Inhibition of PRKAA/AMPK (Ser485/491) phosphorylation by crizotinib induces cardiotoxicity via perturbing autophagosome-lysosome fusion.
Inhibition of PRKAA/AMPK (Ser485/491) phosphorylation by crizotinib induces cardiotoxicity via perturbing autophagosome-lysosome fusion.
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DOI：
10.1080/15548627.2023.2259216
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发表时间：
2024-02
期刊：
影响因子：
13.3
通讯作者：
Luo, Peihua
中科院分区：
文献类型：
--
作者：
Xu, Zhifei;Pan, Zezheng;Jin, Ying;Gao, Zizheng;Jiang, Feng;Fu, Huangxi;Chen, Xueqin;Zhang, Xiaochen;Yan, Hao;Yang, Xiaochun;Yang, Bo;He, Qiaojun;Luo, Peihua
克唑替尼是一种靶向ALK、MET和ROS 1的小分子酪氨酸激酶抑制剂，是ALK阳性转移性非小细胞肺癌的一线药物，与重度（有时是致死性）心力衰竭病例相关，心力衰竭会增加死亡风险。但其发病机制尚不清楚，导致治疗策略缺乏。我们建立了克唑替尼心脏毒性的体内外模型，发现克唑替尼可引起小鼠左心室功能障碍、心肌损伤和病理性重构，并诱导心肌细胞凋亡和线粒体损伤。此外，我们发现克唑替尼通过中断自噬体-溶酶体融合和MET沉默或重新激活大自噬/自噬流来阻止MET蛋白的降解，从而挽救了克唑替尼引起的心肌细胞死亡和线粒体损伤，这表明自噬活性受损是克唑替尼诱导心脏毒性的关键原因。我们进一步证实，二甲双胍恢复PRKAA/AMPK（Ser 485/491）的磷酸化可重新激活心肌细胞中的自噬流，二甲双胍可挽救克唑替尼诱导的心肌细胞损伤和心脏并发症。总之，我们揭示了克唑替尼诱导的心脏毒性的新机制，其中克唑替尼受损的自噬过程通过抑制MET蛋白的降解而导致心肌细胞死亡和心脏损伤，证明了在药物诱导的心脏毒性中阻碍自噬体-溶酶体融合的新功能，指出了PRKAA磷酸化的重要作用，（Ser 485/491），并证实二甲双胍是克唑替尼诱导的心脏毒性的潜在治疗策略。缩略语和首字母缩略词：AAV病毒：腺相关病毒; ACAC/ACC：乙酰辅酶A羧化酶;放大器：腺苷酸;你好AMP-活化蛋白激酶; ATG5：自噬相关5例; ATG7：自噬相关7例; CHX：环己酰亚胺; CKMB：肌酸激酶心肌带; CQ：氯喹; c-PARP：裂聚（ADP-核糖）聚合酶; DAPI：4 ′ 6-二脒基-2-苯基吲哚; EF：射血分数; FOXO：叉头盒O; FS：缩短分数; GSEA：基因集富集分析; H&E：苏木精和伊红; HF：心力衰竭; HW：TL：心脏重量与胫骨长度的比率; IR：缺血-再灌注; KEGG：基因和基因组的京都百科全书; LAMP 2：溶酶体相关膜蛋白2; LDH：乳酸脱氢酶; MCMs：小鼠心肌细胞;基质金属蛋白酶：线粒体膜电位;线粒体DNA：线粒体DNA; MYH6：肌球蛋白，重肽6，心肌，α; MYH 7：肌球蛋白，重肽7，心肌，β; NPPA：A型利钠肽; NPP B：B型利钠肽; PI：碘化丙啶; PI 3 K：磷酸肌醇3-激酶; PRKAA/AMPKα：蛋白激酶AMP活化催化亚基α; qPCR：定量实时PCR; SD：标准差; SR B：磺酰罗丹明B; TKI：酪氨酸激酶抑制剂; WGA：麦胚凝集素
Crizotinib, a small-molecule tyrosine kinase inhibitor targeting ALK, MET and ROS1, is the first-line drug for ALK-positive metastatic non-small cell lung cancer and is associated with severe, sometimes fatal, cases of cardiac failure, which increases the risk of mortality. However, the underlying mechanism remains unclear, which causes the lack of therapeutic strategy. We established in vitro and in vivo models for crizotinib-induced cardiotoxicity and found that crizotinib caused left ventricular dysfunction, myocardial injury and pathological remodeling in mice and induced cardiomyocyte apoptosis and mitochondrial injury. In addition, we found that crizotinib prevented the degradation of MET protein by interrupting autophagosome-lysosome fusion and silence of MET or re-activating macroautophagy/autophagy flux rescued the cardiomyocytes death and mitochondrial injury caused by crizotinib, suggesting that impaired autophagy activity is the key reason for crizotinib-induced cardiotoxicity. We further confirmed that recovering the phosphorylation of PRKAA/AMPK (Ser485/491) by metformin re-activated autophagy flux in cardiomyocytes and metformin rescued crizotinib-induced cardiomyocyte injury and cardiac complications. In summary, we revealed a novel mechanism for crizotinib-induced cardiotoxicity, wherein the crizotinib-impaired autophagy process causes cardiomyocyte death and cardiac injury by inhibiting the degradation of MET protein, demonstrated a new function of impeded autophagosome-lysosome fusion in drugs-induced cardiotoxicity, pointed out the essential role of the phosphorylation of PRKAA (Ser485/491) in autophagosome-lysosome fusion and confirmed metformin as a potential therapeutic strategy for crizotinib-induced cardiotoxicity. Abbreviations and Acronyms: AAV: adeno-associated virus; ACAC/ACC: acetyl-Co A carboxylase; AMP: adenosine monophosphate; AMPK: AMP-activated protein kinase; ATG5: autophagy related 5; ATG7: autophagy related 7; CHX: cycloheximide; CKMB: creatine kinase myocardial band; CQ: chloroquine; c-PARP: cleaved poly (ADP-ribose) polymerase; DAPI: 4ʹ6-diamidino-2-phenylindole; EF: ejection fraction; FOXO: forkhead box O; FS: fractional shortening; GSEA: gene set enrichment analysis; H&E: hematoxylin and eosin; HF: heart failure; HW: TL: ratio of heart weight to tibia length; IR: ischemia-reperfusion; KEGG: Kyoto encyclopedia of genes and genomes; LAMP2: lysosomal-associated membrane protein 2; LDH: lactate dehydrogenase; MCMs: mouse cardiomyocytes; MMP: mitochondrial membrane potential; mtDNA: mitochondrial DNA; MYH6: myosin, heavy peptide 6, cardiac muscle, alpha; MYH7: myosin, heavy peptide 7, cardiac muscle, beta; NPPA: natriuretic peptide type A; NPPB: natriuretic peptide type B; PI: propidium iodide; PI3K: phosphoinositide 3-kinase; PRKAA/AMPKα: protein kinase AMP-activated catalytic subunit alpha; qPCR: quantitative real-time PCR; SD: standard deviation; SRB: sulforhodamine B; TKI: tyrosine kinase inhibitor; WGA: wheat germ agglutinin
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影响因子：
16.6
作者：
Gélinas R;Mailleux F;Dontaine J;Bultot L;Demeulder B;Ginion A;Daskalopoulos EP;Esfahani H;Dubois-Deruy E;Lauzier B;Gauthier C;Olson AK;Bouchard B;Des Rosiers C;Viollet B;Sakamoto K;Balligand JL;Vanoverschelde JL;Beauloye C;Horman S;Bertrand L
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影响因子：
37.8
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影响因子：
5.6
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