SBIR Phase I: Development and Validation of an In Vitro Diagnostic of HDL Cholesterol Efflux and Coronary Artery Disease Risk

这个小型企业创新研究(SBIR)第一阶段项目将开发一种准确预测冠状动脉疾病(CAD)风险的新方法。由于高密度脂蛋白(HDLE)可预防冠心病，目前的风险评估方法是测量HDL胆固醇含量(HDLC)，以此作为HDL量的替代指标。然而，高密度脂蛋白的功能，即其排出胆固醇的能力，是降低冠心病风险的原因。肥胖和糖尿病易患冠心病，并影响高密度脂蛋白的外流活性。由于直接定量测定胆固醇流出活性不符合临床实验室的要求，我们开发了一种利用电子顺磁共振波谱间接报告高密度脂蛋白-S胆固醇流出活性的新方法。我们的分析方法对小于10微米的L血浆/血清在大约10分钟内完成，适合常规自动化、高通量临床使用。目标是：1)与直接测量高密度脂蛋白胆固醇外流活性相比较，验证EPR检测方法；2)确定我们的检测方法在已建立的2型糖尿病(METSIM)患者队列中识别功能障碍的高密度脂蛋白的预测能力。预计该分析方法在确定CAD风险方面将具有优于所有现有方法的灵敏度和预测能力。该项目更广泛的影响/商业潜力是实现对高密度脂蛋白功能的快速评估并准确预测CAD风险，这在医疗诊断和药物开发领域是一个数十亿美元的商业机会。该项目将开发和验证一种报告高密度脂蛋白功能的快速、灵敏和准确的检测方法。这种创新的(一流的)、不连续的(超越高密度脂蛋白-C)和转化试验(直接在血浆中对高密度脂蛋白功能的高度特异性测量)可以被用作一种诊断分析来识别处于危险中的患者，即使是那些高密度脂蛋白水平正常的患者，也可以作为临床试验和患者管理中的替代结果衡量标准。我们的检测以一种与年龄和性别无关的方式作为代谢性疾病状态的窗口，其非凡的能力是前所未有的。这项技术有望极大地提高对冠心病和代谢性疾病易感性的早期诊断，促进发现和实施预防疾病的干预措施，并显著降低长期保健成本。EPR光谱作为疾病的功能生物标志物更广泛地用于测量蛋白质的结构变化，这是一项平台创新，标志着分子诊断学进入了一个复杂的新时代。

This Small Business Innovation Research (SBIR) Phase I project will develop a new approach to accurately predict coronary artery disease (CAD) risk. Because high-density-lipoprotein (HDL) protects from CAD, current methods to assess risk measure the cholesterol content of HDL (HDL-C) as a surrogate indicator of HDL quantity. However, the function of HDL, namely its ability to efflux cholesterol, is what reduces CAD risk. Obesity and diabetes predispose to CAD and compromise HDL cholesterol efflux activity. Because direct quantification of cholesterol efflux activity is not amenable to clinical laboratories, we developed a novel assay that uses electron paramagnetic resonance (EPR) spectroscopy to report indirectly on HDL?s cholesterol efflux activity. Our assay is performed in approximately 10 minutes on less than 10 µl of blood plasma/serum and is suitable for routine automated, high throughput clinical use. The objectives are: 1) validate the EPR assay compared to direct measurement of HDL cholesterol efflux activity, and 2) determine the predictive power of our assay to identify dysfunctional HDL in an established cohort of individuals with type 2 diabetes (METSIM). It is expected that this assay will have superior sensitivity and predictive power over all existing methods to determine CAD risk.The broader impact/commercial potential of this project is to enable rapid assessment of HDL function and accurately predict CAD risk, a multi-billion-dollar commercial opportunity in the medical diagnostics and drug development space. This project will develop and validate a rapid, sensitive and accurate assay that reports on HDL function. This innovative (first-in-class), discontinuous (goes beyond HDL-C), and transformational assay (highly specific measurement of HDL function directly in plasma) can be used as a diagnostic assay to identify patients at risk, even those with normal HDL-C levels, and as a surrogate outcome measure in clinical trials and patient management. The exceptional capability of our assay to serve as a window on metabolic disease status in an age- and gender-independent fashion is unprecedented. This technology promises to greatly improve the early diagnosis of CAD and metabolic disease predisposition and facilitate the discovery and implementation of interventions to prevent disease and significantly reduce long-term healthcare costs. The still broader use of EPR spectroscopy to measure structural changes in proteins as functional biomarkers of disease represents a platform innovation that denotes a new era of sophistication for molecular diagnostics.