Lei Zhang1, Ting-Ting Wei1, Yong Li2, Jing Li3, Yong Fan1, Feng-Qing Huang1, Yuan-Yuan Cai3, Gaoxiang Ma3, Jin-Feng Liu1, Qian-Qian Chen1, Shi-Lei Wang1, Honglin Li4, Raphael N Alolga1, Baolin Liu3, Dong-Sheng Zhao5, Jian-Hua Shen6, Xiang-Ming Wang7, Wei Zhu8, Ping Li9, Lian-Wen Qi9,3. 1. State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing (L.Z., T.-T.W., Y.F., F.-Q.H., J.-F.L., Q.-Q.C., S.-L.W., R.N.A., P.L., L.-W.Q.). 2. Department of Cardiology, the Affiliated Wujin Hospital of Jiangsu University, Changzhou, China (Y.L.). 3. Clinical Metabolomics Center, China Pharmaceutical University, Nanjing (J.L., Y.-Y.C., G.M., B.L., L.-W.Q.). 4. Shanghai Key Laboratory of New Drug Design, State Key Laboratory of Bioreactor Engineering, School of Pharmacy, East China University of Science and Technology (H.L.). 5. Department of Cardiology, Second Affiliated Hospital of Nantong University, Nantong (D.-S.Z.). 6. Department of Cardiology, Northern Jiangsu People's Hospital, Yangzhou, China (J.-H.S.). 7. Department of Geriatric Cardiology (X.-M.W.). 8. Department of Oncology (W.Z.), First Affiliated Hospital of Nanjing Medical University, China liping2004@126.com Qilw@cpu.edu.cn zhuwei@njmu.edu.cn. 9. State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing (L.Z., T.-T.W., Y.F., F.-Q.H., J.-F.L., Q.-Q.C., S.-L.W., R.N.A., P.L., L.-W.Q.) liping2004@126.com Qilw@cpu.edu.cn zhuwei@njmu.edu.cn.
Abstract
BACKGROUND: As new biomarkers of coronary artery diseases (CAD) emerge via metabolomics, the underlying functional mechanisms remain to be elucidated. Functional metabolomics aims to translate metabolomics-derived biomarkers to disease mechanisms. METHODS: A cohort of 2324 patients who underwent coronary angiography from 4 independent centers was studied. A combination of ultra-performance liquid chromatography and quadrupole time-of-flight mass spectrometry in the negative ion mode was used for untargeted analysis of metabolites in plasma. Significant differential metabolites were identified by cross-comparisons with and within CAD types, including normal coronary artery, nonobstructvie coronary atherosclerosis, stable angina, unstable angina, and acute myocardial infarction. A tandem liquid chromatography-mass spectrometry-based approach using isotope-labeled standard addition was subsequently performed for targeted analysis of the metabolic marker N-acetylneuraminic acid (Neu5Ac). A functional metabolomics strategy was proposed to investigate the role of Neu5Ac in the progression of CAD by using in vitro and in vivo models. RESULTS: We identified a total of 36 differential metabolites, 35 of which were confirmed with reference compounds. Elevation of Neu5Ac was observed in plasma during CAD progression in center 1 (P=4.0e-64, n=2019) and replicated in 3 independent centers (n=305). The increased level of Neu5Ac in plasma was confirmed by accurate targeted quantification. Mechanistically, Neu5Ac was able to trigger myocardial injury in vitro and in vivo by activation of the Rho/Rho-associated coiled-coil containing protein kinase signaling pathway through binding to RhoA and Cdc42, but not Rac1. Silencing neuraminidase-1, the enzyme that regulates Neu5Ac generation, ameliorated oxygen-glucose deprivation-induced injury in cardiomyocytes and ligation/isoprenaline-induced myocardial ischemia injury in rats. Pharmacological inhibition of neuraminidase by anti-influenza drugs, oseltamivir and zanamivir, also protected cardiomyocytes and the heart from myocardial injury. CONCLUSIONS: Functional metabolomics identified a key role for Neu5Ac in acute myocardial infarction, and targeting neuraminidase-1 may represent an unrecognized therapeutic intervention for CAD.
BACKGROUND: As new biomarkers of coronary artery diseases (CAD) emerge via metabolomics, the underlying functional mechanisms remain to be elucidated. Functional metabolomics aims to translate metabolomics-derived biomarkers to disease mechanisms. METHODS: A cohort of 2324 patients who underwent coronary angiography from 4 independent centers was studied. A combination of ultra-performance liquid chromatography and quadrupole time-of-flight mass spectrometry in the negative ion mode was used for untargeted analysis of metabolites in plasma. Significant differential metabolites were identified by cross-comparisons with and within CAD types, including normal coronary artery, nonobstructvie coronary atherosclerosis, stable angina, unstable angina, and acute myocardial infarction. A tandem liquid chromatography-mass spectrometry-based approach using isotope-labeled standard addition was subsequently performed for targeted analysis of the metabolic marker N-acetylneuraminic acid (Neu5Ac). A functional metabolomics strategy was proposed to investigate the role of Neu5Ac in the progression of CAD by using in vitro and in vivo models. RESULTS: We identified a total of 36 differential metabolites, 35 of which were confirmed with reference compounds. Elevation of Neu5Ac was observed in plasma during CAD progression in center 1 (P=4.0e-64, n=2019) and replicated in 3 independent centers (n=305). The increased level of Neu5Ac in plasma was confirmed by accurate targeted quantification. Mechanistically, Neu5Ac was able to trigger myocardial injury in vitro and in vivo by activation of the Rho/Rho-associated coiled-coil containing protein kinase signaling pathway through binding to RhoA and Cdc42, but not Rac1. Silencing neuraminidase-1, the enzyme that regulates Neu5Ac generation, ameliorated oxygen-glucosedeprivation-induced injury in cardiomyocytes and ligation/isoprenaline-induced myocardial ischemia injury in rats. Pharmacological inhibition of neuraminidase by anti-influenza drugs, oseltamivir and zanamivir, also protected cardiomyocytes and the heart from myocardial injury. CONCLUSIONS: Functional metabolomics identified a key role for Neu5Ac in acute myocardial infarction, and targeting neuraminidase-1 may represent an unrecognized therapeutic intervention for CAD.
Authors: Ina Nemet; Prasenjit Prasad Saha; Nilaksh Gupta; Weifei Zhu; Kymberleigh A Romano; Sarah M Skye; Tomas Cajka; Maradumane L Mohan; Lin Li; Yuping Wu; Masanori Funabashi; Amanda E Ramer-Tait; Sathyamangla Venkata Naga Prasad; Oliver Fiehn; Federico E Rey; W H Wilson Tang; Michael A Fischbach; Joseph A DiDonato; Stanley L Hazen Journal: Cell Date: 2020-03-05 Impact factor: 41.582