Toronto General Research Institute, 100 College St., M5G 1L7, Toronto, ON, Canada.
Division of Cardiology, University Health Network (UHN), 200 Elizabeth St., Toronto, ON, M5G 2C4, Canada.
Cell Death Differ. 2021 Apr;28(4):1398-1417. doi: 10.1038/s41418-020-00669-9. Epub 2020 Dec 7.
Adult mammalian cardiomyocytes (CM) are postmitotic, differentiated cells that cannot re-enter the cell cycle after any appreciable injury. Therefore, understanding the factors required to induce CM proliferation for repair is of great clinical importance. While expression of muscle pyruvate kinase 2 (Pkm2), a cytosolic enzyme catalyzing the final step in glycolysis, is high in end-stage heart failure (HF), the loss of Pkm2 promotes proliferation in some cellular systems, in vivo. We hypothesized that in the adult heart CM proliferation may require low Pkm2 activity. Thus, we investigated the potential for Pkm2 to regulate CM proliferation in a mouse model of myocardial infarction (MI) employing inducible, cardiac-specific Pkm2 gene knockout (Pkm2KOi) mice. We found a lack of cardiac hypertrophy or expression of the fetal gene program in Pkm2KOi mice post MI, as compared to vehicle control animals (P < 0.01), correlating with smaller infarct size, improved mitochondrial (mt) function, enhanced angiogenesis, reduced degree of CM apoptosis, and reduced oxidative stress post MI. There was significantly higher numbers of dividing CM in the infarct zone between 3-9 days post MI (P < 0.001). Mechanistically, we determined that Pkm2 interacts with β-catenin (Ctnnb1) in the cytoplasm of CM, inhibiting Ctnnb1 phosphorylation at serine 552 and tyrosine 333, by Akt. In the absence of Pkm2, Ctnnb1 translocates to the nucleus leading to transcriptional activation of proliferation-associated target genes. All these effects are abrogated by genetic co-deletion of Pkm2 and Ctnnb1. Collectively, this work supports a novel antiproliferative function for Pkm2 in CM through the sequestration of Ctnnb1 in the cytoplasm of CM whereas loss of Pkm2 is essential for CM proliferation. Reducing cardiac Pkm2 expression may provide a useful strategy for cardiac repair after MI in patients.
成年哺乳动物心肌细胞(CM)是有丝分裂后分化的细胞,在受到任何明显损伤后都无法重新进入细胞周期。因此,了解诱导 CM 增殖以进行修复所需的因素具有重要的临床意义。虽然肌肉丙酮酸激酶 2(Pkm2)的表达在终末期心力衰竭(HF)中很高,这种催化糖酵解最后一步的胞质酶,但在一些细胞系统中,Pkm2 的缺失会促进增殖,在体内也是如此。我们假设在成年心脏中,CM 的增殖可能需要低 Pkm2 活性。因此,我们使用诱导型心脏特异性 Pkm2 基因敲除(Pkm2KOi)小鼠模型,研究了 Pkm2 调节 CM 增殖的潜力。我们发现,与 vehicle 对照动物(P < 0.01)相比,Pkm2KOi 小鼠在 MI 后缺乏心肌肥厚或胎儿基因程序的表达,这与梗死面积较小、线粒体(mt)功能增强、血管生成增加、CM 凋亡减少和氧化应激减少相关。在 MI 后 3-9 天,梗死区的分裂 CM 数量明显增多(P < 0.001)。从机制上讲,我们确定 Pkm2 在 CM 的细胞质中与β-连环蛋白(Ctnnb1)相互作用,通过 Akt 抑制 Ctnnb1 丝氨酸 552 和酪氨酸 333 的磷酸化。在缺乏 Pkm2 的情况下,Ctnnb1 易位到细胞核,导致增殖相关靶基因的转录激活。所有这些效应都被 Pkm2 和 Ctnnb1 的遗传共缺失所消除。总的来说,这项工作支持了 Pkm2 在 CM 中的一种新的抗增殖功能,即通过将 Ctnnb1 隔离在 CM 的细胞质中来实现,而 Pkm2 的缺失对于 CM 的增殖是必不可少的。降低心脏 Pkm2 的表达可能为 MI 后患者的心脏修复提供一种有用的策略。