Phosphoproteomic Analysis of Neonatal Regenerative Myocardium Revealed Important Roles of Checkpoint Kinase 1 via Activating Mammalian Target of Rapamycin C1/Ribosomal Protein S6 Kinase b-1 Pathway
Phosphoproteomic Analysis of Neonatal Regenerative Myocardium Revealed Important Roles of Checkpoint Kinase 1 via Activating Mammalian Target of Rapamycin C1/Ribosomal Protein S6 Kinase b-1 Pathway
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新生儿再生心肌的磷酸化蛋白质组学分析揭示了检查点激酶 1 通过激活雷帕霉素 C1/核糖体蛋白 S6 激酶 b-1 通路的哺乳动物靶点的重要作用
DOI：
10.1161/circulationaha.119.040747
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发表时间：
2020-05-12
期刊：
影响因子：
37.8
通讯作者：
Wang, Liansheng
中科院分区：
文献类型：
--
作者：
Fan, Yi;Cheng, Yiwei;Wang, Liansheng
背景：在哺乳动物中，心肌梗塞后的再生治疗受到成人心脏有限的再生能力的阻碍，而新生儿心脏则维持短暂的再生能力。对缺血新生儿心肌的系统磷酸化信号分析可能有助于确定参与心脏再生的关键途径。我们的目的是定义缺血性新生儿心肌中的激酶-底物网络，并确定缺血性损伤后参与心脏再生的关键通路。方法：对新生儿心肌梗死边界区进行定量磷酸化蛋白质组学分析，激酶-底物网络分析显示 11 种底物丰富且磷酸化水平上调的激酶，包括检查点激酶 1 (CHK1) 激酶。在接受心尖切除或心肌梗塞的癌症研究所 CD1 新生和成年小鼠身上测试了 CHK1 对心脏再生的影响。结果：在体外，CHK1 过度表达促进心肌细胞增殖，而 CHK1 敲除则减弱心肌细胞增殖。在体内，抑制 CHK1 会阻碍新生小鼠切除边界区的心肌再生。在成年心肌梗死小鼠中，梗死边界区CHK1过表达上调雷帕霉素C1/核糖体蛋白S6激酶b-1通路的哺乳动物靶点，促进心肌细胞增殖，改善心功能。雷帕霉素抑制哺乳动物雷帕霉素靶点活性，可抑制体外 CHK1 过表达诱导的新生儿心肌细胞增殖。结论：我们的研究表明，新生儿再生心肌的磷酸化蛋白质组有助于识别参与心肌再生的重要信号通路。 CHK1被发现是负责新生儿再生的关键信号。 CHK1 的心肌过度表达可以通过激活雷帕霉素 C1/核糖体蛋白 S6 激酶 b-1 通路的哺乳动物靶点来改善成人心脏的心脏再生。因此，CHK1可能作为心肌梗死后心肌修复的潜在新靶点。
BACKGROUND: In mammals, regenerative therapy after myocardial infarction is hampered by the limited regenerative capacity of adult heart, whereas a transient regenerative capacity is maintained in the neonatal heart. Systemic phosphorylation signaling analysis on ischemic neonatal myocardium might be helpful to identify key pathways involved in heart regeneration. Our aim was to define the kinase-substrate network in ischemic neonatal myocardium and to identify key pathways involved in heart regeneration after ischemic insult.METHODS: Quantitative phosphoproteomics profiling was performed on infarct border zone of neonatal myocardium, and kinase-substrate network analysis revealed 11 kinases with enriched substrates and upregulated phosphorylation levels, including checkpoint kinase 1 (CHK1) kinase. The effect of CHK1 on cardiac regeneration was tested on Institute of Cancer Research CD1 neonatal and adult mice that underwent apical resection or myocardial infarction.RESULTS: In vitro, CHK1 overexpression promoted whereas CHK1 knockdown blunted cardiomyocyte proliferation. In vivo, inhibition of CHK1 hindered myocardial regeneration on resection border zone in neonatal mice. In adult myocardial infarction mice, CHK1 overexpression on infarct border zone upregulated mammalian target of rapamycin C1/ribosomal protein S6 kinase b-1 pathway, promoted cardiomyocyte proliferation, and improved cardiac function. Inhibiting mammalian target of rapamycin activity by rapamycin blunted the neonatal cardiomyocyte proliferation induced by CHK1 overexpression in vitro.CONCLUSIONS: Our study indicates that phosphoproteome of neonatal regenerative myocardium could help identify important signaling pathways involved in myocardial regeneration. CHK1 is found to be a key signaling responsible for neonatal regeneration. Myocardial overexpression of CHK1 could improve cardiac regeneration in adult hearts by activating the mammalian target of rapamycin C1/ribosomal protein S6 kinase b-1 pathway. Thus, CHK1 might serve as a potential novel target in myocardial repair after myocardial infarction.