Multiplexed DNA Origami-Based Biomarker Detection Assay for Early Dx of Arthritis

基于多重 DNA 折纸的生物标志物检测分析用于关节炎早期 Dx

• 批准号: 8523477
• 负责人: JOHN E MUELLER
• 金额: $ 24.88万
• 依托单位: LYNNTECH, INC.
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2014-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8523477
• 关键词: Adult; Affect; Affinity; American; Antibodies; Area; Arthritis; Binding; Binding Sites; Biological Assay; Biological Markers; Body Weight decreased; Cartilage; Centers for Disease Control and Prevention (U.S.); Characteristics; Chronic; Chronic Childhood Arthritis; Clinical; Collagen; Collagen Type II; Communities; Cyclophosphamide; DNA; DNA Binding; DNA Sequence; Data; Degenerative polyarthritis; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Disease Progression; Early Diagnosis; Economic Burden; Effectiveness of Interventions; Epitopes; Exhibits; Extracellular Matrix; Funding; Goals; Gout; Health Professional; Individual; Inflammation; Joints; Label; Libraries; Life Style; Ligands; Mammals; Measures; Medical; Methods; Molecular Structure; Monitor; Monoclonal Antibodies; National Institute of Arthritis and Musculoskeletal and Skin Diseases; Patients; Peptides; Persons; Phase; Process; Production; Prognostic Marker; Quality of life; Relative (related person); Reporting; Rheumatoid Arthritis; Serum; Signal Transduction; Silicon; Staging; Structure; Symptoms; Testing; Therapeutic; Time; Treatment Efficacy; United States; Urine; Validation; Work; base; chemokine; clinically relevant; cost; cytokine; design; design and construction; disability; innovation; joint destruction; nanowire; novel; outcome forecast; palliative; prognostic; public health relevance; research study; urinary; user-friendly

DESCRIPTION (provided by applicant): Arthritis (AR) is the leading cause of disability in the United States, with 50 million Americans (22%) suffering from some form of the disease or chronic joint symptoms. The CDC estimates that, by the year 2030, 67 million adults will have AR and the economic burden will reach an annual cost of $128 billion dollars. While most AR cases are classified as osteoarthritis (OA), affecting over 27 million Americans, other forms of the disease are also prevalent, including rheumatoid arthritis, juvenile arthritis and gout. AR is associated with the breakdown of collagen and the production of inflammation-related factors such as cytokines. The degradation of Type I and Type II collagens in AR patients results in the presence of detectable peptide biomarkers in urine; these include CTX-II, CTX-I and C2C. The levels of these biomarkers in urine track with the extent of cartilage damage in patients. The Osteoarthritis Biomarkers Network has classified these biomarkers regarding their applicability to OA diagnosis, prognosis and efficacy of treatment. Because AR can progress slowly, and often patients with cartilage degeneration are asymptomatic for years, diagnosing AR in its early stages and monitoring its progression has been a major challenge. We propose to develop a non-invasive, highly sensitive diagnostic assay for AR using DNA origami molecules that can detect these three biomarkers. In Phase I, we will evaluate the feasibility of performing DNA origami selections against the CTX-II, CTX-I and C2C peptides to identify DNA sequences that we will develop as probes in an assay to specifically diagnose AR. Three aims are proposed: AIM1: Design and construct a DNA origami library for AR biomarker selections. AIM 2: Isolate DNA origami molecules that interact with AR biomarker peptides. AIM 3: Identify and characterize DNA origami molecules that bind the AR biomarkers. Due to the DNA origami molecular structure, the interaction between these molecules and the AR biomarkers should exhibit increased binding affinities, which contributes significantly to the sensitivity and ingenuty of this diagnostic assay. This assay will provide the medical community with a reliable, non-invasive test for AR that offers increased sensitivity and convenience relative to current assays. Following successful completion of our Phase I aims, this work will continue into Phase II, with the development of a label-free, highly sensitive detection assay, using silicon nanowires to signal the presence of these biomarkers in urine. The development of a non-invasive multiplex diagnostic assay for AR has the potential to result in a clinical paradigm shift from therapies tha are largely palliative to one that focuses on identifying persons with early stage AR. The assay will also have a prognostic and treatment efficacy value for monitoring cartilage breakdown in patients receiving various treatments. Further, this assay will provide the necessary surveillance and an excellent vehicle to motivate afflicted individuals to make lifestyle-changing decisions (e.g. weight loss) to increase their quality of life. Lastly, if the feasibility of the method is demonstrated, the method could be applied to other candidate biomarkers in AR.

描述（由申请人提供）：关节炎 (AR) 是美国残疾的主要原因，有 5000 万美国人 (22%) 患有某种形式的疾病或慢性关节症状。 CDC估计，到2030年，将有6700万成年人患有AR，每年的经济负担将达到1280亿美元。虽然大多数 AR 病例被归类为骨关节炎 (OA)，影响超过 2700 万美国人，但其他形式的疾病也很普遍，包括类风湿性关节炎、幼年关节炎和痛风。 AR 与胶原蛋白的分解和细胞因子等炎症相关因子的产生有关。 AR 患者 I 型和 II 型胶原蛋白的降解导致尿液中出现可检测的肽生物标志物；其中包括 CTX-II、CTX-I 和 C2C。尿液中这些生物标志物的水平与患者软骨损伤的程度密切相关。骨关节炎生物标志物网络根据这些生物标志物对 OA 诊断、预后和治疗效果的适用性对其进行了分类。由于 AR 进展缓慢，而且软骨退变患者通常多年没有症状，因此在早期阶段诊断 AR 并监测其进展一直是一项重大挑战。我们建议使用 DNA 折纸分子开发一种非侵入性、高灵敏度的 AR 诊断方法，可以检测这三种生物标志物。在第一阶段，我们将评估针对 CTX-II、CTX-I 和 C2C 肽进行 DNA 折纸选择的可行性，以确定我们将开发的 DNA 序列，作为特异性诊断 AR 的检测中的探针。提出了三个目标： AIM1：设计和构建用于 AR 生物标志物选择的 DNA 折纸库。目标 2：分离与 AR 生物标记肽相互作用的 DNA 折纸分子。目标 3：识别并表征结合 AR 生物标志物的 DNA 折纸分子。由于 DNA 折纸分子结构，这些分子和 AR 生物标志物之间的相互作用应该表现出增加的结合亲和力，这对这种诊断测定的灵敏度和独创性有很大贡献。该检测将为医学界提供一种可靠的、非侵入性的 AR 测试，相对于当前的检测，它具有更高的灵敏度和便利性。在成功完成第一阶段目标后，这项工作将继续进入第二阶段，开发一种无标记、高灵敏度的检测方法，使用硅纳米线来发出尿液中这些生物标志物存在的信号。针对 AR 的非侵入性多重诊断检测的开发有可能导致临床范式的转变，从很大程度上是姑息治疗到专注于识别早期 AR 患者的治疗。该测定还将具有预后和治疗功效价值，用于监测接受各种治疗的患者的软骨破坏。此外，该测定将提供必要的监测和良好的工具，以激励受影响的个体做出改变生活方式的决定（例如减肥）以提高他们的生活质量。最后，如果该方法的可行性得到证实，该方法可以应用于 AR 中的其他候选生物标志物。