Diagnostic aptamer reagents to develop multi-analyte blood test for pre-clinical, mild and moderate Alzheimer's disease

诊断适配子试剂开发用于临床前、轻度和 中度阿尔茨海默病 Aptus Biosciences，LLC 巴拉特·加万德 摘要 今天，620万美国人患有阿尔茨海默病(AD)，每年造成3550亿美元的损失 医疗费用。随着美国65岁以上人口的增长，预计患有AD的美国人数量将会增加 到2050年增加到1270万。AD是一种进展缓慢的疾病，可能需要长达20年的时间才能 症状是可以辨认的。虽然新的治疗方法正在涌现，但这可能有助于控制或改变这种疾病， 使用简单的血液测试进行早期检测对AD患者至关重要。重要的是，最近的研究表明 AD特异的血液生物标记物取得了令人振奋的结果。然而，显然缺乏高 可以与这些生物标志物结合并用于开发敏感和易于获取的血液的亲和试剂 测试。 阿普图斯生物科学公司将使用改良的适体选择技术，创造高度敏感和特异的 检测阿尔茨海默病血液生物标志物的试剂，这些生物标志物会导致神经原纤维缠结(NFT)，这是AD的一个标志。我们 将提供一种使用疏水的临时体外选择方法开发的适体试剂 可与tau蛋白结合的修饰核苷酸和多种形式的磷酸化tau(p-tau)蛋白 具有皮摩尔亲和力。每个靶向选择将产生与特定的不同表位结合的适体 具有皮摩尔亲和力的p-tau靶标，将用于开发简单的珠基适配子三明治 一个适配子捕获p-tau生物标记物和其他适配子的检测将导致信号产生 测量这些分析物的血浆浓度。我们的改良适体试剂有可能比 未修饰的适体和大的基于抗体的试剂，它们不那么敏感或特异，因为它们的 多分析物分析中的大尺寸、低亲和力和交叉反应。具体来说，我们将使用选择来创建 修饰的适配子，结合特定的有效磷酸化的tau亚型。此外，我们还将开发一种 简易珠基多分析适体夹心法测定血浆样品中的p-tau异构体 从脑脊液中提取β阳性的AD确诊患者42例，并与血浆样本进行比较 从认知正常、Aβ阴性的42例无AD的个体中获得。 如果第一阶段的资助成功，在第二阶段，我们将优化适体试剂以提高灵敏度和 分析的特异性，放大适体试剂的生产，使用训练开发基于云的算法 样本集，并使用测试样本验证分析，以正确识别大型队列中的早期阿尔茨海默病。多种多样- 基于分析适体的分析将使其远远优于现有的定性、侵入性和昂贵的诊断 测试。如果成功，这一简单的血液测试将改善数百万处于高风险的美国人的生活 由于测试具有早期发现的能力而发展为阿尔茨海默病，从而导致早期治疗干预 以及减轻医疗体系的巨大经济负担。

Diagnostic aptamer reagents to develop multi-analyte blood test for pre-clinical, mild and moderate Alzheimer’s disease Aptus Biosciences, LLC Bharat Gawande Summary Today, 6.2 million Americans are living with Alzheimer’s disease (AD), resulting in $355 billion in annual medical care costs. As the US population over age 65 grows, the number of Americans living with AD is expected to increase to 12.7 million by 2050. AD is a slow progressing disease, and it may take up to 20 years before symptoms are recognizable. While new treatments are emerging, that may help to control or modify the disease, early detection using a simple blood test is critical to individuals with AD. Importantly, recent studies have shown promising results with blood-based biomarkers that are specific for AD. However, there is clearly a lack of high affinity reagents that can bind to these biomarkers and be used to develop sensitive and easily accessible blood tests. Aptus Biosciences will use modified aptamer selection technology, to create highly sensitive and specific reagents that detect blood biomarkers of AD which cause neurofibrillary tangles (NFTs), a hallmark of AD. We will deliver aptamer reagents developed from an improvised In Vitro selection method using hydrophobic modified nucleotides that can bind to tau protein, and multiple isoforms of phosphorylated tau (p-tau) protein with pico-molar affinity. Each targeted selection will generate aptamers that bind to various epitopes on specific p-tau targets with picomolar affinity and that will be used to develop a simple bead-based aptamer sandwich assay in which one aptamer will capture p-tau biomarker and other aptamer will result in a signal generation to measure the plasma concentrations of these analytes. Our modified aptamer reagents are potentially better than unmodified aptamers and large antibody-based reagents, which are not as sensitive or specific because of their large size, low affinity, and cross-reactivity in multi-analyte assays. Specifically, we will use selections to create modified aptamers that bind to specific validated phosphorylated isoforms of tau. In addition, we will develop a simple bead-based multi-analyte aptamer sandwich assay to measure p-tau isoforms in plasma samples obtained from cerebrospinal fluid (CSF) Aβ42 positive, AD-confirmed individuals and compare them with plasma samples obtained from cognitively normal, Aβ42 negative individuals without AD. If this Phase I grant is successful, in Phase II, we will optimize aptamer reagents to improve the sensitivity and specificity of the assay, scale-up aptamer reagent production, develop a cloud-based algorithm using a training sample set, and validate the assay using test samples to correctly identify early AD in a large cohort. The multi- analyte aptamer-based assay will make it far superior to existing qualitative, invasive, and expensive diagnostic tests. If successful, this simple blood test will enhance the lives of millions of Americans who are at high risk of developing AD due to the ability of the test for early detection, thus resulting in early treatment interventions and decreasing the huge economic burden to the healthcare system.