CHANGES IN THE RBC PROTEOME DURING HEALTH AND DISEASE

摘要：最近对红细胞（RBC）蛋白质组的研究已经产生了广泛的膜蛋白和可溶性蛋白列表。虽然这些红细胞蛋白的目录相当令人印象深刻，但它们几乎没有包含任何定量信息。差异蛋白表达的分析对于发现和理解疾病的生物学基础，以及为诊断或预后提供新的生物标志物，评估风险特征和结果，以及确定个性化治疗的新靶点至关重要。在过去的几年中，质谱法和用于复杂多肽混合物定量比较分析的软件取得了重大进展。我们假设红细胞中蛋白质的全球定量表达模式将反映正常生活中存在的差异，但也会反映因疾病而发生的差异。在这里，我们建议开发一个强大的，无标记的蛋白质组学平台来定义红细胞中的差异蛋白表达。我们将评估技术变异的水平，并定义由于个体间和性别差异而产生的变异。然后，我们将测试开发的平台，并将其应用于定义两种经典红细胞疾病，阵发性夜间血红蛋白尿（PNH）和Diamond Blackfan贫血（DBA）中全局蛋白表达模式的差异。最后，我们将使用开发的平台和定义疾病的特征蛋白来监测在疾病特异性治疗期间红细胞蛋白质组发生的变化。我们的发现可能会为这些疾病的病理生理学提供新的见解，并可能导致新的标记物的鉴定，或发现诊断测试或药物开发的新靶点。此外，我们开发的工具应该被证明广泛适用于不同生理或疾病状态的红细胞膜蛋白质组的定量比较。

DESCRIPTION (provided by applicant): "Changes in the RBC proteome during health and disease" Abstract: Recent studies of the red blood cell (RBC) proteome have yielded extensive lists of both membrane and soluble proteins. While these catalogs of RBC proteins are quite impressive, they contain little if any quantitative information. The analysis of differential protein expression can be crucial for the discovery and understanding of the biological underpinnings of disease, as well as for providing novel biomarkers for diagnosis or prognosis, for assessing risk profiles and outcomes, and for identifying novel targets for individualized treatment. Within the last few years there have been significant advances in mass spectrometry and software for the quantitative, comparative analysis of complex mixtures of peptides. We hypothesize that the global quantitative expression patterns of proteins in RBC will reflect differences that exist in normal live but also those that occur due to disease. Here we propose to develop a robust, label-free proteomics platform for defining differential protein expression in RBCs. We will assess the level of technical variation, as well as define variations due to inter-individual and gender differences. We will then test the platform developed and apply it to define the differences in global protein expression patterns in two classic red blood cell disorders, Paroxysmal Nocturnal Hemoglobinuria (PNH) and Diamond Blackfan Anemia (DBA). Finally, we will use the platform developed and the signature proteins that define the disease to monitor the changes that occur in the RBC proteome during disease specific treatment. Our findings are likely to provide new insights into the pathophysiology of these diseases, and might lead to the identification of novel markers, or to the discovery of new targets for the development of diagnostic tests or drugs. Furthermore, the tools we develop should prove broadly applicable to the quantitative comparison of RBC membrane proteomes of varying physiological or disease states. PUBLIC HEALTH RELEVANCE: We hypothesize that the global quantitative expression patterns of red cell proteins will reflect normal physiological variations, as well as variations that occur during disease. In this grant we propose to develop a robust, label-free proteomics platform for defining differential protein expression in red blood cells. We will assess the level of technical variation associated with the platform and define variations due to normal physiological differences. We will then test the developed platform by applying it to define the differences in global protein expression patterns that occur in two classic red blood cell disorders, namely Paroxysmal Nocturnal Hemoglobinuria (PNH) and Diamond Blackfan Anemia (DBA). Our investigations are likely to provide new insights into the pathophysiology of these diseases, and might lead to the identification of novel markers, or to the discovery of new targets for the development of diagnostic tests or drugs. Furthermore, the platform we develop should prove broadly applicable to the quantitative comparison of RBC proteomes of varying physiological or disease states.