Key Laboratory of Cardiovascular and Cerebrovascular Medicine; Key Laboratory of Targeted Intervention of Cardiovascular Disease; Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Jiangsu, China (Y.Z., H.Z., S.Z., Z.Q., Y.H., X.Z., W.W., K.Y., L.H., S.S., F.C., L.X., Y.J.).
Department of Thoracic and Cardiovascular Surgery, Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing Drum Tower Hospital Clinical College of Nanjing Medical University, Institute of Cardiothoracic Vascular Disease, Nanjing University, China (X.T., Q.Z., D.W.).
Circulation. 2024 Jun 11;149(24):1903-1920. doi: 10.1161/CIRCULATIONAHA.123.066404. Epub 2024 Feb 15.
S-Nitrosylation (SNO), a prototypic redox-based posttranslational modification, is involved in cardiovascular disease. Aortic aneurysm and dissection are high-risk cardiovascular diseases without an effective cure. The aim of this study was to determine the role of SNO of Septin2 in macrophages in aortic aneurysm and dissection.
Biotin-switch assay combined with liquid chromatography-tandem mass spectrometry was performed to identify the S-nitrosylated proteins in aortic tissue from both patients undergoing surgery for aortic dissection and mice infused with angiotensin II. Angiotensin II-induced aortic aneurysm model and β-aminopropionitrile-induced aortic aneurysm and dissection model were used to determine the role of SNO of Septin2 (SNO-Septin2) in aortic aneurysm and dissection development. RNA-sequencing analysis was performed to recapitulate possible changes in the transcriptome profile of SNO-Septin2 in macrophages in aortic aneurysm and dissection. Liquid chromatography-tandem mass spectrometry and coimmunoprecipitation were used to uncover the TIAM1-RAC1 (Ras-related C3 botulinum toxin substrate 1) axis as the downstream target of SNO-Septin2. Both R-Ketorolac and NSC23766 treatments were used to inhibit the TIAM1-RAC1 axis.
Septin2 was identified S-nitrosylated at cysteine 111 (Cys111) in both aortic tissue from patients undergoing surgery for aortic dissection and mice infused with Angiotensin II. SNO-Septin2 was demonstrated driving the development of aortic aneurysm and dissection. By RNA-sequencing, SNO-Septin2 in macrophages was demonstrated to exacerbate vascular inflammation and extracellular matrix degradation in aortic aneurysm. Next, TIAM1 (T lymphoma invasion and metastasis-inducing protein 1) was identified as a SNO-Septin2 target protein. Mechanistically, compared with unmodified Septin2, SNO-Septin2 reduced its interaction with TIAM1 and activated the TIAM1-RAC1 axis and consequent nuclear factor-κB signaling pathway, resulting in stronger inflammation and extracellular matrix degradation mediated by macrophages. Consistently, both R-Ketorolac and NSC23766 treatments protected against aortic aneurysm and dissection by inhibiting the TIAM1-RAC1 axis.
SNO-Septin2 drives aortic aneurysm and dissection through coupling the TIAM1-RAC1 axis in macrophages and activating the nuclear factor-κB signaling pathway-dependent inflammation and extracellular matrix degradation. Pharmacological blockade of RAC1 by R-Ketorolac or NSC23766 may therefore represent a potential treatment against aortic aneurysm and dissection.
S-亚硝基化（SNO）是一种典型的基于氧化还原的翻译后修饰，参与心血管疾病。胸主动脉瘤和夹层是一种高风险的心血管疾病，目前尚无有效的治疗方法。本研究旨在确定 SNO 修饰的 Septin2 在巨噬细胞中的作用，进而阐明其在胸主动脉瘤和夹层中的作用。
采用生物素开关分析联合液相色谱-串联质谱技术，鉴定接受胸主动脉夹层手术的患者和接受血管紧张素 II 输注的小鼠主动脉组织中的 S-亚硝基化蛋白。采用血管紧张素 II 诱导的胸主动脉瘤模型和β-氨基丙腈诱导的胸主动脉瘤和夹层模型，确定 SNO-Septin2（SNO- Septin2）在胸主动脉瘤和夹层发展中的作用。采用 RNA 测序分析，重现 SNO-Septin2 在巨噬细胞中参与胸主动脉瘤和夹层的转录组谱的可能变化。采用液相色谱-串联质谱和免疫共沉淀技术，揭示 TIAM1-RAC1（Ras 相关 C3 肉毒梭菌毒素底物 1）轴作为 SNO-Septin2 的下游靶标。采用 R-酮咯酸和 NSC23766 治疗来抑制 TIAM1-RAC1 轴。
在接受胸主动脉夹层手术的患者的主动脉组织和接受血管紧张素 II 输注的小鼠的主动脉组织中，均鉴定到 Cys111 位点的 Septin2 发生 SNO 修饰。SNO-Septin2 促进了胸主动脉瘤和夹层的发展。通过 RNA 测序分析，发现巨噬细胞中的 SNO-Septin2 可加剧主动脉瘤中的血管炎症和细胞外基质降解。进一步鉴定到 T 淋巴瘤入侵和转移诱导蛋白 1（TIAM1）是 SNO-Septin2 的靶蛋白。在机制上，与未修饰的 Septin2 相比，SNO-Septin2 降低了与 TIAM1 的相互作用，激活了 TIAM1-RAC1 轴和随后的核因子-κB 信号通路，导致巨噬细胞介导的炎症和细胞外基质降解增强。同样，R-酮咯酸和 NSC23766 治疗通过抑制 TIAM1-RAC1 轴，可预防胸主动脉瘤和夹层。
SNO-Septin2 通过在巨噬细胞中偶联 TIAM1-RAC1 轴并激活核因子-κB 信号通路依赖性炎症和细胞外基质降解，驱动胸主动脉瘤和夹层的发生。R-酮咯酸或 NSC23766 抑制 RAC1 可能代表治疗胸主动脉瘤和夹层的一种潜在方法。
