Development of a Blood-based Test for Identifying Synucleinopathy in Patients with Dementia

项目摘要 在此Fast Track SBIR应用程序中，NeuroDex，Inc.建议利用其在细胞外囊泡（EV）方面的专业知识， 免疫亲和分离，以开发一种血液测试，以高灵敏度检测突触核蛋白病， 帮助临床医生识别抗精神病药物有害和禁忌的患者。 开发和商业化一种用于突触核蛋白病的敏感血液检测将解决一个问题， 选择适当的抗精神病药物治疗以避免严重不良反应的主要未满足需求。 拟议的工作利用了NeuroDex开发的专有程序，用于使用 细胞特异性表面标志物的免疫亲和性。使用这种方法，该公司展示了一个> 20倍 NDE的信噪比增加，这是分析aSYN所必需的特异性程度，因为> 血浆中90%的aSYN不起源于大脑，与疾病无关。以前的努力衡量 未处理血浆中的aSYN没有产生任何诊断价值，但在NDE分离后进行测量 提供强大的分类功能。利用这种方法，NeuroDex已经开发了检测aSYN的方法， 以及神经元、小胶质细胞和少突胶质细胞衍生的EV的表面。这些试验成功地 区分35名健康对照，51名PD患者和30名MSA患者具有高准确性。现在用NeuroDex 建议完成发现阶段（第一阶段），并进行分析和临床验证（第二阶段）。 第一阶段-目标1。确定要纳入突触核蛋白病检测组的最终生物标志物。使用 195个样本来自健康个体和一系列尸检证实的突触核蛋白病患者，我们将 评估可能的生物标志物的不同组的性能，以选择具有最佳生物标志物的组合。 性能成功指标：≥ 80% PPV用于识别突触核蛋白病患者和≥ 90% NPV。目的 2.鉴定所选测定。成功验证：精密度CoV ≤ 20%，在8倍稀释范围内呈线性， 和LLQ与临床样品相容。进展至II期的Go/No Go标准：完成 在发现阶段，在一组生物标志物中包括一组定义的生物标志物，a）提供≥ 90%的NPV，和B）可以 以稳健的方式测量（精密度和再现性的可变性<20%）。第二阶段-目标1。分析 诊断分析的验证。里程碑和成功案例：1）成功审核CLIA实验室SOP，2） FDA突破性认定申报的分析数据包。目标2.初步临床验证 在CLIA认证的实验室进行诊断检测。我们将用700份来自患者的样本验证检测方法 痴呆伴或不伴共核蛋白病的患者和年龄匹配的对照组。成就与成功： 主要：NPV ≥ 90%;次要：PPV ≥ 80%。影响-新型血液的开发和严格验证 用于鉴别诊断具有突触核蛋白病理学痴呆的测试将产生商业上可行的 诊断，以告知适当的治疗计划和临床试验分层，以改善药物开发。

PROJECT SUMMARY In this Fast Track SBIR application, NeuroDex, Inc. proposes to use its expertise in extracellular vesicle (EV) immunoaffinity isolation to develop a blood test that detects synucleinopathies with high sensitivity, with the long- term goal of helping clinicians identify patients for whom antipsychotic drugs are harmful and contraindicated. Development and commercialization of a sensitive blood test for synucleinopathies would address a major unmet need for selecting appropriate antipsychotic treatments that avoid severe adverse effect. The proposed work leverages a proprietary procedure NeuroDex developed for isolating EVs from plasma using immunoaffinity for cell-specific surface markers. Using this approach, the company demonstrated a > 20-fold increase in signal to noise ratio of NDEs, a degree of specificity that is essential for analyzing aSYN because > 90% of aSYN in plasma does not originate in the brain and is not disease related. Previous efforts to measure aSYN in unprocessed plasma have not yielded any diagnostic value, but measurement after NDE isolation provides powerful classification. Leveraging this approach, NeuroDex has developed assays to test aSYN within and on the surface of neuron-, microglia-, and oligodendrocyte-derived EVs. These assays successfully distinguished 35 healthy controls, 51 PD patients, and 30 MSA patients with high accuracy. NeuroDex now proposes to complete the discovery phase (Phase I) and conduct analytical and clinical validation (Phase II). PHASE I—Aim 1. Identify final biomarkers to be included in the synucleinopathy detection panel. Using 195 samples from healthy individuals and patients with an array of autopsy-confirmed synucleinopathies, we will assess the performance of different panels of the possible biomarkers to select the combination with the best performance. Success Metric: ≥ 80% PPV for identifying patients with synucleinopathies and ≥ 90% NPV. Aim 2. Qualify the selected assays. Success Metrics: precision CoV ≤ 20%, linearity across 8-fold dilution range, and LLQ compatible with clinical samples. Go/No Go Criterion for Progression to Phase II: Completion of the discovery phase to include a defined set of biomarkers in a panel that a) provides ≥ 90% NPV and b) can be measured in a robust manner (variability in precision and reproducibility <20%). PHASE II—Aim 1. Analytical validation of the diagnostic assay. Milestones & Success Metrics: 1) Successful audits for CLIA lab SOP, 2) analytical data package for FDA breakthrough designation submission. Aim 2. Preliminary clinical validation of the diagnostic assay in a CLIA-certified lab. We will validate the assays with 700 samples from patients with dementia with or without synucleinopathies and age-matched controls. Milestones & Success Metrics: primary: NPV ≥ 90%; secondary: PPV ≥ 80%. Impact—Development and rigorous validation of a novel blood test for differential diagnosis of dementia with synuclein pathology would produce a commercially-viable diagnostic to inform appropriate treatment plans and clinical trial stratification for improved drug development.