Soluble protein markers of T1D progression.

T1D 进展的可溶性蛋白标记物。

• 批准号: 6951208
• 负责人: Paul E Harris
• 金额: $ 25.61万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6951208
• 关键词: autoantibody; biomarker; blood proteins; clinical research; human tissue; immunologic assay /test; insulin dependent diabetes mellitus; laboratory rat; mass spectrometry; myocardial ischemia /hypoxia; myocardium; pancreatic islets; pathologic process; posttranslational modifications; serum; technology /technique development

DESCRIPTION (provided by applicant): Prediction methods for the progression of type 1 diabetes (T1D), based on current genetic and autoantibody testing, lack sensitivity and specificity. In this R21 proposal we describe proteomics based methods of study aimed at the identification and detection of specific markers of beta cell destruction and active anti islet immunity. The overall strategy of this proposal is to continue our efforts in mapping tissue specific, tissue restricted, and disease induced gene products of human islets. From previous work we have at our disposal a map of human islet tissue restricted transcripts. In collaboration with experts in mass spectroscopy of proteins (Dr. Caputos' group), we now propose to take advantage of this unique information to enhance studies of soluble human islet proteins with specific applications in mind. We propose that the study of the proteome of purified human islets and those cultured in interferon alpha ex vivo, will lead to the identification of tissue restricted and/or disease associated proteins, some of which will represent potentially useful serum markers of nascent type 1 diabetes and/or its progression. We hypothesize that some of these markers will represent by-products of inflammatory processes that precede and/or drive beta cell destruction, where as other markers, representing proteins either specific to or highly restricted to islet tissue, will provide evidence of beta cell damage or correlate with beta cell mass. We propose that due to beta cell destruction, some of these markers may be present in serum in forms that reflect incomplete post-translational modification. Drawing from the model of release of cardiac muscle specific proteins following myocardial ischemia, in Specific Aim One, we propose to study serum proteins in the rat BB model of human T1D with the goal of determining whether islet specific proteins are released into circulation following the destruction of beta cells. Using informatics tools and traditional protein sequencing methods we will identify serum proteins and peptides that correlate with beta cell destruction and diabetes. In Specific aim two, we will study the proteome of purified human islets, with the goals of identifying the most prevalent islet tissue specific proteins, with a particular focus on identifying their levels of post translational modification. In parallel, we will develop a SELDI-TOF based immunoassay chip, composed of antibodies recognizing beta cell specific, neuroendocrine tissue specific and/or islet inflammation-associated protein markers. The choice of markers is based on our knowledge of abundant islet transcripts and information we obtain from our islet proteome characterization. Having established the performance characteristics of the Islet chip, we will return to the rat model and retrospectively search for these markers in serum and determine their association with beta cell damage and diabetes. We have drawn together an interdisciplinary research team with the common goal of developing of markers of the destruction of beta cells of the endocrine pancreas.

描述（由申请人提供）： 基于当前基因和自身抗体检测的 1 型糖尿病 (T1D) 进展预测方法缺乏敏感性和特异性。在这个 R21 提案中，我们描述了基于蛋白质组学的研究方法，旨在识别和检测 β 细胞破坏和主动抗胰岛免疫的特定标记物。该提案的总体策略是继续努力绘制人类胰岛的组织特异性、组织限制性和疾病诱导的基因产物。根据之前的工作，我们可以使用人类胰岛组织限制转录本的图谱。通过与蛋白质质谱专家（Caputos 博士的小组）合作，我们现在建议利用这一独特的信息来加强对可溶性人类胰岛蛋白的研究，并考虑到特定的应用。我们建议，对纯化的人胰岛以及在干扰素α离体中培养的胰岛的蛋白质组进行研究，将导致组织限制性和/或疾病相关蛋白的鉴定，其中一些将代表新生1型糖尿病的潜在有用的血清标志物和/或其进展。我们假设其中一些标记物将代表先于和/或驱动β细胞破坏的炎症过程的副产品，而代表胰岛组织特异性或高度限制性蛋白质的其他标记物将提供β细胞损伤或与 β 细胞质量相关。我们认为，由于β细胞的破坏，其中一些标记物可能以反映不完整的翻译后修饰的形式存在于血清中。借鉴心肌缺血后心肌特异性蛋白释放的模型，在具体目标一中，我们建议研究人类 T1D 大鼠 BB 模型中的血清蛋白，目的是确定胰岛特异性蛋白在破坏后是否释放到循环中β细胞。使用信息学工具和传统的蛋白质测序方法，我们将识别与 β 细胞破坏和糖尿病相关的血清蛋白和肽。在具体目标二中，我们将研究纯化的人类胰岛的蛋白质组，目标是鉴定最常见的胰岛组织特异性蛋白质，特别关注鉴定其翻译后修饰的水平。与此同时，我们将开发一种基于 SELDI-TOF 的免疫测定芯片，该芯片由识别 β 细胞特异性、神经内分泌组织特异性和/或胰岛炎症相关蛋白标记物的抗体组成。标记的选择基于我们对丰富的胰岛转录本的了解以及我们从胰岛蛋白质组表征中获得的信息。确定了 Isl​​et 芯片的性能特征后，我们将返回大鼠模型，回顾性地寻找血清中的这些标记物，并确定它们与 β 细胞损伤和糖尿病的关联。我们组建了一个跨学科研究团队，其共同目标是开发内分泌胰腺β细胞破坏的标记物。

项目成果

AurkA inhibitors enhance the effects of B-RAF and MEK inhibitors in melanoma treatment.

• DOI: 10.1186/s12967-014-0216-z
• 发表时间: 2014-07-31
• 期刊: Journal of translational medicine
• 影响因子: 7.4
• 作者: Caputo E;Miceli R;Motti ML;Taté R;Fratangelo F;Botti G;Mozzillo N;Carriero MV;Cavalcanti E;Palmieri G;Ciliberto G;Pirozzi G;Ascierto PA
• 通讯作者: Ascierto PA