A Systems Biology Approach To Cardiomyopathy in the D. Melanogaster Model System

描述（由申请人提供）：源于肌节和细胞结构突变的心肌病的特征是不良心脏重塑和易患心律失常和心源性猝死。病理生理学是复杂的，其基础的分子机制也是复杂的。系统地解决这种复杂性需要开发新的工具和模型。在这里，我们概述了一个多学科的系统生物学方法来研究心肌病的模型系统，利用快速遗传操作的力量，果蝇。作为心脏发育模型的长期价值，应用于心脏疾病的研究是其起步阶段。在一项技术突破性研究中，我们成功地利用蛋白质组学技术编制了果蝇心管的蛋白质纲要。使用生物信息学，我们表明，它在细胞成分，生物过程和分子功能的水平上具有心脏组织的标志。因此，我们已经开始评估限制性和扩张性心肌病所产生的肌节病变，使用定量蛋白质组学方法在果蝇模型的心脏重塑。 该建议概述了我们的系统生物学方法，通过该方法，蛋白质表达和翻译后修饰的配置文件被翻译为可检验的假设，通过应用生物信息学本体论和网络分析。蛋白质网络的扰动将与哺乳动物综合兴奋-收缩耦合和生物能量学（ECME）数学模型的模拟交叉引用。网络/模型模拟之后将进行异常功能的生理学评估（例如生物能量学、Ca 2+处理）。初步分析表明，限制性心肌病突变Mhc 5表现出实质性的代谢重塑，这表明腺苷一磷酸活化蛋白激酶（AMPK）的可能作用。我们概述了我们正在追求的新技术创新，作为衡量果蝇心脏性能的武器库的一部分。这项探索性/开发资助的直接2年目标是确定一组新的候选蛋白或PTM特征，这些蛋白或PTM特征导致限制性和扩张性心肌病果蝇模型的生物活性改变。简而言之，我们概述了一种策略，以简化搜索限制性和扩张型心肌病的遗传抑制因子，即未来快速靶向敲除和过表达的靶点。这些新的修饰果蝇蛋白的人类同源物可能代表治疗药物设计的新靶点。

DESCRIPTION (provided by applicant): Cardiomyopathies that stem from sarcomeric and cytostructural mutations are characterized by adverse cardiac remodeling and predisposition to both arrhythmia and sudden cardiac death. The pathophysiology is complex, as are molecular mechanisms that underlie it. Tackling this complexity systematically demands the development of new tools and models. Here, we outline a multidisciplinary systems biology approach to the study of cardiomyopathy in a model system that leverages the power of rapid genetic manipulation, Drosophila melanogaster. Long valued as a model of cardiac development, application to the study of cardiac disease is its infancy. In a technical breakthrough study, we have succeeded in using proteomic techniques to compile a protein compendium of the Drosophila cardiac tube. Using bioinformatics, we show that it bears hallmarks of heart tissue at the level of cellular componentry, biological processes and molecular functions. We have therefore begun to assess cardiac remodeling in Drosophila models of restrictive and dilated cardiomyopathy arising from sarcomeric lesions using quantitative proteomic methods. This proposal outlines our systems biology approach, by which profiles of protein expression and post-translational modification are translated to testable hypotheses by the application of bioinformatic ontological and network analyses. The perturbation of protein networks will be cross-referenced with simulations from mathematical models of mammalian integrated excitation-contraction coupling and bioenergetics (ECME). Network/Model simulations will be followed up with physiological assessments of aberrant function (e.g. bioenergetics, Ca2+ handling). Preliminary analysis indicates the restrictive cardiomyopathy mutant Mhc5 shows substantial metabolic remodeling, suggesting a possible role for adenosine monophosphate activated protein kinase (AMPK). We outline new technical innovations that we are pursuing as part of the arsenal to measure Drosophila heart performance. The immediate 2-year goal of this exploratory/development grant is to identify a set of novel protein candidates or PTM signatures that lead to altered biological activity in Drosophila models of restrictive and dilated cardiomyopathy. In short, we outline a strategy to streamline the search for genetic suppressors of restrictive and dilated cardiomyopathy i.e. targets for future rapid targeted knockdown and overexpression. Human homologues of these novel modified drosophila proteins may represent novel targets for therapeutic drug design.