Study of the molecular basis of myocardial fibrosis is hampered by limited access to tissues from human patients and by confounding variables associated with sample accessibility, collection, processing and storage. Here, we report an integrative strategy based on mass spectrometry for the phosphoproteomic profiling of normal and fibrotic cardiac tissue obtained from surgical explants from patients with hypertrophic cardiomyopathy, from a transaortic-constriction mouse model of cardiac hypertrophy and fibrosis, and from a heart-on-a-chip model of cardiac fibrosis. We used the integrative approach to map the relative abundance of thousands of proteins, phosphoproteins and phosphorylation sites specific to each tissue source, to identify key signalling pathways driving fibrosis and to screen for anti-fibrotic compounds targeting glycogen synthase kinase 3, which has a consistent role as a key mediator of fibrosis in all three types of tissue specimen. The integrative disease-modelling strategy may reveal new insights into mechanisms of cardiac disease and serve as a test bed for drug screening.

心肌纤维化分子基础的研究因人类患者对组织的有限进入以及与样本可及性，收集，加工和存储相关的变量混杂而受阻。在这里，我们报告了一种基于质谱的整合策略，用于从肥大型心肌病患者的外科外植体，心脏肥大和纤维化的经主动脉收缩的小鼠模型以及心脏上提取的正常和纤维化心脏组织的磷酸化蛋白质组分析-心脏纤维化的芯片模型。我们使用整合方法来绘制每种组织来源特有的数千种蛋白质，磷蛋白和磷酸化位点的相对丰度，以确定驱动纤维化的关键信号通路，并筛选靶向糖原合酶激酶3的抗纤维化化合物，该化合物在所有三种类型的组织标本中均作为纤维化的关键介体具有一致的作用。综合疾病建模策略可能会揭示有关心脏病机制的新见解，并可以用作药物筛选的试验床。