Cardiovascular Research Center (Y.H., E.Y., M.M.Ż., E.C., N.S., M.T.K.S., R.K., A.A.K., K.K., A.M., N.H., L.Z., A.F, M.G.K.), Icahn School of Medicine at Mount Sinai, New York.
Department of Genetics and Genomic Sciences (Y.H., E.Y., M.M.Ż., E.C., N.S., M.T.K.S., R.K., A.A.K., K.K., A.M., N.H., L.Z., A.S.V., N.G., E.K., E.S., E.E.), Icahn School of Medicine at Mount Sinai, New York.
Circulation. 2020 Mar 17;141(11):916-930. doi: 10.1161/CIRCULATIONAHA.119.041882. Epub 2020 Jan 29.
Sphingolipids have recently emerged as a biomarker of recurrence and mortality after myocardial infarction (MI). The increased ceramide levels in mammalian heart tissues during acute MI, as demonstrated by several groups, is associated with higher cell death rates in the left ventricle and deteriorated cardiac function. Ceramidase, the only enzyme known to hydrolyze proapoptotic ceramide, generates sphingosine, which is then phosphorylated by sphingosine kinase to produce the prosurvival molecule sphingosine-1-phosphate. We hypothesized that Acid Ceramidase (AC) overexpression would counteract the negative effects of elevated ceramide and promote cell survival, thereby providing cardioprotection after MI.
We performed transcriptomic, sphingolipid, and protein analyses to evaluate sphingolipid metabolism and signaling post-MI. We investigated the effect of altering ceramide metabolism through a loss (chemical inhibitors) or gain (modified mRNA [modRNA]) of AC function post hypoxia or MI.
We found that several genes involved in de novo ceramide synthesis were upregulated and that ceramide (C16, C20, C20:1, and C24) levels had significantly increased 24 hours after MI. AC inhibition after hypoxia or MI resulted in reduced AC activity and increased cell death. By contrast, enhancing AC activity via AC modRNA treatment increased cell survival after hypoxia or MI. AC modRNA-treated mice had significantly better heart function, longer survival, and smaller scar size than control mice 28 days post-MI. We attributed the improvement in heart function post-MI after AC modRNA delivery to decreased ceramide levels, lower cell death rates, and changes in the composition of the immune cell population in the left ventricle manifested by lowered abundance of proinflammatory detrimental neutrophils.
Our findings suggest that transiently altering sphingolipid metabolism through AC overexpression is sufficient and necessary to induce cardioprotection post-MI, thereby highlighting the therapeutic potential of AC modRNA in ischemic heart disease.
鞘脂最近被认为是心肌梗死后复发和死亡的生物标志物。几个研究小组证明,急性心肌梗死后哺乳动物心脏组织中神经酰胺水平升高与左心室细胞死亡率升高和心脏功能恶化有关。神经酰胺酶是唯一已知能够水解促凋亡神经酰胺的酶,它产生神经酰胺,然后被神经酰胺激酶磷酸化生成具有促生存作用的分子神经酰胺-1-磷酸。我们假设酸性神经酰胺酶(AC)过表达将抵消神经酰胺升高的负面影响并促进细胞存活,从而在心肌梗死后提供心脏保护。
我们进行了转录组、鞘脂和蛋白质分析,以评估心肌梗死后的鞘脂代谢和信号转导。我们通过改变 AC 功能(化学抑制剂或修饰的 mRNA [modRNA])来研究改变神经酰胺代谢对缺氧或心肌梗死后的影响。
我们发现,几个参与从头合成神经酰胺的基因上调,并且神经酰胺(C16、C20、C20:1 和 C24)水平在心肌梗死后 24 小时显著升高。缺氧或心肌梗死后 AC 抑制导致 AC 活性降低和细胞死亡增加。相比之下,通过 AC modRNA 处理增强 AC 活性可增加缺氧或心肌梗死后的细胞存活。与对照组相比,AC modRNA 治疗的小鼠在心肌梗死后 28 天具有更好的心脏功能、更长的存活时间和更小的疤痕面积。我们将心肌梗死后心脏功能的改善归因于 AC modRNA 给药后神经酰胺水平降低、细胞死亡率降低以及左心室中免疫细胞群体组成的变化,表现为促炎有害中性粒细胞的丰度降低。
我们的发现表明,通过 AC 过表达短暂改变鞘脂代谢足以诱导心肌梗死后的心脏保护,从而突出了 AC modRNA 在缺血性心脏病中的治疗潜力。