
abstract:PURPOSE:To compare the diagnostic value of Bruch's membrane opening minimum rim width (BMO-MRW) and retinal nerve fiber layer thickness (RNFLT) in patients with ocular hypertension, preperimetric, and perimetric glaucoma. METHODS:One hundred eighty-one eyes consisting of 40 healthy controls, 41 ocular hypertensive sub...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章,随机对照试验
doi:10.1167/iovs.15-18906
更新日期:2016-07-01 00:00:00
abstract:PURPOSE:To determine whether polyamines are present in corneal cells, whether corneal cell polyamines can be depleted by blocking the first rate-limiting enzyme in the polyamine synthesis pathway, ornithine decarboxylase (ODC), and whether polyamines are required for proliferation in all three corneal cell types. METH...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.02-0889
更新日期:2003-06-01 00:00:00
abstract:PURPOSE:Bevacizumab eyedrops inhibit corneal neovascularization. The purpose of this study was to analyze the safety profile of VEGF-A neutralization at the ocular surface. METHODS:Bevacizumab eyedrops (5 mg/mL) and an antimurine VEGF-A antibody (250 microg/mL) were applied to normal murine corneas five times a day fo...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.07-1129
更新日期:2009-05-01 00:00:00
abstract:PURPOSE:The aim of this study was to use Raman spectra of tears to differentiate between infectious and noninfectious ulcerative keratitis. METHODS:Raman microspectroscopy was applied using the drop-coating deposition method on Ti/Au-coated glass slides to obtain sample spectra from different human tear groups, includ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.11-7923
更新日期:2012-03-15 00:00:00
abstract:PURPOSE:The S1 RNA binding domain 1 (SRBD1) and elongation of long-chain fatty acids family member 5 (ELOVL5) have been reported to be susceptibility genes for early-onset normal-tension glaucoma (NTG). The present study we conducted to assess whether these genes were associated with primary open-angle glaucoma (POAG),...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章,多中心研究
doi:10.1167/iovs.11-7382
更新日期:2011-06-28 00:00:00
abstract:PURPOSE:To investigate the choroidal thickness (CT) in children with amblyopia through spectral-domain optical coherence tomography (SD-OCT). METHODS:Thirty-seven children with unilateral amblyopia and 22 children with normal vision participated in the study. Cross-sectional images of the choroid of evaluated eyes wer...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.14-14439
更新日期:2014-10-14 00:00:00
abstract:PURPOSE:Vasoactive intestinal peptide (VIP) is expressed by corneal endothelial (CE) cells and is present in the aqueous humor, which bathes CE cells in vivo. This study demonstrated the role of CE cell VIP in maintaining the expression level of a CE differentiation marker, N-cadherin, and the hexagonal cell shape. ME...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.07-1543
更新日期:2008-08-01 00:00:00
abstract:Purpose:To employ quantitative fundus autofluorescence (qAF) imaging in rhesus macaques to noninvasively assess retinal pigment epithelial (RPE) lipofuscin in nonhuman primates (NHPs) as a model of aging and age-related macular degeneration (AMD). Methods:The qAF imaging was performed on eyes of 26 rhesus macaques (me...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.61.8.16
更新日期:2020-07-01 00:00:00
abstract:PURPOSE:Recent studies on ocular shape have raised increased interest in the peripheral characteristics of the eye, as it potentially triggers changes in the central vision. Current techniques are, however, not capable of accurately measuring the three-dimensional shape of the retina. We describe a new magnetic resonan...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.14-15161
更新日期:2015-01-15 00:00:00
abstract:PURPOSE:To identify the localization of indoleamine 2,3-dioxygenase (IDO) in human corneal cells and to evaluate its functional ability as a local immunosuppressive factor. METHODS:The expression profile of IDO was identified in primary cultures of human corneal cells (fibroblasts, epithelial cells, endothelial cells)...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-1336
更新日期:2007-09-01 00:00:00
abstract:PURPOSE:Severe ocular allergic diseases are characterized by pronounced conjunctival inflammation triggered by T helper 2 (Th2) cells and corneal epithelial damage induced by eosinophils. To examine the role of alarmins released by damaged corneal epithelial cells in tissue eosinophilia, we investigated the effects of ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.13-11969
更新日期:2013-08-27 00:00:00
abstract::The definition and diagnostic criteria of dry eye disease (DED) proposed by the Japan Dry Eye Society and other countries are reviewed. The first definition and criteria of DED in Japan were proposed in 1995. In that report, DED was considered a disease of tears, which subsequently damaged ocular epithelia. The presen...
