Quantitative analysis of key protein phosphorylation events as pathway biomarkers of metabolic disease in human cells.

久坐不动的生活方式和富含脂肪的饮食相结合，导致了与代谢功能障碍相关的健康问题的激增。例如，英国有230万人患有2型糖尿病，而十分之一的育龄妇女会患有多囊卵巢综合征。事实上，仅仅是超重就会增加患这些疾病的几率。仅2型糖尿病一项每年就花费NHS约20亿英镑，而糖尿病诊断预计在未来10年内将翻一番。2型糖尿病是心脏病、失明、截肢和中风的主要原因，因此会降低个人多年的生活质量，并使预期寿命缩短约10年。2型糖尿病的主要特征之一是对胰岛素反应差。所有证据都指向这个？胰岛素抵抗?发生在2型糖尿病发展之前，但目前在技术上不可能筛查一般人群的胰岛素抵抗。我们坚信，更早的诊断和更有针对性的治疗将防止2型糖尿病的进展，从而防止该疾病的破坏性并发症。此外，胰岛素抵抗与所有肥胖相关疾病密切相关，包括多囊卵巢综合征、高血压和脂肪肝疾病。因此，相同的诊断方法和潜在的相同的治疗方法可以应用于所有这些？代谢疾病?。在本提案中，我们的目标是开发新的方法（使用最先进的质谱技术）来准确测量细胞内胰岛素作用的多种生物标志物。胰岛素诱导细胞内许多蛋白质的修饰，当细胞或组织具有胰岛素抵抗性时，这些修饰不会以相同的模式发生。因此，这些生物标记物的精确测量给出了三个结果；首先，暴露于胰岛素后的修饰模式提供了细胞或组织的胰岛素敏感性，其次，异常模式提供了细胞或组织中分子缺陷的具体位置，最后，分子缺陷的识别允许对不同形式的胰岛素抵抗进行分类，以便确定对该亚组的正确治疗。该项目将分为两个阶段，方法的开发和新方法的验证，使用细胞和仔细收集的肌肉活检，这些活检来自不同体脂量（肥胖）的志愿者，因此有一系列不同水平的胰岛素抵抗。

A combination of a more sedentary lifestyle and diets rich in fat has resulted in an explosion of health problems associated with metabolic dysfunction. For example, there are 2.3 million people in the UK with Type 2 diabetes, while one in ten women of child bearing age will experience polycystic ovarian syndrome. Indeed, simply being overweight increases the chances of developing these diseases. Type 2 diabetes alone costs the NHS around #2 billion pounds per year, while diabetes diagnosis is expected to double in the next 10 years. Type 2 diabetes is a major cause of heart disease, blindness, amputation and stroke, so reduces quality of life for individuals for many years as well as reducing life expectancy by around 10 years. One of the key features of Type 2 diabetes is poor response to the hormone insulin. All evidence points to this ?insulin resistance? occurring prior to the development of Type 2 diabetes, but it is technically impossible at present to screen for insulin resistance in the general population. We strongly believe that earlier diagnosis and more targeted therapy would prevent progression to Type 2 diabetes, hence preventing the devastating complications of the disease. In addition, insulin resistance is strongly associated with all obesity related diseases, including polycystic ovarian syndrome, hypertension and fatty liver disease. Therefore the same diagnostic approach, and potentially the same therapy could be applied to all these ?metabolic diseases?.In this proposal we aim to develop novel methodology (using state-of the-art Mass Spectrometry techniques) to accurately measure multiple biomarkers of insulin action inside cells. Insulin induces modifications of many proteins inside cells, and these modifications do not occur in the same pattern when cells or tissues are insulin resistant. Therefore accurate measurement of these biomarkers gives three things; firstly the pattern of modification after exposure to insulin provides the insulin sensitivity of the cells or tissues, secondly, an abnormal pattern provides the specific location of the molecular defect in the cells or tissues, and finally identification of the molecular defect allows classification of different forms of insulin resistance in order to identify the correct treatment for that subgroup.The project will be split into two phases, the development of the methodology, and validation of the new methodology using cells and carefully collected muscle biopsies from volunteers with different amounts of body fat (obesity), and therefore a range of different levels of insulin resistance.