S-Nitrosylation of Septin2 Exacerbates Aortic Aneurysm and Dissection by Coupling the TIAM1-RAC1 Axis in Macrophages

Yan Zhang; Hao Zhang; Shuang Zhao; Zhenhua Qi; Yiwei He; Xuhong Zhang; Wencheng Wu; Ke Yan; Lulu Hu; Shixiu Sun; Xinlong Tang; Qing Zhou; Feng Chen; Aihua Gu; Liansheng Wang; Zhiren Zhang; Bo Yu; Dongjin Wang; Yi Han; Liping Xie; Yong Ji

doi:10.1161/CIRCULATIONAHA.123.066404

S-Nitrosylation of Septin2 Exacerbates Aortic Aneurysm and Dissection by Coupling the TIAM1-RAC1 Axis in Macrophages

Circulation. 2024 Feb 15. doi: 10.1161/CIRCULATIONAHA.123.066404. Online ahead of print.

Authors

Yan Zhang^#¹, Hao Zhang^#^{1

2}, Shuang Zhao^#¹, Zhenhua Qi^{1

2}, Yiwei He^{1

2}, Xuhong Zhang^{1

2}, Wencheng Wu^{1

2}, Ke Yan^{1

2}, Lulu Hu^{1

2}, Shixiu Sun^{1

2}, Xinlong Tang³, Qing Zhou³, Feng Chen^{1

2

4}, Aihua Gu⁵, Liansheng Wang⁶, Zhiren Zhang⁷, Bo Yu⁸, Dongjin Wang³, Yi Han⁹, Liping Xie^{1

2}, Yong Ji^{1

2

10

7}

Affiliations

¹ 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.).
² Medical Basic Research Innovation Center for Cardiovascular and Cerebrovascular Diseases, Ministry of Education, Beijing, China (H.Z., Z.Q., Y.H., X.Z., W.W., K.Y., L.H., S.S., F.C., L.X., Y.J.).
³ 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.).
⁴ Department of Forensic Medicine, Nanjing Medical University, China. (F.C.).
⁵ School of Public Health, Nanjing Medical University, China. (A.G.).
⁶ Departments of Cardiology, First Affiliated Hospital of Nanjing Medical University, China (L.W.).
⁷ State Key Laboratory of Frigid Zone Cardiovascular Diseases (SKLFZCD), Harbin Medical University, Heilongjiang, PR China (Z.Z., Y.J.).
⁸ Department of Cardiology, Second Affiliated Hospital of Harbin Medical University, Key Laboratory of Myocardial Ischemia, Ministry of Education, Harbin Medical University, Heilongjiang, China (B.Y.).
⁹ Department of Geriatrics, the First Affiliated Hospital of Nanjing Medical University, China (Y.H.).
¹⁰ Gusu School, Nanjing Medical University, Suzhou, China (Y.J.).

^# Contributed equally.

PMID: 38357802
DOI: 10.1161/CIRCULATIONAHA.123.066404

Abstract

Background: 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.

Methods: 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 Apoe^-/- 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.

Results: Septin2 was identified S-nitrosylated at cysteine 111 (Cys111) in both aortic tissue from patients undergoing surgery for aortic dissection and Apoe^-/- 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.

Conclusions: 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.

Keywords: aortic aneurysm; aortic dissection; macrophages.