journal_title:Investigative ophthalmology & visual science
pub_type: 历史文章,杂志文章,评审
doi:10.1167/iovs.17-23475
更新日期:2018-11-01 00:00:00
abstract:PURPOSE:People with limited literacy are at increased risks of chronic systemic conditions. The authors therefore investigated the independent contribution of limited literacy on visual impairment and visual function in a large eye survey in Singapore. METHODS:The authors undertook a population-based, cross-sectional ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.11-7725
更新日期:2011-09-29 00:00:00
abstract:PURPOSE:The RAS association domain family 1 (RASSF1) gene is a tumor-suppressor gene located on chromosome 3p21.3. The alternative transcript (RASSF1a) has been shown to be inactivated by hypermethylation in several human malignancies, including breast, prostate, and lung cancer, and in cutaneous melanoma. The purpose ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.06-0781
更新日期:2007-02-01 00:00:00
abstract:PURPOSE:There is uncertainty in the published literature as to whether physical activity should be advocated for age-related macular degeneration (AMD) prevention. We aimed to assess prospectively the association between physical activity and the 15-year incidence of AMD in older adults. METHODS:We assessed AMD from r...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.14-15575
更新日期:2014-11-11 00:00:00
abstract::The rate of aqueous humor flow of 19 normal subjects was measured by a fluorescein clearance technique during the day, during sleep (at night) and during sleep deprivation (at night). Subjects engaged in routine activities during the daytime measurements and slept in comfortable quarters during the nighttime measureme...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1984-06-01 00:00:00
abstract:PURPOSE:Angiotensin-converting-enzyme (ACE) catalyses the formation of angiotensin II (ANGII), which presumably acts as a central neurotransmitter/modulator. ANGII-related effects have also been observed in the retina. Present in vivo experiments were designed to investigate further ANGII-related effects on retinal neu...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1994-03-01 00:00:00
abstract:PURPOSE:Visual sensations in patients with blindness and retinal degenerations may be restored by electrical stimulation of retinal neurons with implantable microelectrode arrays. A prospective trial was initiated to evaluate the safety and efficacy of a wireless intraocular retinal implant (EPIRET3) in six volunteers ...
journal_title:Investigative ophthalmology & visual science
pub_type: 临床试验,杂志文章
doi:10.1167/iovs.08-2752
更新日期:2009-06-01 00:00:00
abstract:PURPOSE:To investigate a depolarizing layer that is visible in polarization-sensitive optical coherence tomography (PS-OCT) images of the retina. To identify this layer and characterize its depolarizing effect quantitatively. METHODS:Ten healthy human subjects (mean age, 31 +/- 8 years) and two patients with RPE disea...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-1589
更新日期:2006-12-01 00:00:00
abstract:PURPOSE:To compare the clinical outcomes of aberration-free ablation profiles based on the normal corneal vertex (CV) and the pupil center (PC) in relation to laser in situ keratomileusis. Aberration-free aspheric ablation treatments were performed in all cases. METHODS:Two myopic astigmatism groups (CV centered using...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.08-2176
更新日期:2008-12-01 00:00:00
abstract:PURPOSE:Retinal pigment epithelial (RPE) cells play important roles in ophthalmologic diseases such as proliferative vitreoretinopathy, AMD, and diabetic retinopathy. MicroRNA-34a (miR-34a) has been reported to be important in the regulation of cell proliferation, migration, differentiation, and apoptosis. In this stud...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.13-11873
更新日期:2013-10-03 00:00:00
abstract:PURPOSE:This study was performed to investigate motion perception in patients with glaucoma. METHODS:A random dot motion test was used to measure three aspects of central motion perception: minimum displacement threshold (Dmin), maximum displacement threshold (Dmax), and coherence threshold (signal to noise). Motion p...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1993-12-01 00:00:00
abstract:PURPOSE:In a group of humans with strabismic amblyopia, the relationship was examined between the structure and function of different brain regions. Three question were addressed: (1) Is the lateral geniculate nucleus (LGN) in humans with amblyopia structurally as well as functionally abnormal? (2) Do structural anomal...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.09-3931
更新日期:2010-03-01 00:00:00
abstract:PURPOSE:Recent studies suggest that photoreceptor cells regulate local inflammation in the retina in diabetes. The purpose of this study was to determine if photoreceptor cells themselves produce inflammatory proteins in diabetes and if soluble factors released by photoreceptors in elevated glucose induce inflammatory ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.16-19859
更新日期:2016-08-01 00:00:00
abstract:PURPOSE:To investigate the effect of amino acids on ornithine cytotoxicity in ornithine-delta-aminotransferase (OAT)-deficient human retinal pigment epithelial (RPE) cells as an in vitro model of gyrate atrophy (GA) of the choroid and retina. METHODS:RPE cells were treated with 0.5 mM 5-fluoromethylornithine (5-FMOrn)...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.03-0403
更新日期:2003-11-01 00:00:00
abstract:PURPOSE:To identify if whole-body hydration plays an important role in dry eye (DE). We hypothesized that individuals classified as DE have higher plasma osmolality (Posm), indicating suboptimal hydration, compared with those classified as non-DE. METHODS:Using a hospital-based observational cross-sectional design, as...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.12-10175
更新日期:2012-09-25 00:00:00