Microfluidic assays for hyper-reactive platelets in diabetes

糖尿病高反应性血小板的微流控检测

• 批准号: 9199213
• 负责人: Hang Lu
• 金额: $ 22.8万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9199213
• 关键词: Adhesions; Adopted; Affinity; Agonist; Automation; Behavior; Binding; Biochemical; Biochemical Process; Biological Assay; Biology; Biomechanics; Blood; Blood Platelet Disorders; Blood Platelets; Blood specimen; Cardiovascular Diseases; Cell Adhesion; Cigarette Smoker; Clinical; Collaborations; Complex; Coupled; Data; Diabetes Mellitus; Diagnosis; Diagnostic; Disease; Disease Progression; Drops; Engineering; Event; Future; Goals; Hyperactive behavior; Hypertension; Integrins; Kinetics; Knowledge; Lead; Literature; Microfluidics; Modification; Molecular; Molecular Conformation; Molecular Probes; Morphology; Obesity; P-Selectin; Pathologic; Patients; Pattern; Physicians; Platelet Activation; Platelet Count measurement; Play; Process; Regulation; Reporter; Resolution; Risk Factors; Role; Shapes; Surface; System; Technology; Testing; Thrombosis; Time; Up-Regulation; Validation; Work; atherothrombosis; base; clinical Diagnosis; clinical application; design; diabetic; diabetic patient; experience; experimental study; high throughput screening; image processing; innovation; insight; public health relevance; receptor; tool; von Willebrand Factor

 DESCRIPTION (provided by applicant): Diabetes, obesity, and hypertension among other cardiovascular diseases are large risk factors for platelet hyperreactivity. Despite the importance, the molecular mechanisms of the platelet hyperreactivity are still unknown. Our long-term goal is to integrate biomechanical and biochemical approaches to understand the disease mechanisms in patients with diabetes, obesity and cardiovascular diseases, and to use this knowledge to design tools that facilitate physicians' decisions on treatment of these diseases - tools to diagnose, tools to follow disease progression, and tools to follow treatment courses. Using a unique single-platelet Biomembrane Force Probe (BFP) assay, we have gathered preliminary evidence that there exists an intermediate state of platelets (discoid in shape but express low-level markers of activation) and this state is primarily characterized by having integrin molecules adopting a conformation that gives rise to intermediate affinity. We hypothesize that this intermediate state plays an important role in platelet hyperactivity in diabetics. While this assay is sensitive and powerful for probing molecular interactions on single platelets, it is very labor-intensive and low throughput. The goal of this project is to design a simple-to-use and yet high-throughput and highly informative microfluidic approach to understand sequences of molecular events that lead to platelet activation. We will obtain detailed characterization of the intermediate state, its stability, and the kinetics of state changs of the normal and diseased platelets using this approach. Validation of the new assay and proof of the hyperactivity hypothesis will allow this assay to be further developed in the future for clinical diagnosis or to follow treatment of atherothrombosis in patients. We have assembled a team of engineers and clinicians for this project. The work is innovative because no such high-throughput assay that yields mechanistic insights (and only using a drop of blood) is currently available, and that understanding the role of the intermediate state of platelets, particularly in diabetes, will lead to a significant improvement in diagnostic and treatment for platelet hyperactivity disorders.

 描述（由申请人提供）：糖尿病、肥胖症和高血压等心血管疾病是血小板高反应性的重要危险因素。尽管很重要，但血小板高反应性的分子机制仍然未知。我们的长期目标是整合生物力学和生化方法来了解糖尿病、肥胖症和心血管疾病患者的疾病机制，并利用这些知识来设计有助于医生做出治疗这些疾病决策的工具——诊断工具、跟踪疾病进展的工具以及跟踪治疗过程的工具。使用独特的单血小板生物膜力探针（BFP）测定，我们收集了初步证据，表明存在血小板的中间状态（盘状但表达低水平的激活标记物），并且这种状态的主要特征是整合素分子采用产生中间亲和力的构象。我们假设这种中间状态在糖尿病患者血小板过度活跃中发挥重要作用。虽然这种测定对于探测单个血小板上的分子相互作用非常敏感且强大，但它非常劳动密集型且通量低。该项目的目标是设计一种易于使用、高通量且信息丰富的微流体方法，以了解导致血小板激活的分子事件序列。我们将使用这种方法获得正常和患病血小板的中间状态、其稳定性以及状态变化动力学的详细表征。新测定的验证和多动假说的证明将使该测定在未来得到进一步开发，用于临床诊断或跟踪患者动脉粥样硬化血栓形成的治疗。我们为这个项目组建了一个由工程师和临床医生组成的团队。这项工作具有创新性，因为目前还没有这样的高通量测定法可以产生机制见解（并且仅使用一滴血），并且了解血小板中间状态的作用，特别是在糖尿病中，将导致血小板过度活跃性疾病的诊断和治疗的显着改善。