Differential Expression of Vascular Endothelial Growth Factor-a Isoforms in Neovascular Age-Related Macular Degeneration

Grisanti, Swaantje; Zhu, Qinqing; Tatar, Olcay; Lueke, Julia; Lueke, Matthias; Tura, Aysegül; Grisanti, Salvatore

doi:10.1097/iae.0000000000000385

Cited by 19 publications

(18 citation statements)

References 30 publications

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“…Alternatively spliced transcript variants give rise to at least seven homodimeric isoforms, of which VEGF 206 , VEGF 189 , VEGF 165 , and VEGF 121 represent the major isoforms and are differentially expressed in ocular neovascular conditions. 21 Exons 1 through 5 and 8 are conserved in all isoforms except VEGF 148 , and encode the receptor-binding domain. 22 To enable genomic disruption of all VEGF isoforms, we designed CRISPR gRNAs to target locations in exon 1 of the human VEGF-A gene ( Supplemental Figs.…”

Section: Resultsmentioning

confidence: 99%

Genomic Disruption of VEGF-A Expression in Human Retinal Pigment Epithelial Cells Using CRISPR-Cas9 Endonuclease

Yiu

Tieu

Nguyen

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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PurposeTo employ type II clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 endonuclease to suppress ocular angiogenesis by genomic disruption of VEGF-A in human RPE cells.MethodsCRISPR sequences targeting exon 1 of human VEGF-A were computationally identified based on predicted Cas9 on- and off-target probabilities. Single guide RNA (gRNA) cassettes with these target sequences were cloned into lentiviral vectors encoding the Streptococcus pyogenes Cas9 endonuclease (SpCas9) gene. The lentiviral vectors were used to infect ARPE-19 cells, a human RPE cell line. Frequency of insertion or deletion (indel) mutations was assessed by T7 endonuclease 1 mismatch detection assay; mRNA levels were assessed with quantitative real-time PCR; and VEGF-A protein levels were determined by ELISA. In vitro angiogenesis was measured using an endothelial cell tube formation assay.ResultsFive gRNAs targeting VEGF-A were selected based on the highest predicted on-target probabilities, lowest off-target probabilities, or combined average of both scores. Lentiviral delivery of the top-scoring gRNAs with SpCas9 resulted in indel formation in the VEGF-A gene at frequencies up to 37.0% ± 4.0% with corresponding decreases in secreted VEGF-A protein up to 41.2% ± 7.4% (P < 0.001), and reduction of endothelial tube formation up to 39.4% ± 9.8% (P = 0.02). No significant indel formation in the top three putative off-target sites tested was detected.ConclusionsThe CRISPR-Cas9 endonuclease system may reduce VEGF-A secretion from human RPE cells and suppress angiogenesis, supporting the possibility of employing gene editing for antiangiogenesis therapy in ocular diseases.

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Section: Resultsmentioning

confidence: 99%

Genomic Disruption of VEGF-A Expression in Human Retinal Pigment Epithelial Cells Using CRISPR-Cas9 Endonuclease

Yiu

Tieu

Nguyen

et al. 2016

Invest. Ophthalmol. Vis. Sci.

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“… 13 Receptor VEGFR-2, plays important role in angiogenesis of endothelial cells amongst other receptors. 14 , 15 …”

Section: Introductionmentioning

confidence: 99%

“…VEGF-A165 in AMD cases. 19 The expression of VEGF-A could progressively affect the function of RPE cells in the mice. Mice with increased VEGF-A were found with abnormal morphology of RPE cells, reduced levels of retinal rhodopsins and impaired transport of retinoic acid between RPE and photoreceptors.…”

