Markers for T2DM, Risk of T2DM & Aptamer Array Diagnosis

描述(由申请人提供)：该项目的直接目标是识别用于2型糖尿病的新的生物标记蛋白和早期检测2型糖尿病的风险，并测试一种新的光适配子阵列标记分析系统。更长期的目标是在一个强大和可靠的光适配子阵列诊断平台中培养和验证2型糖尿病风险的新标记物(以及其他人发现的标记物)的测量。这项工作背后的假设是，对蛋白质标记物的深入全球搜索将揭示多个标记物的模式，这些标记物将比目前的诊断方法更早、更可靠地发出2型糖尿病(和其他疾病)风险状态的信号，而不需要葡萄糖注射和广泛的多点耐量测量等干扰。全球蛋白质组学方法非常适合于疾病标志物的发现，但对于直接诊断来说似乎过于昂贵，而光适配子系统似乎非常适合提供常规、可靠、高通量的诊断。 我们计划使用蒙大拿州立大学开发的新型超灵敏多重荧光检测染料，以液态IEF预分离的2D凝胶格式，更深入地研究血清蛋白质组中的生物标志物和生物标志物亚型。这种改进的方法提供了一种强大的手段来检测样品之间蛋白质表达的差异，尤其是在测量修饰蛋白质异构体的变化方面。我们将补充和比较2D凝胶分析和高分辨率2D-LC/MS/MS在相同组分上的分析，使用同位素半胱氨酸和赖氨酸标记来测量蛋白质和肽表达的变化。伍斯特大学大众医学中心的一个临床小组(该小组正在寻求为2型糖尿病风险人群开发更有效的生活方式改变策略)正在提供具有良好特征的患者和对照组的样本。该项目将我们正在开发和应用新的靶向蛋白质组检测技术的学术团队与来自行业的优秀研究人员团队聚集在一起，他们贡献了先进的技术资源和强大的诊断平台。 与公共健康相关：在美国，大约有500万人被认为患有未确诊的2型糖尿病，而且这个数字似乎还在增长。在诊断为2型糖尿病时，许多患者已经有严重的糖尿病并发症，并极大地增加了患心脏病的风险。现有的筛查方法，如口服葡萄糖耐量试验似乎效果不大，价格昂贵，难以实施，而且几乎没有证据表明，利用目前的技术筛查没有症状的成年人患有未诊断的糖尿病。该项目旨在确定诊断目标，并促进开发更简便、廉价、敏感和可靠的临床测试。

DESCRIPTION (provided by applicant): The immediate goal of this project is to identify new biomarker proteins for type 2 diabetes and early detection of risk of type 2 diabetes and to test a new photoaptamer array marker analysis system. The longer term goal is to foster and validate the measurement of new markers for the risk of type 2 diabetes (and markers found by others) in a robust and reliable photoaptamer array diagnostic platform. The hypothesis underlying this work is that a deep, global search for protein markers will reveal patterns of multiple markers that will signal type 2 diabetes (and other disease) risk states significantly earlier and much more reliably than current diagnostics, without requiring perturbations such as glucose administration and extensive multipoint tolerance measures. Global proteomics methods are well suited for disease marker discovery but appear to be too expensive for direct diagnosis, while the photoaptamer systems appear to be extremely well suited for providing routine, reliable, high-throughput diagnosis. We plan to search more deeply into the blood serum proteome for biomarkers and biomarker isoforms than has been accomplished previously, using new ultrasensitive multiplex fluorescent detection dyes developed at Montana State University-in a 2D gel format with liquid state IEF prefractionation. This improved methodology provides a powerful means to detect differences in expression of proteins between samples and is especially powerful for measuring changes in modified protein isoforms. We will complement and compare the 2D gel analysis with high resolution 2D-LC/MS/MS on the same fractions, using isotopic cysteine and lysine tags for measuring changes in protein and peptide expression. A clinical group at the U Mass Medical Center, Worcester (that is seeking to develop more effective lifestyle modification strategies for people at risk of type 2 diabetes) is providing samples from well characterized patients and controls. This project brings together our academic group that is developing and applying new targeted proteomics detection technologies with outstanding groups of investigators from industry that are contributing advanced technology resources and a powerful diagnostic platform. Relevance to public health: Approximately 5 million people in the US are thought to have undiagnosed Type 2 diabetes and the numbers appear to be growing. At the time of diagnosis of Type 2 diabetes many patients already have severe complications of diabetes and are a greatly increased risk of heart disease. Available screening methods, such as the oral glucose tolerance test appear to be of modest effectiveness, are expensive and difficult to carry out and there is little evidence of benefit from screening asymptomatic adults for undiagnosed diabetes with current technology. This project aims to identify diagostic targets and to foster the development more facile, inexpensive, sensitive, and reliable clinical tests.