RNA测序法比较POAG与非POAG供体眼球小梁网基因表达的差异  

Differential gene expression profiles of trabecular meshwork between POAG and non-POAG donated eyes by using RNA-sequencing

作　　者：刘丽芳[1] 曾锦惠 黄楚开[1] 王耿[1] 张铭志[1] Liu Lifang;Zeng Jinhui;Huang Chukai;Wang Geng;Zhang Mingzhi(Joint Shantou International Eye Center of Shantou University and the Chinese University of Hong Kong,Shantou 515041,China)

机构地区：[1]汕头大学·香港中文大学联合汕头国际眼科中心,515041

出　　处：《中华实验眼科杂志》2020年第8期646-652,共7页Chinese Journal Of Experimental Ophthalmology

基　　金：广东省自然科学基金项目(2015A030310504);汕头市医学科技人才培育及临床技术提升计划项目(汕府科[2019]106号)。

摘　　要：目的使用RNA测序技术比较原发性开角型青光眼(POAG)与正常人小梁网组织基因表达谱的差异,探讨POAG可能的基因学病因。方法实验组小梁网组织来自汕头国际眼科中心行小梁切除术的3例POAG患者,对照组小梁网组织来自汕头眼库的2例非POAG供体眼球。提取2个组小梁网组织RNA进行测序获得基因表达谱,用R软件包edgeR分析实验组与对照组基因表达的差异,用DAVID生物信息分析网站对差异基因进行基因本体(GO)及功能注释聚类分析。采用KOBAS 3.0进行PANTHER富集通路分析,揭示POAG可能的基因学病因。结果(1)RNA测序共得到28821条基因,2个组间有统计学差异的基因22条[错误发现率(FDR)<0.05],其中转录表达上升的基因1条,转录表达下降的基因21条;(2)表达差异的基因功能集中,生物学过程涉及角化、表皮发育、中间丝细胞骨架组织等,细胞成分涉及角蛋白丝、中间丝、细胞外泌体、触珠蛋白-血红蛋白复合体等,分子功能涉及结构分子活性、细胞骨架结构组成等;(3)与差异基因相关的PANTHER富集通路主要包括纤溶酶原激活级联反应、p38 MAPK、氧化应激反应及p53通路等。结论角蛋白表达异常、中间丝细胞骨架结构改变、血纤维蛋白溶酶原激活级联反应以及p38 MAPK等通路调控改变引起的小梁网及细胞外基质重塑可能是POAG发病的原因。其中,与发病机制相关的差异基因主要包括细胞骨架相关基因及细胞外基质重塑相关基因,细胞骨架及细胞外基质可作为青光眼治疗的靶组织。Objective To investigate gene basis of primary open angle glaucoma(POAG)by comparing gene expression profile of trabecular meshwork between POAG patients and normal controls by using RNA-sequencing.Methods Trabecular meshwork specimen were obtained from trabeculectomy(POAG group,n=3)or donated eyes(control group,n=2).RNA was extracted and sequenced in both groups,gene expression profiles were analyzed and compared between them,and different expression genes associated with POAG were revealed by using Database for Annotation,Visualization and Integrated Discovery(DAVID)and Protein Analysis Through Evolutionary Relationships(PANTHER)gene list analysis.Written informed consent was obtained from each patient or the family members prior to entering the study cohort.The study protocol was approved by the Ethics Committee of Joint Shantou International Eye Center of Shantou University and the Chinese University of Hong Kong[No.EC20140311(3)-P01].Results(1)Total of 28821 genes were obtained from RNA-sequencing,22 genes were statistically significant between the two groups,of which one gene was up-regulated and 21 genes were down-regulated;(2)Genes that expressed differently had concentrated functions,biological process involved keratinization,epidermis development and intermediate filament cytoskeleton organization,cellular component related to keratin filament,intermediate filament,extracellular exosome and haptoglobin-hemoglobin complex,molecular function related to structural molecule activity and structural constituent of cytoskeleton;(3)Significantly enriched PANTHER pathways were plasminogen activating cascade,p38 MAPK pathway,oxidative stress response and p53 pathway.Conclusions Trabecular meshwork and extracellular matrix remodeling due to abnormal keratin expression,structural change of intermediate filament cytoskeleton and misregulation of plasminogen activating cascade,p38 MAPK pathway were possible etiology of POAG.Differential expressed genes that related to POAG mainly involve cytoskeleton associated genes

关 键 词：原发性开角型青光眼 小梁网 RNA测序 

分 类 号：R775.2[医药卫生—眼科]