Mitochondrial GSNOR Alleviates Cardiac Dysfunction via ANT1 Denitrosylation
Mitochondrial GSNOR Alleviates Cardiac Dysfunction via ANT1 Denitrosylation
复制标题
DOI：
10.1161/circresaha.123.322654
复制
发表时间：
2023-06
影响因子：
20.1
通讯作者：
Xin Tang;Shuang Zhao;Jieqiong Liu;Xiameng Liu;Xinqi Sha;Changgao Huang;Lulu Hu;Shixiu Sun-
中科院分区：
文献类型：
--
作者：
Xin Tang;Shuang Zhao;Jieqiong Liu;Xiameng Liu;Xinqi Sha;Changgao Huang;Lulu Hu;Shixiu Sun-
背景技术背景：GSNOR（S-nitrosoglutathione reductase，S-亚硝基谷胱甘肽还原酶）作为一种S-亚硝基化的脱硝酶，在心肌重塑中具有保护心肌的作用，但GSNOR是否定位于其他细胞器并发挥新的作用尚不清楚。我们的目的是阐明线粒体GSNOR，一种新的亚细胞定位的GSNOR，对心脏重塑和心力衰竭（HF）的影响。方法：GSNOR的亚细胞定位采用细胞分级分离、免疫荧光染色和胶体金染色法。GSNOR在线粒体中的过表达是通过靶向RNA序列的腺相关病毒9实现的。GSNOR小鼠的心脏特异性敲除用于检查GSNOR在HF中的作用。利用生物素开关和液相色谱-串联质谱法对腺嘌呤核苷酸移位酶1（ANT 1）的S-亚硝基化位点进行了鉴定。结果：心衰患者心肌组织中GSNOR的表达受到抑制。一致的是，心脏特异性基因敲除小鼠表现出由横向主动脉缩窄引起的病理性重构加重。我们发现GSNOR也定位于线粒体。在血管紧张素II诱导的肥大心肌细胞中，线粒体GSNOR水平沿着线粒体功能受损而显著降低。心脏特异性基因敲除小鼠线粒体GSNOR水平的恢复显著改善了横主动脉缩窄诱导的HF小鼠的线粒体功能和心脏性能。从机制上讲，我们将ANT 1确定为GSNOR的直接目标。HF下线粒体GSNOR的降低导致半胱氨酸160（C160）处S-亚硝基化ANT 1的升高。根据这些发现，线粒体GSNOR或ANT 1 C160 A（非亚硝基化突变体）的过表达显著改善了线粒体功能，维持了线粒体膜电位，并上调了线粒体自噬。结论：我们发现了一种新的GSNOR定位于线粒体中，并发现线粒体GSNOR通过ANT 1去亚硝基化在维持线粒体稳态中起重要作用，这为HF提供了潜在的新治疗靶点。
BACKGROUND: The cardiac-protective role of GSNOR (S-nitrosoglutathione reductase) in the cytoplasm, as a denitrosylase enzyme of S-nitrosylation, has been reported in cardiac remodeling, but whether GSNOR is localized in other organelles and exerts novel effects remains unknown. We aimed to elucidate the effects of mitochondrial GSNOR, a novel subcellular localization of GSNOR, on cardiac remodeling and heart failure (HF). METHODS: GSNOR subcellular localization was observed by cellular fractionation assay, immunofluorescent staining, and colloidal gold particle staining. Overexpression of GSNOR in mitochondria was achieved by mitochondria-targeting sequence-directed adeno-associated virus 9. Cardiac-specific knockout of GSNOR mice was used to examine the role of GSNOR in HF. S-nitrosylation sites of ANT1 (adenine nucleotide translocase 1) were identified using biotin-switch and liquid chromatography-tandem mass spectrometry. RESULTS: GSNOR expression was suppressed in cardiac tissues of patients with HF. Consistently, cardiac-specific knockout mice showed aggravated pathological remodeling induced by transverse aortic constriction. We found that GSNOR is also localized in mitochondria. In the angiotensin II–induced hypertrophic cardiomyocytes, mitochondrial GSNOR levels significantly decreased along with mitochondrial functional impairment. Restoration of mitochondrial GSNOR levels in cardiac-specific knockout mice significantly improved mitochondrial function and cardiac performance in transverse aortic constriction–induced HF mice. Mechanistically, we identified ANT1 as a direct target of GSNOR. A decrease in mitochondrial GSNOR under HF leads to an elevation of S-nitrosylation ANT1 at cysteine 160 (C160). In accordance with these findings, overexpression of either mitochondrial GSNOR or ANT1 C160A, non-nitrosylated mutant, significantly improved mitochondrial function, maintained the mitochondrial membrane potential, and upregulated mitophagy. CONCLUSIONS: We identified a novel species of GSNOR localized in mitochondria and found mitochondrial GSNOR plays an essential role in maintaining mitochondrial homeostasis through ANT1 denitrosylation, which provides a potential novel therapeutic target for HF.