Novel high-throughput approaches for the rapid analysis of target engagement by mass spectrometry

阿尔茨海默病（AD）是当今已知的最常见的神经退行性疾病之一，影响着全世界约5000万人。致病机制的特点是形成有毒的a - β寡聚物和聚集体，破坏神经元功能。受a - β影响最大的神经细胞类型之一是谷氨酸能神经元，它需要接收和传递感觉信号。例如，嗅觉丧失（嗅觉功能障碍）是阿尔茨海默病人类最早的症状之一。同样，在秀丽隐杆线虫中，通过谷氨酸能神经元介导的行为化学感觉缺陷发生在神经系统中表达a - β的幼龄动物身上。然而，我们不知道在AD进展过程中，哪种谷氨酸能神经元亚型由于a- β相关毒性首先失去信号活性，也不知道如何保护神经元功能。该项目将确定在AD进展过程中哪个神经元回路首先屈服于a - β表达，以及Hsp90如何保护谷氨酸能神经元功能。因此，我们将建立一种改进的秀丽隐杆线虫a- β相关阿尔茨海默病（AD）模型，该模型可以监测整个衰老和疾病进展过程中的神经元信号活动；结合钙报告和谷氨酸传感器。报告器将允许确定何时神经元信号活动下降，哪些神经元亚型首先受到影响，以及保护分子（如伴侣和药理学化合物）是否保护神经元信号活动。确定受a - β表达影响的确切神经元亚型以及Hsp90如何保留信号功能将为阿尔茨海默病的发展提供新的见解。这将有助于开发新的治疗方案，可以在疾病过程的早期应用，以延迟甚至防止疾病的进一步进展。改良的秀丽隐杆线虫AD报告基因将取代小鼠AD等神经退行性疾病模型，在神经元信号活动水平和与行为输出相关的水平上研究伴侣和药理化合物的保护作用。

Alzheimer's disease (AD) is one of the most common form of neurodegenerative diseases known today, affecting approximately 50 million people worldwide. The pathogenic mechanism is characterised by the formation of toxic A-beta oligomeric species and aggregates that disrupt neuronal function. One of the neuronal cell types most affected by A-beta are glutamatergic neurons, which are required to receive and transmit sensory signals. For example, loss of the sense of smell (olfactory dysfunction) is one of the earliest symptoms of AD in humans. Similarly, in C. elegans, chemosensory deficits in behaviour mediated via glutamatergic neurons, occurs in already young animals expressing A-beta in the nervous system. We however do not know which glutamatergic neuronal subtype loses its signalling activity first as a consequence of A-beta-associated toxicity during AD progression and how neuronal function can be protected.This project will determine which neuronal circuit succumbs first to A-beta expression during AD progression and how Hsp90 protects glutamatergic neuronal function. Therefore, we will establish an improved C. elegans model of A-beta-associated Alzheimer's Disease (AD) that allows to monitor neuronal signalling activity throughout aging and progression of the disease; using a calcium reporter in combination with a glutamate sensor. The reporter would allow to determine when neuronal signalling activity declines, which neuronal subtypes are affected first and whether protective molecules (such as chaperones and pharmacological compounds) safeguard neuronal signalling activity. Identification of the exact neuronal subtype affected by A-beta expression and how Hsp90 preserves signalling function will provide novel insights into how AD develops. This will facilitate the development of new treatment options that can be applied early in the disease process to delay or even prevent further progression of the disease. The improved C. elegans AD reporter will replace mouse neurodegenerative disease models such as AD, to investigate the protective effects of chaperones and pharmacological compounds at the level of neuronal signalling activity and correlated to behavioural output.