School of Pharmaceutical Sciences, National and Local United Engineering Lab of Druggability and New Drugs Evaluation, Sun Yat-sen University, Guangzhou, P. R. China.
Aab Cardiovascular Research Institute, University of Rochester School of Medicine and Dentistry, Rochester, NY, USA.
Cell Death Differ. 2021 Oct;28(10):2871-2887. doi: 10.1038/s41418-021-00789-w. Epub 2021 May 4.
Sorting nexins (SNXs), the retromer-associated cargo binding proteins, have emerged as critical regulators of the trafficking of proteins involved in the pathogenesis of diverse diseases. However, studies of SNXs in the development of cardiovascular diseases, especially cardiac hypertrophy and heart failure, are lacking. Here, we ask whether SNX3, the simplest structured isoform in the SNXs family, may act as a key inducer of myocardial injury. An increased level of SNX3 was observed in failing hearts from human patients and mice. Cardiac-specific Snx3 knockout (Snx3-cKO) mice and Snx3 transgenic (Snx3-cTg) mice were generated to evaluate the role of Snx3 in myocardial hypertrophy, fibrosis, and heart function by morphology, echocardiography, histological staining, and hypertrophic biomarkers. We report that Snx3-cKO in mice significantly protected against isoproterenol (ISO)-induced cardiac hypertrophy at 12 weeks. Conversely, Snx3-cTg mice were more susceptible to ISO-induced cardiac hypertrophy at 12 weeks and showed aggravated cardiac injury even heart failure at 24 weeks. Immunoprecipitation-based mass spectrometry, immunofluorescent staining, co-immunoprecipitation, localized surface plasmon resonance, and proximity ligation assay were performed to examine the direct interaction of SNX3-retromer with signal transducer and activator of transcription 3 (STAT3). We discovered that STAT3 was a new interacting partner of SNX3-retromer, and SNX3-retromer served as an essential platform for assembling gp130/JAK2/STAT3 complexes and subsequent phosphorylation of STAT3 by direct combination at EE. SNX3-retromer and STAT3 complexes were transiently imported into the nucleus after hypertrophic stimuli. The pharmacological inhibition or knockdown of STAT3 reversed SNX3 overexpression-induced myocardial injury. STAT3 overexpression blunts the beneficial function of SNX3 knockdown on hypertrophic cardiomyocytes. We show that SNX3-retromer promoted importin α3-mediated STAT3 nuclear trafficking and ultimately leading to cardiac injury. Taken together, our study reveals that SNX3 plays a key role in cardiac function and implicates SNX3 as a potential therapeutic target for cardiac hypertrophy and heart failure.
分选连接蛋白(SNXs)是与逆转运体相关的货物结合蛋白,已成为参与多种疾病发病机制的蛋白质运输的关键调节因子。然而,SNXs 在心血管疾病,特别是心肌肥大和心力衰竭发展中的研究还很缺乏。在这里,我们研究了 SNXs 家族中最简单结构的亚型 SNX3 是否可能作为心肌损伤的关键诱导因子。在人类患者和小鼠的衰竭心脏中观察到 SNX3 水平升高。通过形态学、超声心动图、组织学染色和肥大标志物,生成了心脏特异性 Snx3 敲除(Snx3-cKO)小鼠和 Snx3 转基因(Snx3-cTg)小鼠,以评估 Snx3 在心肌肥大、纤维化和心脏功能中的作用。我们报告说,在 12 周时,Snx3-cKO 显著保护小鼠免受异丙肾上腺素(ISO)诱导的心肌肥大。相反,在 12 周时,Snx3-cTg 小鼠对 ISO 诱导的心肌肥大更敏感,甚至在 24 周时出现更严重的心脏损伤和心力衰竭。通过免疫沉淀质谱分析、免疫荧光染色、共免疫沉淀、局部表面等离子体共振和邻近连接分析,研究了 SNX3-逆转运体与信号转导和转录激活因子 3(STAT3)的直接相互作用。我们发现 STAT3 是 SNX3-逆转运体的一个新的相互作用伙伴,SNX3-逆转运体作为组装 gp130/JAK2/STAT3 复合物的必需平台,并通过 EE 处的直接结合对 STAT3 进行后续磷酸化。在肥大刺激后,SNX3-逆转运体和 STAT3 复合物会被瞬时导入细胞核。STAT3 的药理学抑制或敲低可逆转 SNX3 过表达诱导的心肌损伤。STAT3 过表达削弱了 SNX3 敲低对肥大心肌细胞的有益作用。我们表明,SNX3-逆转运体促进了进口蛋白 α3 介导的 STAT3 核转运,最终导致了心脏损伤。总之,我们的研究揭示了 SNX3 在心脏功能中的关键作用,并暗示 SNX3 可能是心肌肥大和心力衰竭的潜在治疗靶点。