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Tetrahedral framework nucleic acids-based delivery of microRNA-155 inhibits Choroidal neovascularization by regulating the polarization of macrophages

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Bioactive Materials 2022年第8期7卷 134-144页

作者： Xin Qin Lirong Xiao Ni Li Chen Hou Wenman Li Jiajie Li Naihong Yan Yunfeng Lin State Key Laboratory of Oral Diseases National Clinical Research Center for Oral DiseasesWest China Hospital of StomatologySichuan UniversityChengdu610041China Research Laboratory of Ophthalmology West China HospitalSichuan UniversityChengdu610041China College of Biomedical Engineering Sichuan UniversityChengdu610041China

Choroidal neovascularization(CNV)is a common pathological feature of various eye diseases and an important cause of visual impairment in middle-aged and elderly patients.In previous studies,tetrahedral framework nucleic acids(tFNAs)showed good carrier performance.In this experiment,we developed microRNA-155-equipped tFNAs(T-155)and explored its biological effects on CNV.Based on the results of in-vitro experiments,T-155 could regulate macrophages into the antiangiogenic M1 type.Then,we injected T-155 into the vitreous of laser-induced CNV model mice and found that T-155 significantly reduced the size and area of CNV,inhibited blood vessel leakage.In summary,we prove that T-155 could regulate the inflammatory process of CNV by polarizing macrophages,thereby improving the symptoms of CNV.Thus,T-155 might become a new DNA-based drug with great potential for treating CNV.

关键词： Choroidal neovascularization Tetrahedral framework nucleic acid Macrophages MicroRNA-155 Immunity therapy

Effect of integrin α5β1 inhibition on SDF-l/CXCR4-mediated Choroidal neovascularization

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International Journal of Ophthalmology(English edition) 2018年第5期11卷 726-735页

作者： Yang Lyu Wen-Qin Xu Li-Juan Sun Xiao-Yan Pan Jian Zhang Yu-Sheng Wang Department of Ophthalmology Eye Institute of China PLAXijing Hospitalthe Fourth Military Medical University Department of Ophthalmology General Hospital of Lanzhou Military Command Department of Biochemistry and Molecular Biology the Fourth Military Medical University

AIM：To investigate the roles of integrins in Choroidal neovascularization（CNV） and their associations with the stromal cell-derived factor-1（SDF-1）/CXCR4 axis.METHODS：CNV lesions were induced in mice using laser photocoagulation.After CNV induction,all animals were randomly assigned to：control,SDF-1,SDF-1＋age-related macular degeneration（AMD） 3100（CXCR4 inhibitor）,and SDF-1＋ATN161（integrin α5β1 inhibitor） groups;their effects on CNV progression were observed using hematoxylin eosin（HE） staining,fundus fluorescein angiography（FFA） grading and optical coherence tomography（OCT）,and their effects on CXCR4/integrin α5 expression were evaluated using Western blot and double immunofluorescence staining.Hypoxia-exposed endothelial cells（ECs） were used to simulate CNV in vitro,they were treated with SDF-1,combined with CXCR4 siRNA/AMD3100 or ATN161,and expression of integrin α5,cell migration and tube formation were analyzed.RESULTS：Integrin subunit α5 increased at 3^ rd and 7^ th day and decreased at 14 ^th day in CNV mice,with no significant change of β1-integrin.CXCR4 expression in CNV mice had persistent increase within 14 d after induction.SDF-1 treatment significantly promoted the CNV progression during 3-14 d.The mean CNV length in AMD3100 andATN161 group at day 7 was 270.13 and 264.23 μm in HE images,significantly lower than the mean length in SDF-1（345.70 μm） group.AMD3100 and ATN161 also significantly reduced thickness and leakage of CNV induced by SDF-1.Mean integrin α5 positive area in SDF-1 group reached 2.31×104 μm^2,significantly higher than control（1.25×104 μm^2）,which decreased to 1.78×104 μm^2 after AMD3100 treatment.About 61.36% of ECs in CNV lesions expressed α5 in SDF-1 group,which significantly decreased to 43.12% after AMD3100 treatment.In vitro,integrin α5 peaked by 6 folds after 6 h of hypoxia exposure and CXCR4 gradually increased by up to 2.3 folds after 24 h of hypoxia.Approximately 25.12% of ECs expressed integrin α5 after SDF-1 stimulation,which decreased to 7.2%-9.5% after si-CXCR4 or AMD310

