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系统疾病是主要的健康问题，随着我们寿命的延长，其负担也在增加，英国每年在健康和社会护理、非正式护理成本和生产力损失方面的成本约为300亿英镑。研究和治疗这些疾病的一个重要限制因素是缺乏诊断和监测的简单测试。在这个项目中，我们将把重点放在帕金森病上，以我们已经获得的非常令人鼓舞的初步结果为基础。我们的目标是开发一种早期诊断帕金森氏症的方法，这种方法可以在容易获得的血液或唾液样本上进行，因此可以用于筛查“高危”人群。如果这是可能的，它将使正在发病的患者在出现症状之前被识别出来，因此任何现有的治疗方法都可能更加有效。我们还旨在通过比较从不同疾病阶段的患者收集的样本，以及从同一个体收集的重复样本，确定我们的新测试是否可以随着时间的推移而发现疾病的变化。如果是这样，这将为临床试验中监测该疾病提供一个急需的标记物。在帕金森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。这些团块被分泌到沐浴大脑和脊髓的液体（脑脊液）和血液中。我们发现，在被诊断患有这种疾病的1-2年内，患者血液中存在更高比例的称为a-突触核蛋白的较大蛋白质团块。我们通过选择性地在表面捕获这些团块来检测它们，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。我们将在初步数据的基础上开发一种自动化的方法来进行这些实验，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总量的变化，从而最好地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使用或与血液结合使用是否能更好地区分帕金森病患者和对照组。我们将从一大群处于不同疾病阶段的帕金森患者以及对照组中收集一组新的样本，以测试优化方法的效果如何，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从同一个人收集的样本中重复测试，以查看它是否随时间变化。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，其中许多人可能会发展为帕金森病，看看我们的方法是否也能识别出那些转化为帕金森病的风险特别高的人。这将证明该方法在早期疾病诊断方面的潜力。到项目结束时，我们希望开发出一种基于血液或唾液的方法，用于帕金森病的早期检测和监测，并已在临床样本中得到验证。然后，这可以用来筛选有患帕金森病风险的人（例如，由于RBD，携带遗传风险变异，或有细微的神经异常），看看他们是否处于帕金森病的早期阶段。同样的方法可能适用于其他疾病，如阿尔茨海默病或创伤性脑损伤，如足球运动员因反复头球而经历的早期疾病诊断。我们希望该方法也能用于监测疾病过程随时间的变化，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.