The beagle dog MicroRNA tissue atlas: identifying translatable biomarkers of organ toxicity.
The beagle dog MicroRNA tissue atlas: identifying translatable biomarkers of organ toxicity.
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DOI：
10.1186/s12864-016-2958-x
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发表时间：
2016-08-17
期刊：
影响因子：
4.4
通讯作者：
Kirby P
中科院分区：
文献类型：
--
作者：
Koenig EM;Fisher C;Bernard H;Wolenski FS;Gerrein J;Carsillo M;Gallacher M;Tse A;Peters R;Smith A;Meehan A;Tirrell S;Kirby P
MicroRNA（miRNA）是一类长度在25个核苷酸以下的单链非编码RNA，通过翻译抑制或mRNA降解来调节转录后基因的表达。在组织损伤后的体循环中可以检测到升高水平的miRNA，这表明miRNA在细胞损伤后释放。由于其显著的稳定性，易于在生物流体中检测，以及组织特异性表达模式，miRNA有可能成为器官损伤的特异性生物标志物。miRNA生物标志物的鉴定需要系统的方法：1）通过下一代测序确定哺乳动物物种内的miRNA组织表达谱; 2）鉴定毒理学感兴趣的器官内富集的和/或特异性的miRNA表达，以及3）用组织特异性毒物进行体内验证。虽然在啮齿类动物和人类中已经报道了miRNA组织表达，但在狗（一种相关毒理学种属）中的miRNA组织表达数据很少。这里描述了第一个狗miRNA组织图谱的生成和评估。对来自5只雄性比格犬的16个组织的分析鉴定了106个组织富集的miRNA，其中60个在单个器官中高度富集，因此可以作为器官损伤的生物标志物。在给予肝毒性物质的犬中进行的概念验证研究评价了对肝脏、心脏、骨骼肌、胰腺、睾丸和脑特异性的15种组织富集miRNA的qPCR组。血清中miR-122和miR-885水平升高的犬丙氨酸氨基转移酶也相应升高，显微镜分析证实了肝损伤。筛选组中包括的其他非肝脏富集的miRNA不受影响。Eli Lilly作者创建了一个免费的Sprague Dawely大鼠miRNA组织图谱，并证明了在大鼠和犬中给予雨蛙肽后，循环中胰腺富集的miRNA水平增加。狗的miRNA组织图谱提供了生物标志物发现的资源，并且可以通过狗基因组注释的细化来进一步挖掘。在狗miRNA组织图谱中鉴定的60种高度富集的组织miRNA可以作为诊断生物标志物，并且需要通过与组织病理学的体内相关性进行进一步验证。一旦得到验证，这些组织富集的miRNAs可以组合成一个强大的qPCR筛选小组，以在早期药物开发期间鉴定器官毒性。本文的在线版本（doi：10.1186/s12864-016-2958-x）包含补充材料，可供授权用户使用。
MicroRNAs (miRNA) are varied in length, under 25 nucleotides, single-stranded noncoding RNA that regulate post-transcriptional gene expression via translational repression or mRNA degradation. Elevated levels of miRNAs can be detected in systemic circulation after tissue injury, suggesting that miRNAs are released following cellular damage. Because of their remarkable stability, ease of detection in biofluids, and tissue specific expression patterns, miRNAs have the potential to be specific biomarkers of organ injury. The identification of miRNA biomarkers requires a systematic approach: 1) determine the miRNA tissue expression profiles within a mammalian species via next generation sequencing; 2) identify enriched and/or specific miRNA expression within organs of toxicologic interest, and 3) in vivo validation with tissue-specific toxicants. While miRNA tissue expression has been reported in rodents and humans, little data exists on miRNA tissue expression in the dog, a relevant toxicology species. The generation and evaluation of the first dog miRNA tissue atlas is described here. Analysis of 16 tissues from five male beagle dogs identified 106 tissue enriched miRNAs, 60 of which were highly enriched in a single organ, and thus may serve as biomarkers of organ injury. A proof of concept study in dogs dosed with hepatotoxicants evaluated a qPCR panel of 15 tissue enriched miRNAs specific to liver, heart, skeletal muscle, pancreas, testes, and brain. Dogs with elevated serum levels of miR-122 and miR-885 had a correlative increase of alanine aminotransferase, and microscopic analysis confirmed liver damage. Other non-liver enriched miRNAs included in the screening panel were unaffected. Eli Lilly authors created a complimentary Sprague Dawely rat miRNA tissue atlas and demonstrated increased pancreas enriched miRNA levels in circulation, following caerulein administration in rat and dog. The dog miRNA tissue atlas provides a resource for biomarker discovery and can be further mined with refinement of dog genome annotation. The 60 highly enriched tissue miRNAs identified within the dog miRNA tissue atlas could serve as diagnostic biomarkers and will require further validation by in vivo correlation to histopathology. Once validated, these tissue enriched miRNAs could be combined into a powerful qPCR screening panel to identify organ toxicity during early drug development. The online version of this article (doi:10.1186/s12864-016-2958-x) contains supplementary material, which is available to authorized users.