Microfluidic tools for probing neutrophil reversed migration

描述(申请人提供)：发达国家十大主要死亡原因中有七个(从动脉粥样硬化和心脏病到糖尿病和癌症)都有一个共同的病理：慢性炎症。诊断通常是当关键器官的损害已经是显著的和不可逆转的，因为现有的炎症标志物都不适合早期检测慢性炎症状况。为了改变现状，我们将依靠新兴知识提出中性粒细胞在慢性炎症中的新角色--中性粒细胞是保护我们免受微生物侵袭的典型先天免疫细胞。在转基因动物中的观察表明，一些中性粒细胞能够在对无菌损伤做出反应后逆转从组织到循环的迁移。最近，我们已经能够在一种新型的体外装置上复制反向迁移模式。这项提议的主要目标是继续改进这些设备的设计，识别和表征体外反向迁移的调节器，并设计更复杂的技术来分离经历反向迁移的中性粒细胞，用于随后的基因组和蛋白质组分析。这些工具将使我们能够详细描述反向迁移的中性粒细胞的特征，并识别特定的标记物，可以区分慢性炎症患者血液样本中的反向迁移的中性粒细胞和幼稚的中性粒细胞。如果成功，我们的研究可能会带来监测慢性炎症的新方法，可以进行早期诊断，留出足够的时间改变生活方式，协助早期治疗，并可能对降低与慢性炎症疾病相关的发病率和死亡率产生重大影响。

DESCRIPTION (provided by applicant): Seven out of ten top leading causes of mortality in the developed world (from atherosclerosis and heart disease to diabetes and cancer) share a common pathology: chronic inflammation. Diagnosis is often made when the damage to critical organs is already significant and irreversible because none of the existing inflammation markers is appropriate for early detection of chronic inflammation conditions. To change the status-quo, we will rely on emerging knowledge proposing new roles for neutrophils - the typical innate immune cell to protect us against microbes - in chronic inflammation. Observations in transgenic animals showed that some neutrophils are capable of reversing migration from tissues to circulation after responding to a sterile injury. Recently, we have been able to replicate the reversed-migration patterns in a novel in vitro device. The major goal of this proposal is to continue improving the design of these devices, to identify and characterize the modulators of reversed-migration in vitro, and to design more sophisticated technologies that can separate the neutrophils undergoing reversed migration, for subsequent genomic and proteomic analysis. These tools would enable us to characterize the reversely-migrated neutrophils in detail and to identify specific markers that could differentiate the reversely-migrated neutrophils from naive neutrophils in blood samples from patients with chronic inflammation. If successful, our research could lead to new methods for monitoring chronic inflammation, could enable early diagnosis, allow sufficient time to adopt lifestyle changes, assist early treatments, and could have major implications for reducing morbidity and mortality associated with chronic inflammation diseases.