关键词： Choroidal neovascularization endothelial cells stromal cell-derived factor-1 CXCR4 integrin α5β1 hypoxia

Correlation of In Vivo and In Vitro Methods in Measuring choroidal Vascularization Volumes Using a Subretinal Injection Induced Choroidal neovascularization Model

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Chinese Medical Journal 2015年第11期128卷 1516-1522页

作者： Chuang Nie Mao-Nian Zhang Hong-Wei Zhao Thomas D Olsen Kyle Jackman Lian-Na Hu Wen-Ping Ma Xiao-Fei Chen Juan Wang Ying Zhang Tie-Shah Gao Hiro Uehara Balamurali K Ambat Ling Luo Department of Ophthalmology Chinese PLA General Hospital Beijing 100853 China Department of Ophthalmology The 306th Hospital of PLA Beijing 100101 China Moran Eye Center University of Utah Salt Lake City Utah 84132 USA Department of Ophthalmology The 26th Hospital of PLA Beijing 100094 China

Background： In vivo quantification of Choroidal neovascularization （CNV） based on noninvasive optical coherence tomography （OCT） examination and in vitro choroidal flatmount immunohistochemistry stained of CNV currently were used to evaluate the process and severity of age-related macular degeneration （AMD） both in human and animal studies. This study aimed to investigate the correlation between these two methods in murine CNV models induced by subretinal injection. Methods： CNV was developed in 20 C57BL6/j mice by subretinal injection of adeno-associated viral delivery of a short hairpin RNA targeting sFLT-1 （AAV.shRNA.sFLT- 1）, as reported previously. After 4 weeks, CNV was imaged by OCT and fluorescence angiography. The scaling factors for each dimension, x, y, and z （ktm/pixel） were recorded, and the corneal curvature standard was adjusted from human （7.7） to mice （1 .4）. The volume of each OCT image stack was calculated and then normalized by multiplying the number of voxels by the scaling factors for each dimension in Seg3D software （University of Utah Scientific Computing and Imaging Institute, available at http：//www.***.utah.edu/cibc-software/seg3d.html）. Eighteen mice were prepared for choroidal flatmounts and stained by CD31. The CNV volumes were calculated using scanning laser confocal microscopy after immunohistochemistry staining. Two mice were stained by Hematoxylin and Eosin for observing the CNV morphology. Results： The CNV volume calculated using OCT was, on average, 2.6 times larger than the volume calculated using the laser confocal microscopy. The correlation statistical analysis showed OCT measuring of CNV correlated significantly with the in vitro method （R： = 0.448, P = 0.001, n = 18）. The correlation coefficient for CNV quantification using OCT and confocal microscopy was 0.693 （n = 18, P = 0.001 ）. Conclusions： There is a fair linear correlation on CNV volumes between in vivo and in vitro methods in CNV models induced by subretinal injection. The result might provide a useful evaluation

关键词： choroidal Flatmount Choroidal neovascularization Correlation Spectral-domain Optical Coherence Tomography

Effects of endogenous dopamine induced by low concentration atropine eye drops on Choroidal neovascularization in high myopia mice

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International Journal of Ophthalmology(English edition) 2023年第7期16卷 1034-1040页

作者： Yan-Yan Ji Shi-Xi Zhang Ye Kang Song Chen Clinical College of Ophthalmology Tianjin Medical UniversityTianjin 300020China Tianjin Eye Hospital Tianjin 300020China Liaocheng Aier Eye Hospital Liaocheng 252100Shandong ProvinceChina Dezhou Aier Eye Hospital Dezhou 253000Shandong ProvinceChina YanTai Aier Eye Hospital Yantai 264000Shandong ProvinceChina

AIM:To evaluate effects of endogenous dopamine induced by low concentration atropine eye drops on Choroidal neovascularization(CNV)in high myopia mice.METHODS:The C57BL/6J mice were deprived of the right eye for 4wk,and the high myopia was diagnosed by optometry,the diopter was less than-6.00 D,and CNV was induced by 532 nm laser.The changes of dopamine D1 receptor(DRD1),dopamine D2 receptor(DRD2),and vascular endothelial growth factor A(VEGFA)were detected by Western blot technology at 0.5,1,2h,and 7d after 0.01%,0.05%,and 0.1%atropine eye drops,respectively,the area of CNV was measured.RESULTS:Significant increases were observed on the expression of DRD2 in mouse high myopia model at 0.5,1,2h,7d with 0.05%and 0.1%atropine eye drops(P<0.05).Significant decreases were observed on the expression of DRD1 and VEGFA in mouse high myopia model at 0.5,1,2h,7d with 0.05%and 0.1%atropine eye drops(P<0.05).The area of CNV induced by laser in the drug-treated group was significantly smaller than that in the control group,and the higher the concentration,the more significant the inhibitory effect(P<0.05).CONCLUSION:The 0.01%,0.05%,0.1%atropine eye drops can decrease the level of VEGFA and inhibit high myopia CNV indirectly by up-regulating the level of DRD2 and down-regulating the level of DRD1,and the effect of 0.05%and 0.1%atropine eye drops is more significant.

