Inhibition of the Hypoxia-Inducible Factor 1α-Induced Cardiospecific HERNA1 Enhance-Templated RNA Protects From Heart Disease
Inhibition of the Hypoxia-Inducible Factor 1α-Induced Cardiospecific HERNA1 Enhance-Templated RNA Protects From Heart Disease
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DOI：
10.1161/circulationaha.118.036769
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发表时间：
2019-06-11
期刊：
影响因子：
37.8
通讯作者：
Krishnan, Jaya
中科院分区：
文献类型：
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作者：
Mirtschink, Peter;Bischof, Corinne;Krishnan, Jaya
背景：增强子是基因组调控元件，赋予基因表达的时空和信号依赖性控制。最近的证据表明，增强子可以产生非编码增强子RNA，但它们的（病理）生物学功能在很大程度上仍然难以捉摸。我们进行了组蛋白标记的染色质免疫沉淀偶联测序，并结合来自人类心脏肥大实验和遗传小鼠模型的左心室活检的RNA测序，以鉴定揭示增强子定位、与人类基因组的保守性和缺氧诱导因子1a依赖性。最有前途的候选者，缺氧诱导增强子RNA（HERNA）1，通过研究其调节相邻的编码基因表达的能力，通过使用染色质分离RNA纯化和N-BoxB tethering-based报告基因测定结合到他们的基因启动子进行进一步检查。HERNA 1及其邻近基因对病理性应激诱导的生长和收缩功能障碍的作用，以及HERNA 1抑制的治疗潜力在gapmer介导的功能丧失研究中进行了研究，在体外使用人诱导多能干细胞衍生的心肌细胞和各种人病理性心脏肥大的体内模型。HERNA 1的产生是通过直接缺氧诱导因子1a与增强子的组蛋白H3-赖氨酸27乙酰化标记富集启动子中的缺氧反应元件结合而启动的，并将缺氧反应性赋予附近的基因，包括突触结合蛋白XVII（膜运输和Ca 2+感应蛋白家族的成员）和SMG 1（编码磷脂酰肌醇3-激酶相关激酶）。因此，SMG 1的底物ATP依赖性RNA解旋酶上移码1以HERNA 1和SMG 1依赖性方式被过度磷酸化。SMG 1和SYT 17的体外和体内失活揭示了在调节心脏肥大中重叠和不同的作用。最后，体内施用靶向HERNA 1的反义寡核苷酸保护小鼠免受应激诱导的病理性肥大。抑制HERNA 1 postdisease的发展逆转左心室的生长和功能障碍，导致增加的总survival.CONCLUSIONS：HERNA 1是一种新的心脏特异性非编码RNA的关键调控功能，在调节疾病的生长，代谢和收缩基因程序，并揭示了一个分子靶点，适合治疗开发。
BACKGROUND: Enhancers are genomic regulatory elements conferring spatiotemporal and signal-dependent control of gene expression. Recent evidence suggests that enhancers can generate noncoding enhancer RNAs, but their (patho) biological functions remain largely elusive.METHODS: We performed chromatin immunoprecipitation-coupled sequencing of histone marks combined with RNA sequencing of left ventricular biopsies from experimental and genetic mouse models of human cardiac hypertrophy to identify transcripts revealing enhancer localization, conservation with the human genome, and hypoxia-inducible factor 1a dependence. The most promising candidate, hypoxia-inducible enhancer RNA (HERNA) 1, was further examined by investigating its capacity to modulate neighboring coding gene expression by binding to their gene promoters by using chromatin isolation by RNA purification and.N-BoxB tetheringbased reporter assays. The role of HERNA1 and its neighboring genes for pathological stress-induced growth and contractile dysfunction, and the therapeutic potential of HERNA1 inhibition was studied in gapmer-mediated loss-of-function studies in vitro using human induced pluripotent stem cellderived cardiomyocytes and various in vivo models of human pathological cardiac hypertrophy.RESULTS: HERNA1 is robustly induced on pathological stress. Production of HERNA1 is initiated by direct hypoxia-inducible factor 1a binding to a hypoxiaresponse element in the histoneH3-lysine27acetylation marks-enriched promoter of the enhancer and confers hypoxia responsiveness to nearby genes including synaptotagmin XVII, a member of the family of membrane-trafficking and Ca2+-sensing proteins and SMG1, encoding a phosphatidylinositol 3-kinase-related kinase. Consequently, a substrate of SMG1, ATP-dependent RNA helicase upframeshift 1, is hyperphoshorylated in a HERNA1-and SMG1dependent manner. In vitro and in vivo inactivation of SMG1 and SYT17 revealed overlapping and distinct roles in modulating cardiac hypertrophy. Finally, in vivo administration of antisense oligonucleotides targeting HERNA1 protected mice from stress-induced pathological hypertrophy. The inhibition of HERNA1 postdisease development reversed left ventricular growth and dysfunction, resulting in increased overall survival.CONCLUSIONS: HERNA1 is a novel heart-specific noncoding RNA with key regulatory functions in modulating the growth, metabolic, and contractile gene program in disease, and reveals a molecular target amenable to therapeutic exploitation.