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

描述（由申请人提供）：由肌瘤和细胞结构突变引起的心肌病的特征是不良的心脏重构和心律失常和心源性猝死的易感性。病理生理学是复杂的，其分子机制也是复杂的。系统地处理这种复杂性需要开发新的工具和模型。在这里，我们概述了一种多学科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，黑腹果蝇。长期以来，它一直被视为心脏发育的一种模式，但在心脏病研究中的应用还处于起步阶段。在一项突破性的技术研究中，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。利用生物信息学，我们表明它在细胞成分、生物过程和分子功能水平上具有心脏组织的特征。因此，我们开始使用定量蛋白质组学方法评估由肌瘤病变引起的限制性和扩张性心肌病果蝇模型中的心脏重塑。本提案概述了我们的系统生物学方法，通过应用生物信息学本体论和网络分析，将蛋白质表达和翻译后修饰的概况转化为可测试的假设。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）的数学模型的模拟交叉参考。网络/模型模拟将随后进行异常功能的生理评估（例如生物能量学，Ca2+处理）。初步分析表明，限制性心肌病突变体Mhc5表现出实质性的代谢重塑，这可能与腺苷单磷酸活化蛋白激酶（AMPK）有关。我们概述了我们正在追求的新技术创新，作为衡量果蝇心脏表现的武器库的一部分。这项探索性/开发性资助的近期2年目标是确定一组新的候选蛋白或PTM特征，这些候选蛋白或PTM特征导致限制性和扩张型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，以简化限制性和扩张型心肌病基因抑制因子的搜索，即未来快速靶向敲除和过表达的目标。这些新修饰的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.