关键词： high myopia Choroidal neovascularization low concentration atropine eye drops dopamine D1 receptor dopamine D2 receptor

Effect of cytokeratin 17 on retinal pigment epithelium degeneration and Choroidal neovascularization

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International Journal of Ophthalmology(English edition) 2016年第3期9卷 363-368页

作者： Yi Shen Pei Zhuang Tao Xiao George CY Chiou Institute of Ocular Pharmacology College of Medicine Texas A&M Health Science Center College Station TX 77843USA

AIM： To study the effects of cytokeratin 17 （CK17） on sodium iodate （NalOs） induced rat retinal pigment epithelium （RPE） degeneration, laser induced rat Choroidal neovascularization （CNV）, and oxidative stress of human retinal pigment epithelium cells （ARPE-19） and human umbilical vein endothelial cell （HUVEC）. METHODS： Thirty 8-week-old male Brown Norway rats were randomly divided into 3 groups, 10 rats in control group treated with solvent alone; 10 rats in NalOs group treated with solvent and 35 mg/kg NalO3 injection through hypoglossal vein and 10 rats in CK17 ＋NaIOs group treated with 1% CK17 eye drop 3 times a day for lwk before and 4wk after NalOs injection. RPE function was measured with c-wave of electroretinogram （ERG）. Another 20 rats were randomly divided into 2 groups. Of them 10 rats in CK17 group were anesthetized to receive Nd：YAG laser and given 1% CK17 eye drop before same as above; 10 rats in control were received Nd：YAG and treated with solvent. The development of Choroidal neovascularization （CNV） was determined by fundus fluorescein angiography （FFA） performed on 4wk after laser. Methylthiazoly tetrazolium （MTT） assay was used to study effect of CK17 on various oxidants induced injury in ARPE-19 and HUVEC /n vitro RESULTS： Four weeks after NalOs injection, the c- wave amplitude of ERG was 0.393±0.02 V in the control group, 0.184±0.018 V in NalOs group and 0.3±0.01 V in CK17＋NalOs group. There was a significant reversal of the c-wave by CK17 as compared to NalOs group （P〈0.01）. Four weeks after laser, the size of the CNV lesion was 2.57±0.27 mm2 in control group and 1.64 ±0.08 mm2 in CK17 group. The lesion size significantly diminished in CK17 group （P〈0.01）. The inn vitro results showed CK17 also reversed the various oxidants induced injuries in ARPE-19 at the dose of 100 μg/mL and enhanced the injury in HUVECs at different concentrations. CONCLUSION： CK17 can significantly protect RPE from NalOs induced degeneration in vivo and /n vito and also could reverse the various oxidants induced in

关键词： cytokeratin 17 age-related maculardegeneration Choroidal neovascularization retinal pigmentepithelium human umbilical vein endothelial cells

Inhibition of Choroidal neovascularization by lentivirusmediated PEDF gene transfer in rats

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International Journal of Ophthalmology(English edition) 2016年第8期9卷 1112-1120页

作者： Ya-Jie Yu Bin Mo Lu Liu Yan-Kun Yue Chang-Li Yue Wu Liu Beijing Tongren Eye Center Beijing Tongren HospitalCapital Medical University Beijing Ophthalmology & Visual Sciences Key Laboratory Department of Ophthalmology Fuxing Hospital Capital Medical University Department of Pathology Beijing Tongren Hospital Capital Medical University