Section: Introductionmentioning

confidence: 99%

Exploring the role of VEGF in Indian Age related macular degeneration

Sharma

Singh

et al. 2015

ANS

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BackgroundAge related macular degeneration (AMD) is major devastating neurodegenerative disorder characterized by progressive irreversible vision loss in the elderly persons. In spite of several genetic and environmental factors, the role of VEGF and CFH predispose the pathological phenomenon in the AMD patients.PurposeThe aim of the study was to estimate the VEGF levels in the serum of AMD patients and its correlation with co-morbidity of the participants.MethodsThe study recruited the 98 AMD patients and 59 controls with proper consent of the participants as per the exclusion-inclusion criteria. The co-morbidity and socio-economic details were obtained by introducing the standard questionnaire amongst the participants. Serum levels of vascular endothelial growth factor (VEGF) was estimated by ELISA and compared with the control population of the study. The levels of VEGF in the serum of AMD patients and controls were compared with Mann-Whitney U-test. Kruskal Wallis one-way analysis of variance (ANOVA) was employed to analyze more than two variables in the study.ResultsElevated level of VEGF was found in AMD patients as compared to controls. Surprisingly, we did not find significant changes among wet AMD subtypes i.e. minimal, predominant and classic wet AMD. However, we have demonstrated the intravitreal anti-VEGF treatment (avastin) in AMD patients could reduce the systemic VEGF levels although it was not significant. Moreover, the heart ailment in the AMD patients could also influence the VEGF levels.ConclusionOur study is consistent with previous studies describing the imperative significance of VEGF in AMD pathology. However, our study did not reveal the role of VEGF in wet AMD progression but it is well established causative agent for the same. The increased levels of VEGF in heart ailment among AMD patients are significant.

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“…VEGF, an important inducer of vascular permeability and a central regulator of new blood vessel growth, is remarkably involved in choroid neovascularization formation in wet AMD (Grisanti et al, ). In this process, it has been proposed that both oxidative and inflammatory state of RPE may be also implicated (Fang, Yang, & Yang, ).…”

Section: Resultsmentioning

confidence: 99%

“…VEGF, an important inducer of vascular permeability and a central regulator of new blood vessel growth, is remarkably involved in choroid neovascularization formation in wet AMD (Grisanti et al, 2015). In this process, it has been proposed that both oxidative and F I G U R E 3 Gene expression of IL-1β (a), IL-6 (b), TNF-α (c), and IFNγ (d) relative to untreated cells (control) after the treatment of ARPE-19 cells with GPETAFLR at 50 and 100 μg/ml for 24 hr.…”

Section: Re Sults and Discussionmentioning

confidence: 99%

GPETAFLR, a biopeptide from Lupinus angustifolius L., protects against oxidative and inflammatory damage in retinal pigment epithelium cells

et al. 2019

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GPETAFLR, an octapeptide released from the enzymatic hydrolysis of lupine (Lupinus angustifolius L.) protein, has demonstrated anti‐inflammatory effect in myeloid lineage. This work aims to evaluate in retinal pigment epithelium (RPE) cells the protective role of GPETAFLR on both oxidative and inflammatory markers known to be involved in age‐related macular degeneration (AMD). In comparison with stimulated control cells, GPETAFLR increased glutathione production and diminished the secretion and gene expression of VEFG, IL‐1β, IL‐6, IFNγ, and TNF‐α, as well as reactive oxygen species, and nitrite output. Our findings reveal that GPETAFLR, a novel plant peptide, is able to protect against RPE oxidative stress and inflammation. Taken together, these results strongly support innovative nutritional strategies considering Lupinus angustifolius L. as source of proteins to prevent the onset and progression of AMD. Practical applications We reveal a novel nutraceutical impact of GPETAFLR peptide in human RPE cells to prevent oxidative and inflammatory mediators. Our results support that the intake of Lupine angustifolius L., proposed to be a reservoir of GPETAFLR, could lessen the functional decay of RPE cells, leading therefore to a slowdown of the progress of AMD during age. Not only this work, but also future simple clinical studies should raise new nutritional strategies focused on understanding the etiological role of the foods, nutrition, and metabolism in the pathogenesis of ocular disorders.

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Differential Expression of Vascular Endothelial Growth Factor-a Isoforms in Neovascular Age-Related Macular Degeneration

Cited by 19 publications

References 30 publications

Genomic Disruption of VEGF-A Expression in Human Retinal Pigment Epithelial Cells Using CRISPR-Cas9 Endonuclease

Genomic Disruption of VEGF-A Expression in Human Retinal Pigment Epithelial Cells Using CRISPR-Cas9 Endonuclease

Exploring the role of VEGF in Indian Age related macular degeneration

GPETAFLR, a biopeptide from Lupinus angustifolius L., protects against oxidative and inflammatory damage in retinal pigment epithelium cells

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