Jung SeungWoo, Bohan Amy
Am J Vet Res. 2018 Feb;79(2):163-169. doi: 10.2460/ajvr.79.2.163.
OBJECTIVE To characterize expression profiles of circulating microRNAs via genome-wide sequencing for dogs with congestive heart failure (CHF) secondary to myxomatous mitral valve degeneration (MMVD). ANIMALS 9 healthy client-owned dogs and 8 age-matched client-owned dogs with CHF secondary to MMVD. PROCEDURES Blood samples were collected before administering cardiac medications for the management of CHF. Isolated microRNAs from plasma were classified into microRNA libraries and subjected to next-generation sequencing (NGS) for genome-wide sequencing analysis and quantification of circulating microRNAs. Quantitative reverse transcription PCR (qRT-PCR) assays were used to validate expression profiles of differentially expressed circulating microRNAs identified from NGS analysis of dogs with CHF. RESULTS 326 microRNAs were identified with NGS analysis. Hierarchical analysis revealed distinct expression patterns of circulating microRNAs between healthy dogs and dogs with CHF. Results of qRT-PCR assays confirmed upregulation of 4 microRNAs (miR-133, miR-1, miR-let-7e, and miR-125) and downregulation of 4 selected microRNAs (miR-30c, miR-128, miR-142, and miR-423). Results of qRT-PCR assays were highly correlated with NGS data and supported the specificity of circulating microRNA expression profiles in dogs with CHF secondary to MMVD. CONCLUSIONS AND CLINICAL RELEVANCE These results suggested that circulating microRNA expression patterns were unique and could serve as molecular biomarkers of CHF in dogs with MMVD.
目的 通过全基因组测序来表征患有黏液瘤性二尖瓣退化(MMVD)继发充血性心力衰竭(CHF)的犬循环微小RNA的表达谱。动物 9只健康的客户拥有的犬和8只年龄匹配的患有MMVD继发CHF的客户拥有的犬。程序 在给予治疗CHF的心脏药物之前采集血样。从血浆中分离的微小RNA被分类到微小RNA文库中,并进行下一代测序(NGS)以进行全基因组测序分析和循环微小RNA的定量。定量逆转录PCR(qRT-PCR)测定用于验证从CHF犬的NGS分析中鉴定出的差异表达的循环微小RNA的表达谱。结果 NGS分析鉴定出326种微小RNA。层次分析揭示了健康犬和CHF犬之间循环微小RNA的不同表达模式。qRT-PCR测定结果证实4种微小RNA(miR-133、miR-1、miR-let-7e和miR-125)上调,4种选定的微小RNA(miR-30c、miR-128、miR-142和miR-423)下调。qRT-PCR测定结果与NGS数据高度相关,并支持MMVD继发CHF犬循环微小RNA表达谱的特异性。结论及临床意义 这些结果表明,循环微小RNA表达模式是独特的,可作为MMVD继发CHF犬CHF的分子生物标志物。