AIM: To evaluate the effects of lentivirus-mediated pigment epithelium-derived factor (PEDF) gene transfer performed in treatment of rats with established Choroidal neovascularization (CNV), and investigates the mechanism by which PEDF inhibits CNV in rats. METHODS: Brown Norway (BN) rats (n=204) were induced by exposure to a laser, and then randomly assigned to 3 groups: no treatment; treatments with intravitreal injection of lentivirus-PEDF-green fluorescent protein (GFP) or lentivirus-control GFP (free fluorescent protein). Following induction and treatment, the CNV tissue was assessed for form, size and vessel leakage by fluorescein fundus angiography (FFA), optical coherence tomography (OCT), histopathology, and examination of choroidal flat mounts. VEGF, Flk-1, and PEDF expression were evaluated by real-time polymerase chain reaction (PCR) and Western blot. RESULTS: A stable laser-induced rat model of CNV was successfully established, and used to demonstrate lentivirus-mediated REDO gene transfer by intravitreal injection. Expression of green fluorescence labelled PEDF was observed in the retina up to 28d after injection. An intravitreal injection of lentivirus-PEDF-GFP at 7d led to a significant reduction in the size, thickness and area of CNV showed by FFA, OCT and choroidal flat mounts. PEDF was up-regulated while VEGF and Flk-1 were down-regulated in the lentivirus-PEDF-GFP group. The differences in VEGF and Flk-1 expression in the control and lentivirus-PEDF groups at 7, 14, 21 and 28d after laser induction were all statistically significant. CONCLUSION: Lentivirus-mediated PEDF gene transfer is effective for use in treatment of laser-induced CNV, and PEDF exerts its therapeutic effects by inhibiting expression of VEGF and Flk-1.

关键词： pigment epithelium-derived factor Choroidal neovascularization lentivirus vascular endothelial growth factor Flk-1

Intravitreal ranibizumab for the treatment of pathological myopia associated with Choroidal neovascularization in Chinese patients Intravitreal ranibizumab for the treatment of pathological myopia associated with Choroidal neovascularization in Chinese patients

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Chinese Medical Journal 2014年第16期127卷 2906-2910页

作者： Yang Xu Dai Hong Department of Ophthalmology Beijing Hospital Beijing 100730China

Background Pathological myopia （PM） is the leading cause for Choroidal neovascularization （CNV） in people below 50 years of age,the anti-vascular endothlial growth factor （VEGF） medicine is now available to treat CNV secondary to PM.This study aimed to observe the efficacy of intravitreal ranibizumab for PM associated with subfoveal or juxtafoveal CNV in Chinese patients.Methods Fifty-four eyes of 52 consecutive patients were included,they treated with intravitreal ranibizumab 0.5 mg for PM associated with CNV.The best corrected visual acuity （BCVA） of Snellen chart,letters of ETDRS chart,retinal thickness,leakage of CNV lesion,and complications with surgery were analyzed pre-and post-treatment.Eligibility criteria included diopter ≥-8.0 D or eye axis ≥28 mm with fundus changes of PM （lacquer crack,optic disc atrophy,chorioretinal atrophy,posterior scleral staphyloma）; CNV secondary to PM; subfoveal or juxtafoveal CNV.Results For 54 affected eyes of 52 consecutive patients,the average BCVA of Snellen chart and letters of ETDRS chart were 0.29 and 30.4,respectively; fundus fluorescein angiography （FFA）/indocyanine green angiography （ICGA） showed CNV leakage,and average retinal thickness on optical coherence tomography （OCT） was 267.2 μm before treatment.Injections of ranibizumab ranged from 1 to 4 （mean 2.2）.Follow-up time varied from 12 to 36 months （mean 31.9 months）.At the last visit,the BCVA of Snellen chart was increased by three lines （mean 0.65）（P 〈0.01）; the letters of ETDRS chart were increased to 17.0 letters （mean 47.4,P 〈0.01）; the visual acuity increased more than 15 letters in 30 eyes （55.5%）,decreased in 1 eye （1.9%）; the retinal thickness on OCT images was decreased by 17.0 μm （mean 250.2 μm）（P=0.082）; no active leakage from the CNV lesion occurred in 18 eyes （33.3%）,reduced leakage in 30 eyes （55.6%）,and no change in 6 eyes （11.1%） as shown by FFA/ICGA.Increased retinoschisis was observed in one eye after the second injection.Conclusions Intravitreal ranibizumab for neovascular PM was well toler

关键词： ranibizumab pathological myopia Choroidal neovascularization

Multimodal imaging of experimental Choroidal neovascularization

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International Journal of Ophthalmology(English edition) 2022年第6期15卷 886-893页

