An Ultrasensitive, High-Throughput, and Low-Cost Autoimmunity Panel for the Early Diagnosis and Improved Treatment of Type 1 Diabetes

1型糖尿病(T1D)是一种破坏性的自身免疫性疾病，会破坏胰岛细胞胰岛素的产生。数以百万计的美国人，特别是儿童，受到影响，必须依赖终身胰岛素注射，并与危及生命的并发症作斗争。大多数新的T1D病例是在紧急情况下诊断出来的，表现为潜在的致命性糖尿病酮症酸中毒(DKA)。目前能够对T1D采取预防措施的临床前诊断测试速度很慢，而且在三级护理中心之外不易获得。在这项提议中，Enable Biosciences将进一步开发和验证一种新的低成本和高通量的免疫分析方法，以快速和灵敏地筛选预测1型糖尿病风险的四种主要自身抗体标记。这样的测试可以取代目前昂贵、缓慢的放射性方法。通过降低成本，同时利用强大的极低样本量分析保持灵敏度和特异性，Enable Bioscience的产品将允许增加检测，以在干预的最佳时间揭示普通人群中早期的T1D。在第一阶段资助期间，Enable使用其核心抗体检测凝集-聚合酶链式反应(ADAP)技术制作了一个由3名成员组成的T1D自身抗体小组的原型，并在消除对放射性试剂的依赖的同时，证明了与黄金标准测试的基本等价性。在这份第二阶段的提案中，Enable提出了一个全面的计划，通过使用专用的机器人液体处理系统实现自动化来增强该仪表板，从而在进一步降低成本的同时极大地提高分析吞吐量和可靠性。我们还将调整我们的分析化学，使其能够使用Taqman探针。这将进一步加速吞吐量并降低成本。在第一阶段工作的初步结果的基础上，我们将整合第四种也是最后一种T1D自身抗体(ZnT8)，创建世界上第一个“4合1”筛查小组，创造一种为T1D筛查和诊断提供最高水平风险评估的产品。这些经过优化的自动化仪表板将通过我们广泛的热心学术和机构合作伙伴网络提供的数千个临床样本进行严格验证，这些合作伙伴包括Joslin糖尿病中心、斯坦福医学院、T1D交易所和塔夫茨儿童医院。这些数据将用于规划和指导商业化工作，包括从提供CLIA认证的实验室服务到FDA批准的试剂盒的分阶段方法，以及配套的集成样本到答案的硬件设备，以便在世界各地的研究和临床实验室和筛查地点轻松、低技能地使用和部署。

Type 1 diabetes (T1D) is a devastating autoimmune disease that destroys islet-cell insulin production. Millions of Americans, especially children, are affected and must rely on lifelong insulin administration and struggle with life-threatening co-morbidities. Most new cases of T1D are diagnosed in emergency settings with presentation in potentially deadly diabetic ketoacidosis (DKA). Current pre-clinical diagnostic tests that can permit preventive action for T1D are slow and not readily available outside of tertiary care centers. In this proposal Enable Biosciences will further develop and validate a novel low-cost and high-throughput immunoassay to rapidly and sensitively screen for the four cardinal autoantibody markers that predict risk for type 1 diabetes. Such a test could supplant the current expensive, slow radioactive method. By reducing costs while retaining sensitivity and specificity with a robust very low-sample volume assay, Enable Biosciences' product will permit increased testing to reveal incipient T1D in the general population at the optimal time for intervention. During the Phase I funding period, Enable prototyped a 3-member T1D autoantibody panel using their core Antibody Detection by Agglutination-PCR (ADAP) technology and demonstrated substantial equivalence with the gold-standard assay while removing reliance on radioactive reagents. In this Phase 2 proposal, Enable presents a comprehensive plan to enhance this panel via automation using a dedicated robotic liquid handling system, greatly increasing assay throughput and reliability while further reducing costs. We will also adapt our assay chemistry to be able to use Taqman probes. This will result in further throughput acceleration and cost reductions. Building upon promising preliminary results from Phase 1 work, we will incorporate the fourth and final T1D autoantibody (ZnT8) to create the world's first ever “4-in-1” screening panel, creating a product that provides the highest level of risk assessment for T1D screening and diagnosis. These optimized and automated panels will then be rigorously validated with thousands of clinical samples provided by our wide network of enthusiastic academic and institutional partners that include the Joslin Diabetes Center, Stanford School of Medicine, the T1D Exchange and Tufts Children's Hospital. This data will be used to plan and direct commercialization efforts that will include a phased approach from delivering a CLIA-certified laboratory service to FDA-approved reagent kits with companion integrated sample-to-answer hardware devices for easy, low-skill use and deployment across research and clinical laboratories and screening sites worldwide.