The Tumor-Suppressive Human Circular RNA CircITCH Sponges miR-330-5p to Ameliorate Doxorubicin-Induced Cardiotoxicity Through Upregulating SIRT6, Survivin, and SERCA2a
The Tumor-Suppressive Human Circular RNA CircITCH Sponges miR-330-5p to Ameliorate Doxorubicin-Induced Cardiotoxicity Through Upregulating SIRT6, Survivin, and SERCA2a
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肿瘤抑制性人类环状 RNA CircITCH 海绵状 miR-330-5p 通过上调 SIRT6、Survivin 和 SERCA2a 改善多柔比星诱导的心脏毒性
DOI：
10.1161/circresaha.119.316061
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发表时间：
2020-07-31
影响因子：
20.1
通讯作者：
Cao, Feng
中科院分区：
文献类型：
--
作者：
Han, Dong;Wang, Yongjun;Cao, Feng
补充数字内容可在文本中找到。理由：阿霉素是最有效的抗肿瘤药物之一；然而，它的临床使用受到限制，因为它有严重的心脏毒性风险。先前的研究已经证实，CircITCH （circular RNA ITCH [E3泛素蛋白连接酶]）是一种广谱的肿瘤抑制环状RNA，其宿主基因ITCH （E3泛素蛋白连接酶）参与了阿霉素诱导的心脏毒性（DOXIC）。CircITCH是否在DOXIC中起作用尚不清楚。目的：探讨CircITCH在DOXIC中的作用，并进一步揭示其潜在机制。方法和结果：环状RNA测序筛选可能参与DOXI发病机制的环状RNA。定量聚合酶链反应和RNA原位杂交显示，CircITCH在阿霉素处理的人诱导的多能干细胞来源的心肌细胞（hiPSC-CMs）以及患有阿霉素诱导的心肌病的癌症患者的尸检标本中下调。细胞死亡/活力测定、心肌细胞坏死标志物检测、微电极阵列和心肌细胞功能测定显示，CircITCH改善了阿霉素诱导的心肌细胞损伤和功能障碍。细胞/线粒体氧化应激和DNA损伤标志物检测证实CircITCH减轻了阿霉素诱导的细胞/线粒体氧化应激和DNA损伤。RNA下拉实验、Ago2免疫沉淀和双荧光原位杂交鉴定miR-330-5p是CircITCH的直接靶点。此外，CircITCH被发现通过作为内源性海绵隔离miR-330-5p发挥作用。生物信息学分析、荧光素酶报告基因测定和定量聚合酶链反应表明SIRT6 （sirtuin 6）、BIRC5 （baculloviral IAP repeat containing 5, Survivin）和ATP2A2 （atp酶肌浆/内质网Ca2+转运2，SERCA2a [SR Ca2+- atp酶2]）是miR-330-5p的直接靶标，它们在DOXIC中受CircITCH/miR-330-5p轴的调控。进一步的实验表明，circitch介导的DOXIC减轻依赖于miR-330-5p与SIRT6、BIRC5和ATP2A2 mRNA的3 ' - utr之间的相互作用。最后，基于AAV9（腺相关病毒血清型9）载体过表达保守的CircITCH部分阻止了小鼠DOXIC。结论：CircITCH是DOXIC的一个新的治疗靶点，因为它作为miR-330-5p的天然海绵，从而上调SIRT6、Survivin和SERCA2a，以减轻阿霉素诱导的心肌细胞损伤和功能障碍。
Supplemental Digital Content is available in the text. Rationale: Doxorubicin is one of the most potent antitumor agents available; however, its clinical use is restricted because it poses a risk of severe cardiotoxicity. Previous work has established that CircITCH (circular RNA ITCH [E3 ubiquitin-protein ligase]) is a broad-spectrum tumor-suppressive circular RNA and that its host gene, ITCH (E3 ubiquitin protein ligase), is involved in doxorubicin-induced cardiotoxicity (DOXIC). Whether CircITCH plays a role in DOXIC remains unknown. Objective: We aimed to dissect the role of CircITCH in DOXIC and further decipher its potential mechanisms. Methods and Results: Circular RNA sequencing was performed to screen the potentially involved circRNAs in DOXI pathogenesis. Quantitative polymerase chain reaction and RNA in situ hybridization revealed that CircITCH was downregulated in doxorubicin-treated human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) as well as in the autopsy specimens from cancer patients who suffered from doxorubicin-induced cardiomyopathy. Cell death/viability assays, detection of cardiomyocyte necrosis markers, microelectrode array, and cardiomyocyte functional assays revealed that CircITCH ameliorated doxorubicin-induced cardiomyocyte injury and dysfunction. Detection of cellular/mitochondrial oxidative stress and DNA damage markers verified that CircITCH alleviated cellular/mitochondrial oxidative stress and DNA damage induced by doxorubicin. RNA pull-down assays, Ago2 immunoprecipitation and double fluorescent in situ hybridization identified miR-330-5p as a direct target of CircITCH. Moreover, CircITCH was found to function by acting as an endogenous sponge that sequestered miR-330-5p. Bioinformatic analysis, luciferase reporter assays, and quantitative polymerase chain reaction showed that SIRT6 (sirtuin 6), BIRC5 (baculoviral IAP repeat containing 5, Survivin), and ATP2A2 (ATPase sarcoplasmic/endoplasmic reticulum Ca2+ transporting 2, SERCA2a [SR Ca2+-ATPase 2]) were direct targets of miR-330-5p and that they were regulated by the CircITCH/miR-330-5p axis in DOXIC. Further experiments demonstrated that CircITCH-mediated alleviation of DOXIC was dependent on the interactions between miR-330-5p and the 3′-UTRs of SIRT6, BIRC5, and ATP2A2 mRNA. Finally, AAV9 (adeno-associated virus serotype 9) vector-based overexpression of the well-conserved CircITCH partly prevented DOXIC in mice. Conclusions: CircITCH represents a novel therapeutic target for DOXIC because it acts as a natural sponge of miR-330-5p, thereby upregulating SIRT6, Survivin and SERCA2a to alleviate doxorubicin-induced cardiomyocyte injury and dysfunction.