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（T1 D）是一种破坏胰岛细胞胰岛素产生的破坏性自身免疫性疾病。数以百万计的美国人，特别是儿童，受到影响，必须依靠终身胰岛素管理和与危及生命的合并症作斗争。大多数新发T1 D病例是在紧急情况下诊断的，表现为潜在致命的糖尿病酮症酸中毒（DKA）。目前的临床前诊断测试，可以允许T1 D的预防措施是缓慢的，并不容易获得三级保健中心以外。在这项提案中，Enable Biosciences将进一步开发和验证一种新型的低成本和高通量免疫测定法，以快速灵敏地筛选预测1型糖尿病风险的四种主要自身抗体标志物。这样的测试可以取代目前昂贵的，缓慢的放射性方法。通过降低成本，同时保持灵敏度和特异性与一个强大的非常低的样本量测定，启用生物科学公司的产品将允许增加测试，以揭示早期T1 D在一般人群中的最佳干预时间。在I期资助期间，Enable使用其核心抗体检测-聚合酶链反应（ADAP）技术制作了3人T1 D自身抗体样本组原型，并证明了与金标准检测试剂盒的实质等同性，同时消除了对放射性试剂的依赖。在第二阶段提案中，Enable提出了一项全面的计划，通过使用专用机器人液体处理系统的自动化来增强该面板，大大提高了分析吞吐量和可靠性，同时进一步降低了成本。我们还将调整我们的测定化学，以便能够使用Taqman探针。这将导致进一步的吞吐量加速和成本降低。在第一阶段工作的初步结果的基础上，我们将结合第四种也是最后一种T1 D自身抗体（ZnT 8）来创建世界上第一个“4合1”筛查面板，创建一种为T1 D筛查和诊断提供最高水平风险评估的产品。这些优化和自动化的面板将通过我们广泛的学术和机构合作伙伴网络提供的数千份临床样本进行严格验证，这些合作伙伴包括Joslin糖尿病中心，斯坦福大学医学院，T1 D交换和塔夫茨儿童医院。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硬件设备，以便在全球研究和临床实验室以及筛查中心轻松，低技能使用和部署。

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.