Circulating markers of NADH-reductive stress correlate with mitochondrial disease severity
Circulating markers of NADH-reductive stress correlate with mitochondrial disease severity
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DOI：
10.1172/jci136055
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发表时间：
2021-01-19
影响因子：
15.9
通讯作者：
Mootha, Vamsi K.
中科院分区：
文献类型：
--
作者：
Sharma, Rohit;Reinstadler, Bryn;Mootha, Vamsi K.
线粒体疾病代表了大量罕见的综合征，这些综合征很难管理，因为我们不完全了解生化发病机制，而且我们目前缺乏简单的严重程度标志物。m.3243A>G变异是最常见的异质性线粒体DNA突变，并且是一系列疾病的基础，特别是线粒体脑肌病乳酸酸中毒和中风样发作（MELAS）。为了鉴定m.3243A>G疾病的稳健循环标志物，我们首先对来自深度表型化队列（102名患者，32名对照）的血浆进行发现蛋白质组学、靶向代谢组学和非靶向代谢组学。在验证阶段，我们使用不同的方法测量了独立队列中优先代谢物的浓度。我们验证了20种分析物（1种蛋白质，19种代谢物），可区分MELAS患者和对照组。收集包括经典的（乳酸，丙氨酸）和最近确定的（GDF-15，α-羟基丁酸）线粒体标记物。通过挖掘非靶向质谱，我们发现了3个研究较少的代谢产物家族：N-乳酰氨基酸，β-羟基酰基肉毒碱和β-羟基脂肪酸。这20种分析物中的许多与已建立的严重性测量（包括Karnofsky状态）密切相关，并且从机制上讲，几乎所有标记物都可归因于升高的NADH/NAD（+）比率或NADH还原应激。我们的工作定义了一组与NADH还原应激相关的细胞器功能测试，这些测试应该能够对线粒体疾病进行分类和监测。
Mitochondrial disorders represent a large collection of rare syndromes that are difficult to manage both because we do not fully understand biochemical pathogenesis and because we currently lack facile markers of severity. The m.3243A>G variant is the most common heteroplasmic mitochondrial DNA mutation and underlies a spectrum of diseases, notably mitochondrial encephalomyopathy lactic acidosis and stroke-like episodes (MELAS). To identify robust circulating markers of m.3243A>G disease, we first performed discovery proteomics, targeted metabolomics, and untargeted metabolomics on plasma from a deeply phenotyped cohort (102 patients, 32 controls). In a validation phase, we measured concentrations of prioritized metabolites in an independent cohort using distinct methods. We validated 20 analytes (1 protein, 19 metabolites) that distinguish patients with MELAS from controls. The collection includes classic (lactate, alanine) and more recently identified (GDF-15, alpha-hydroxybutyrate) mitochondrial markers. By mining untargeted mass-spectra we uncovered 3 less well-studied metabolite families: N-lactoyl-amino acids, beta-hydroxy acylcarnitines, and beta-hydroxy fatty acids. Many of these 20 analytes correlate strongly with established measures of severity, including Karnofsky status, and mechanistically, nearly all markers are attributable to an elevated NADH/NAD(+) ratio, or NADH-reductive stress. Our work defines a panel of organelle function tests related to NADH-reductive stress that should enable classification and monitoring of mitochondrial disease.