作者： Ioanna Tsioti Xuan Liu Petra Schwarzer Martin S.Zinkernagel Despina Kokona Department of Ophthalmology InselspitalBern University Hospitaland Department of BioMedical ResearchUniversity of BernBern 3010Switzerland Department of Ophthalmology the First Affiliated Hospital of Xi’an Jiaotong UniversityXi’an 710061Shaanxi ProvinceChina

AIM:To compare Choroidal neovascularization(CNV)lesion measurements obtained by in vivo imaging modalities,with whole mount histological preparations stained with isolectin GS-IB4,using a murine laser-induced CNV model.METHODS:B6 N.Cg-Tg(Csf1 r-EGFP)1 Hume/J heterozygous adult mice were subjected to laser-induced CNV and were monitored by fluorescein angiography(FA),multicolor(MC)fundus imaging and optical coherence tomography angiography(OCTA)at day 14 after CNV induction.choroidalretinal pigment epithelium(RPE)whole mounts were prepared at the end of the experiment and were stained with isolectin GS-IB4.CNV areas were measured in all different imaging modalities at day 14 after CNV from three independent raters and were compared to choroidal-RPE whole mounts.Intraclass correlation coefficient(ICC)type 2(2-way random model)and its 95%confidence intervals(CI)were calculated to measure the correlation between different raters’measurements.Spearman’s rank correlation coefficient(Spearman’s r)was calculated for the comparison between FA,MC and OCTA data and histology data.RESULTS:FA(early and late)and MC correlates well with the CNV measurements ex vivo with FA having slightly better correlation than MC(FA early Spearman’s r=0.7642,FA late Spearman’s r=0.7097,and MC Spearman’s r=0.7418),while the interobser ver reliability was good for both techniques(FA early ICC=0.976,FA late ICC=0.964,and MC ICC=0.846).In contrast,OCTA showed a poor correlation with ex vivo measurements(Spearman’s r=0.05716)and high variability between different raters(ICC=0.603).CONCLUSION:This study suggests that FA and MC imaging could be used for the evaluation of CNV areas in vivo while caution must be taken and comparison studies should be performed when OCTA is employed as a CNV monitoring tool in small rodents.

关键词： Choroidal neovascularization in vivo imaging fluorescein angiography multicolor fundus imaging optical coherence tomography angiography

A novel mutation of CYP4V2 gene associated with Bietti crystalline dystrophy complicated by Choroidal neovascularization

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International Journal of Ophthalmology(English edition) 2022年第6期15卷 940-946页

作者： Xin-Yao Han Lin-Qi Zhang Ji-Yang Tang Lyu-Zhen Huang Ran Tang Jin-Feng Qu Department of Ophthalmology Peking University People’s HospitalEye Diseases and Optometry InstituteBeijing Key Laboratory of Diagnosis and Therapy of Retinal and Choroid DiseasesCollege of OptometryPeking University Health Science CenterBeijing 100044China

AIM: To investigate the clinical characteristics and genetic features of a Bietti crystalline dystrophy(BCD) proband in a Chinese family.METHODS: A Chinese female diagnosed with BCD complicated by bilateral Choroidal neovascularization(CNV) and her parents underwent complete ophthalmic examinations, including fundus autofluorescence(AF), fundus photography(FP), fundus fluorescein angiography(FFA), visual field testing, full-field electroretinography(ERG), optical coherence tomography(OCT) and optical coherence tomography angiography(OCTA). The sequencing of the CYP4 V2 gene was performed to the whole family.RESULTS: Bilateral tiny glittering crystal-like deposits and differing extent of atrophy of the retinal pigment epithelium(RPE) were found in the posterior pole of her fundus. The diffuse hypo-fluorescence shown on AF images and window defects shown on FFA both indicated the atrophy of the RPE and choriocapillaris. OCT showed the thinning of the RPE and choriocapillaris layer, ellipsoid zone(EZ) band defect and CNV in both eyes. OCTA images proofed bilateral type 2 CNV. The visual field test showed central and paracentral scotoma. ERG showed a slightly decreased b-wave in scotopic ERG. Gene sequencing identified three mutations of the CYP4 V2 gene, c.802_807 del, c.810 del T, and c.1388 G>A. The mutation c.1388 G>A was a novel substitution mutation.CONCLUSION: The novel mutation c.1388 G>A may be a possible cause that could induce the clinical phenotype of BCD.

关键词： Bietti crystalline dystrophy retinal degeneration CYP4V2 gene Choroidal neovascularization