先天性纯红细胞再生障碍性贫血斑马鱼模型相关microRNA的组学研究  

microRNome analysis of the Zebrafish model of diamond-blackfan anemia from Rps19 deficiency

作　　者：张倩[1] 宋斌峰 万扬 张昭军[1] 鲍丙浩 谢尚论 刘汉芝 竺晓凡 袁卫平 贾海波[2] 方向东[1] 

机构地区：[1]中国科学院北京基因组研究所中科院基因组学与信息重点实验室,北京100101 [2]华中科技大学生命科学与技术学院人类基因组研究中心分子生物物理教育部重点实验室,武汉430074 [3]中国医学科学院协和医学院血液病医院血液学研究所,实验血液学国家重点实验室,天津300020

出　　处：《发育医学电子杂志》2014年第3期139-145,共7页Journal of Developmental Medicine (Electronic Version)

基　　金：国家"十二五"科技支撑计划分课题(2013BAI01B09);国家重大科学仪器设备开发专项子课题(2011YQ03013404);国家自然科学基金(31171387和81090414)

摘　　要：目的探讨先天性纯红细胞再生障碍性贫血(DBA)相关microRNA的表达特征及其调控机制。方法利用morpholino技术建立核糖体蛋白基因Rps19表达不足的DBA斑马鱼模型和Rps19与p53同时表达不足的斑马鱼模型,利用新一代高通量测序(RNA-seq)技术,研究DBA斑马鱼模型的microRNA表达谱的特征及p53在其中的调控作用。结果 Rps19 MO的斑马鱼胚胎的循环血细胞明显减少,且尾部呈现出向腹侧弯曲的异常形态。而在p53敲低之后,斑马鱼的形态异常得到了一定程度的恢复。通过对microRNA表达谱的比较分析发现,在Rps19不足的斑马鱼胚胎中约1/4的microRNA表达量显著偏低,这些异常表达的microRNA可能影响到斑马鱼的正常发育过程。而在p53敲低之后,部分microRNA的表达恢复到正常水平。最终确定了47个在DBA发病过程中可能受p53调控的microRNA。结论 DBA斑马鱼模型的microRNA普遍低表达,其中部分microRNA受到p53的表达水平的调控。Objective To characterize the genome-wide microRNA profile and to explore the biological functions and probable mechanisms for microRNAs in diamond-blackfan anemia (DBA). Method Zebrafish embryos at the one-cell stage were injected with the Rps19 MO and Rps19+p53 MO. The miRNomes of these zebrafish models and control, generated by miRNA-seq, were compared for determining the microRNA profile characteristics of DBA and regulatory function of p53. Result Rps19 morphants showed an obviously ventrally bent tail and a reduction in the circulating blood cells compared with the control embryos, which were partially rescued in Rps19+p53 MO at 48 hpf. By comparing the miRNomes with control, we found that about 25% of total expressed microRNAs in Rps19 MO had significantly lower expression, which might make an influence on the development process of Zebrafish embryos. The abnormal miRNomes were partially reversed by simultaneous knockdown of p53. We determined 47 microRNAs that were potentially regulated by p53 in DBA pathologic process.Conclusion microRNAs in Zebrafish model of DBA were in an inactive state, which was partially affected by p53.

关 键 词：纯红细胞再生障碍性贫血 MICRORNA Rps19 P53 RNA-SEQ 斑马鱼 

分 类 号：R556.5[医药卫生—血液循环系统疾病] R-332[医药卫生—内科学]