Markers for T2DM, Risk of T2DM & Aptamer Array Diagnosis

描述（由申请人提供）：该项目的近期目标是鉴定2型糖尿病的新生物标志物蛋白和2型糖尿病风险的早期检测，并测试新的光适体阵列标记物分析系统。长期目标是在一个强大可靠的光适体阵列诊断平台中促进和验证2型糖尿病风险的新标志物（以及其他人发现的标志物）的测量。这项工作背后的假设是，对蛋白质标记物的深入、全球搜索将揭示多种标记物的模式，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，而不需要葡萄糖给药和广泛的多点耐受性测量等干扰。全局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，而光适体系统似乎非常适合于提供常规的、可靠的、高通量的诊断。 我们计划搜索更深入的血清蛋白质组的生物标志物和生物标志物亚型比以前已经完成，使用新的超灵敏的多重荧光检测染料在蒙大拿州立大学开发的二维凝胶格式与液态IEF prefractionation。这种改进的方法提供了检测样品之间蛋白质表达差异的有力手段，并且对于测量经修饰的蛋白质同种型的变化特别有力。我们将使用同位素半胱氨酸和赖氨酸标签测量蛋白质和肽表达的变化，对相同组分的2D凝胶分析与高分辨率2D-LC/MS/MS进行补充和比较。伍斯特大学医学中心的一个临床小组（正在寻求为有2型糖尿病风险的人制定更有效的生活方式改变策略）正在提供来自特征良好的患者和对照组的样本。该项目汇集了我们正在开发和应用新的靶向蛋白质组学检测技术的学术团队，以及来自行业的优秀研究人员团队，他们正在贡献先进的技术资源和强大的诊断平台。 与公共卫生的相关性：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2型糖尿病时，许多患者已经患有糖尿病的严重并发症，并且患心脏病的风险大大增加。现有的筛查方法，如口服葡萄糖耐量试验，似乎是适度的有效性，是昂贵的，难以进行，几乎没有证据表明，从筛查无症状的成年人未确诊的糖尿病与目前的技术的好处。该项目旨在确定诊断靶点，并促进开发更简便，廉价，灵敏和可靠的临床试验。

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.