Imaging Protein Aggregates for Early Diagnosis and Monitoring of Parkinson's Disease

阿尔茨海默氏症和帕金森氏症等神经生殖疾病是主要的健康问题，随着我们寿命的延长，这些疾病的负担正在增加，英国每年在医疗和社会护理、非正式护理费用和生产力损失方面花费约300亿GB。研究和治疗这些疾病的一个重要限制因素是缺乏简单的诊断和监测测试。对于这个项目，我们将在我们已经获得的非常令人鼓舞的初步结果的基础上，将重点放在帕金森氏症上。我们的目标是开发一种早期诊断帕金森氏症的方法，这种方法可以在血液或唾液等容易获得的样本上进行，从而可以用于筛查处于高危人群中的人。如果这是可能的，它将使正在发展的疾病的患者在他们出现症状之前被识别出来，因此任何可用的治疗都可能更有效。我们还旨在通过比较从不同疾病阶段的患者那里收集的样本，以及随着时间的推移从同一个人那里收集的重复样本，来确定我们的新测试是否能够随着时间的推移发现疾病的变化。如果是这样的话，这将为临床试验中监测这种疾病提供一个亟需的标记物。在帕金森氏病和其他神经退行性疾病的发展过程中，蛋白质聚集在一起，在大脑中形成更大的团块。这些团块被分泌到洗澡大脑和脊髓的液体(脑脊液)中，并进入血液中。我们发现，在被诊断出患有这种疾病的1-2年内，血液中存在较大比例的称为a-突触核蛋白的较大蛋白质块。我们通过选择性地在表面捕捉这些团块，然后通过添加抗体使它们可见并使用灵敏的荧光显微镜对它们进行成像来检测这些团块。我们将以我们的初步数据为基础，开发一种自动执行这些实验的方法，这样我们就可以同时在数百个样本中检测血液中所有关键蛋白质的团块。然后，我们将进行测试，看看我们是否能找到聚合体中的变化，这些变化最能区分患有这种疾病的患者和没有这种疾病的健康“对照”。我们还将测试血液是否是最好的样本，或者唾液或鼻拭子单独或与血液一起使用是否能更好地区分帕金森氏症患者和对照组。我们将从一大群处于不同疾病阶段的帕金森氏症患者以及对照组中收集一组新的样本，以测试优化后的方法效果如何，以及它是否与疾病严重程度有关。我们还将在稍后的时间点对从相同个人收集的样本进行重复测试，看看它是否会随着时间的推移而变化。我们还将分析患有快速眼动睡眠行为障碍(RBD)的睡眠问题患者的血液样本，其中许多人可能会继续发展为帕金森氏症，看看我们的方法是否也可以识别那些转化为帕金森氏症的特别高风险的人。这将展示该方法在早期疾病诊断中的潜力。到项目结束时，我们希望开发出一种基于血液或唾液的早期检测和监测帕金森病的方法，并已在临床样本中得到验证。然后，这种方法可以用来筛查这种疾病的“风险”人群(例如，由于RBD，携带遗传风险变异，或有轻微的神经异常)，以确定他们是否处于帕金森氏症的早期阶段。然后，同样的方法可能适用于其他疾病，如阿尔茨海默氏症或创伤性脑损伤，如足球运动员反复头球时经历的疾病早期诊断。我们希望该方法也将有助于随着时间的推移监测疾病过程的变化，这对于监测临床试验中新疗法的有效性至关重要。

Neurogenerative diseases such as Alzheimer's and Parkinson's diseases are major health problems whose burden is increasing as we live for longer, and cost the UK about £30 billion per year in health and social care, informal care costs and productivity losses. An important limiting factor in studying and treating these disorders is the lack of simple tests for diagnosis and monitoring. For this project we will focus on Parkinson's disease, building on very encouraging preliminary results that we have obtained. We aim to develop a method for the early diagnosis of Parkinson's disease that can be performed on easily available samples such as blood or saliva and so could be used to screen people in 'at risk' groups. If this is possible it would allow patients who are developing disease to be identified before they develop symptoms and hence any available treatment is likely to be much more effective. We also aim to establish whether our new test can pick up change in the disease over time by comparing samples collected from patients at different disease stages, as well as repeated samples collected from the same individuals over time. If so, this would provide a much needed marker for monitoring the disease in clinical trials.During the development of Parkinson's disease and other neurodegenerative diseases, proteins clump together and form larger clumps in the brain. These clumps are secreted into the fluid which bathes the brain and spinal cord (cerebrospinal fluid) and into the blood. We have found that a higher proportion of larger clumps of a protein called a-synuclein are present in the blood of patients within 1-2 years of being diagnosed with the disease. We detect these clumps by capturing them selectively on a surface and then imaging them by adding antibodies to make them visible and using a sensitive fluorescence microscope. We will build on our preliminary data to develop an automated way to perform these experiments so we can detect clumps of all the key proteins in blood, in hundreds of samples at the same time. We will then test to see whether we can find the changes in the aggregates what best distinguish between patients with the disease and healthy 'controls' without the disease. We will also test whether blood is the best sample to use or if saliva or nasal swabs alone or in combination with blood provide better discrimination between people with Parkinson's disease and controls. We will collect a new set of samples from a large group Parkinson's patients at different disease stages, as well as controls, to test how well the optimised method works, and whether it is linked to disease severity. We will also repeat the test in samples collected from the same individuals at a later timepoint to see whether it changes over time. We will also analyse blood samples from patients with a sleep problem called REM Sleep Behaviour Disorder (RBD), many of whom are likely to go on to develop Parkinson's disease, to see whether our method also can identify those at particularly high risk of conversion to Parkinson's. This would demonstrate the potential of the method for early disease diagnosis. By the end of the project we hope to have developed a blood or saliva based method for early detection and monitoring of Parkinson's disease that has been validated on clinical samples. This could then be used to screen people who are 'at risk' of the disease (e.g. due to RBD, carrying genetic risk variants, or having subtle neurological abnormalities) to see whether they are in early stages of developing Parkinson's. The same method might then be applicable to other diseases such as Alzheimer's disease or traumatic brain injury, as experienced by footballers from heading footballs repetitively, for early disease diagnosis. We hope the method will also be useful to monitor changes in the disease process over time which is critically important for monitoring the effectiveness of new therapies in clinical trials.