Rapid Microfluidic-Based Point-of-Care Assessment of Epigenetic Marks for Diagnosis, Prognosis and Response to Therapy in Critical Illness

败血症是由对感染的压倒性和不平衡反应引发的全身炎症和器官衰竭，会造成巨大的人力和医疗成本。在全球范围内，每年有3000多万人受到影响，其中至少四分之一将死亡。这些后遗症包括身体上、精神上和心理上的残疾。目前还没有具体的治疗方法。对于癌症患者来说，基于肿瘤分子特征的个性化治疗是一种很好的做法。对于脓毒症，迫切需要采取同样的措施。我们已经确定了DNA上的标签（DNA甲基化标记），可以区分脓毒症和非脓毒症患者，不良结果（死亡和器官衰竭）的可能性以及对治疗（血压药物）的反应。与此同时，我们的合作伙伴已经开发出一种使用创新的微流体方法从全血中分离DNA的即时护理设备并获得专利。我们建议采用这项技术开发一种原型，能够使用全血对患者的DNA标记进行自动测量；用DNA和全血样本测试我们的设备；使用预测模型整合临床和分子数据，以验证独立队列的结果预测和治疗反应；并对原型的性能和精度进行了实际测试。这项技术和生物标志物检测方法之前从未被用于败血症。这项提案汇集了生物工程师、健康研究科学家、生物信息学家、医生和临床生物化学家。成功完成将提供一种采用低成本工业制造的聚合物微流控设备的芯片实验室技术，该技术由先进的专利离心微流控技术驱动，有望成为整合和部署基因组知识实用设备的颠覆性创新，将影响临床实践和危重病人的护理。

Sepsis, whole body inflammation and organ failure triggered by an overwhelming andunbalanced response to infection, incurs enormous human and health care costs. Globally,over 30 million people are affected each year, at least a quarter of whom will die. Thesequelae include physical, mental, and psychological disability. Currently, no specifictreatments exist. For cancer patients, personalizing treatment on the basis of molecularcharacteristics of a tumor is simply good practice. There is an urgent need to do the same forsepsis. We have identified tags on DNA (DNA methylation marks) that can classify septic vs.non-septic patients, the likelihood of poor outcomes (death and organ failure) and response totherapy (blood pressure medications). In parallel, our partners have developed and patenteda point-of-care device that uses an innovative microfluidic approach to isolate DNA fromwhole blood. We propose to adapt this technology to develop a prototype able to performautomated measurements of DNA marks from patients using whole blood; to test our deviceon banked DNA and whole blood samples; to use predictive modeling to integrate clinical andmolecular data to validate outcome prediction and response to therapy in an independentcohort; and to test the performance of our prototype and precision marks prospectively in areal-world situation. Both the technology and biomarker detection approaches have neverbeen tried for sepsis before. This proposal brings together bioengineers, health researchscientists, bioinformaticians, physicians and clinical biochemists. Successful completion willprovide a Lab-on-chip technology employing low-cost industrially manufacturable polymermicrofluidic devices powered by an advanced patented centrifugal microfluidic technology thatpromises to be a disruptive innovation in the way to integrate and deploy genomic knowledgepractical devices that will impact clinical practices and the care of the critically ill.