冠心病代谢组学特征谱和生物学调控机制研究

Metabolic abnormalities play an important role in the presence and development of coronary artery disease (CAD). Pathogenesis and diagnostic biomarkers for diseases can be discovered by metabolomic profiling of human fluids. If the various types of CAD can be accurately characterized by metabolomics, effective treatment may be targeted without using unnecessary therapies and resources. In our previous work, we collected large-scale and multicenter (n=4) plasma samples from 2324 CAD patients who underwent invasive coronary angiography. For the first time, we reported the comprehensive metabolomic characterization of CAD (Journal of the American College of Cardiology, 2016, 68(12):1281-1293. Results showed that plasma metabolomics are powerful in characterizing metabolic disturbances. Differences in small-molecule metabolites may reflect underlying CAD and serve as biomarkers for CAD progression. As a series of work, this project aims to study metabolomic profile to assess the diagnostic and prognostic values of metabolomics-based biomarkers in different types of CAD. The major contents include: (1) A large-cohort of retrospective and prospective CAD plasma samples (n>20000) will be collected in parallel with completed baseline information; (2) Novel metabolomic strategies will be developed towards high sensitivity, high resolution and visualization, incorporating liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, and electrochemiluminescence imaging methods; (3) The metabolomic profiling for Chinese CAD patients will be characterized. The diagnostic and prognostic values of metabolomics-based biomarkers will be assessed, and will be validated in large-scale and multicenter samples. (4) The mechanisms by which metabolites (2-3 significant metabolites) elevate or decrease will be investigated using multidisciplinary methods of biochemistry, cell biology and molecular biology. The functions of significantly differential metabolites associated with the presence and severity of CAD will be also investigated. The achievements from this project will be of significance in early diagnosis, disease differentiation, prognostic evaluation, and drugs’ response.

代谢异常在冠心病发生发展中扮演重要角色，代谢组学可揭示冠心病代谢紊乱网络和关键代谢标志物，为其诊断和治疗提供潜在靶点。前期研究中，基于临床血浆标本（n=2324)，首次绘制了冠心病及其不同临床分型的代谢组学特征谱，发现若干新代谢标志物(J Am Coll Cardiol, 2016)。本项目拟在已有研究基础上，构建临床资料完备的冠心病回顾性和前瞻性人群队列(n>20000)；融合液质、气质联用和电化学发光成像技术，创建超灵敏、高覆盖、可视化的代谢组定性定量分析技术体系；建立中国人群冠心病的代谢组学特征谱，发现、筛选和鉴定一批用于冠心病早期诊断和预后评价的代谢标志物，并开展大规模多中心临床验证；综合运用生物化学、细胞/分子生物学等方法深入研究2-3个代谢标志物失调的机制以及在冠心病发生发展中的功能与生物学意义。研究成果将为冠心病的早期诊断、精确分型、预后评价和药物反应性等提供重要理论指导。

依托“冠心病代谢组学特征谱和生物学调控机制研究”项目（No.91639115），成立全国首个临床代谢组学中心（中国药科大学），旨在以临床大样品为基石、以代谢组学为手段，揭示疾病潜在的诊疗生物标志物和可干预靶标；与江苏武进医院、北京阜外医院、江苏省人民医院等合作，建立了心血管临床多中心大样本库(n=30000)；建立了针对脂质、胆汁酸、氨基酸、糖类等内源性代谢物的靶向、非靶向代谢组学研究系列方法；绘制了冠心病及其不同临床分型的血浆代谢组学特征谱，鉴定了与冠心病发生发展相关的差异代谢标志物125个，主要包括磷脂相关代谢通路抑制、氨基酸代谢增强、酰基肉碱类代谢通路激活、胆汁酸代谢减弱等多种代谢紊乱特征；进一步从表型代谢组学深入到功能代谢组学，对代谢标志物N-乙酰神经氨酸（Neu5Ac）、琥珀酸、游离脂肪酸、腺苷-3’,5’-环化-磷酸（cAMP）等在冠心病中的功能和机制开展深入研究，例如，发现Neu5Ac堆积可选择性结合Rho家族中的 RhoA、Cdc42 而非Rac蛋白，进而激活Rho-ROCK信号通路诱发心肌损伤；以代谢标志物的上下游关键调控酶为干预靶标，采用基因编辑、药理抑制剂，证实流感病毒神经氨酸酶抑制剂药物（奥司他韦和扎那米韦）可通过抑制代谢物Neu5Ac堆积减少心肌缺血损伤，发现人参皂苷可通过调节p52-PDE4B互作减少代谢标志物cAMP的水平进而对抗糖脂代谢紊乱作用。项目共发表SCI论文7篇，包括Circulation、Nat Commun等，研究成果写入“爆发性心肌炎诊治英文版专家共识”，在长城心脏病学会议、钱江国际心血管病会议作邀请报告4次；获美国授权专利1项、中国发明专利6项，部分专利已转化，为市民提供冠心病早期预警，具有无创、便捷优势；培养了一支多学科交叉的心血管基础-临床结合研究队伍，项目负责人获得国家杰出青年科学基金、中国青年科技奖，培养博士4名。