Optimization and Validation of an Indicator Cell Assay for Blood-Based Diagnosis of Alzheimer's Disease

项目摘要。PreCyte正在开发指示细胞分析平台(ICAP)，作为一种广泛适用和 廉价的血液化验，可用于疾病的早期检测，疾病阶段分层， 对各种疾病的治疗的预后和预测反应。ICAP使用培养的、标准化的 细胞作为生物传感器，利用细胞对血清中存在的疾病信号做出反应的能力 精致的灵敏度，而不是传统的依赖于直接检测血液中分子的分析方法。 发展ICAP包括将培养的细胞暴露于正常或疾病受试者的血清中，测量 全局差异响应模式，并使用它来构建基于表达的可靠疾病分类器 少量的基因。部署ICAP只涉及测量分类器基因的表达 使用经济高效的工具的功能。我们已经演示了ICAP用于基于血液的肺癌检测 以及在临床前和早期症状阶段检测阿尔茨海默病(AD)。 在SBIR第二阶段研究中，与传染病中心系统生物学研究所合作进行 疾病研究所和西雅图生物医学和临床研究所，PreCyte开发了一种ICAP- AD能够在多个队列中检测到临床前和早期症状阶段的AD。对于这个阶段IIb， 我们建议开发和验证临床ICAP AD，这是ICAP的高通量和廉价版本 用于快速选择AD疗法临床试验的受试者。从简单的血液测试，临床AD ICAP将 能够从60岁以上的受试者中进行选择，这些受试者要么是临床前期的，要么是早期的 有症状的阿尔茨海默病和非阿尔茨海默病的特异性，并可能具有额外的识别能力 这些候选人中的一部分将在2年内进入AD。临床测试系统的开发和验证 ICAP-AD将在四个具体目标下进行：1)开发临床ICAP-AD，2)建立关键分析的限值 验证分析稳健性的参数，3)验证临床ICAP的分析特征，以及4) 验证临床ICAP的诊断特征。本提案的目标是降低使用 一种基于细胞的检测方法，并为选择AD临床试验的受试者建立一个可靠且有效的平台 和耐心的诊断。在CLIA环境中实施后，此工具将显著降低成本并 通过快速选择早期受试者缩短AD治疗的临床试验持续时间，有可能 确定哪些人在发展为AD的2年内。它还将提高开发有效的 治疗阿尔茨海默病，因为它使患者能够针对疾病的临床前和早期症状阶段， 它们已被证明比后期更容易治疗，也可以用于监测 AD进展和对治疗的反应。

Project Summary. PreCyte is developing the Indicator Cell Assay Platform (iCAP) as a broadly applicable and inexpensive blood-based assay that can be used for the early detection of disease, disease stage stratification, prognosis and predicting response to therapy for a variety of diseases. The iCAP uses cultured, standardized cells as biosensors, capitalizing on the ability of cells to respond to disease signals present in serum with exquisite sensitivity, as opposed to traditional assays that rely on direct detection of molecules in blood. Developing the iCAP involves exposing cultured cells to serum from normal or diseased subjects, measuring a global differential response pattern, and using it to build a reliable disease classifier based on the expression of a small number of genes. Deploying the iCAP involves measuring only expression of genes that are classifier features using cost-effective tools. We have demonstrated the iCAP for blood-based detection of lung cancer and detection of Alzheimer's disease (AD) at preclinical and early symptomatic stages. In the SBIR Phase II study, conducted in collaboration with the Institute for Systems Biology, Center for Infectious Disease Research, and Seattle institute of Biomedical and Clinical Research, PreCyte has developed an iCAP- AD capable of detecting AD at preclinical and early symptomatic stages in multiple cohorts. For this Phase IIb, we propose to develop and validate a clinical iCAP AD, a high-throughput and inexpensive version of the iCAP for rapidly selecting subjects for clinical trials of AD therapies. From a simple blood test, the clinical AD iCAP will be able to select from a population of subjects 60 years and older, those with either preclinical or early symptomatic AD and specificity against those with non-AD dementia, and may have additional capacity to identify a subset of these candidates who will progress to AD within 2 years. Development and validation of the clinical iCAP-AD will be done in four specific aims: 1) Develop the clinical iCAP-AD, 2) Establish limits for key assay parameters to demonstrate assay robustness, 3) Validate analytical characteristics of the clinical iCAP, and 4) Validate diagnostic characteristics of the clinical iCAP. The goal of this proposal is to mitigate the risks of using a cell-based assay and establish a robust and validated platform for use in selecting subjects for AD clinical trials and patient diagnosis. After implementation in a CLIA environment, this tool will significantly reduce cost and shorten the duration of clinical trials for AD treatments by rapidly selecting early-stage subjects, with potential to identify those who are within 2 years of developing AD. It will also improve success rate of developing an effective treatment for AD as it enables the targeting of patients at preclinical and early symptomatic stages of the disease, which have been demonstrated to be more treatable than later stages, and could also be used for monitoring AD progression and response to treatment.