Probing dynamics in protein-DNA interactions during disease development using sin

描述(由申请人提供)：小鼠等实验室动物是了解疾病发展、测试候选药物和设计治疗方法的关键工具。尽管产生了大量的知识，但对小鼠的分子生物学分析大多是通过对一组人的平均性质进行整体测量来完成的。然而，类似于单分子研究背后的理论基础，总体平均往往掩盖了关于动力学的重要细节，而忽略了群体异质性和子集。在本项目中，我们将在“单一活体动物实验”的基础上，研究疾病发展(即内毒素血症诱导的动脉粥样硬化)过程中基因调控的时间动力学。我们将开发超灵敏的微流控芯片-qPCR和芯片-序列分析，用于基于微量小鼠血液样本的检测。原则上，这些测试可以是微创的，不会扰乱动物的状态和疾病过程。我们将能够在疾病发展过程中对同一只活鼠进行定期检查，并了解转录因子/启动子结合和组蛋白修饰的时间动力学。我们相信，单个活体动物数据将为这些生物过程中涉及的分子事件提供独特的见解，并为诊断、预后、药物设计/发现和治疗策略提供重要依据。这些数据也最接近地模拟了人类患者在疾病发展和治疗过程中发生的情况，从而提供了直接的临床相关性。

DESCRIPTION (provided by applicant): Lab animals such as mice are critical tools for understanding the disease development, testing drug candidates, and devising treatments. In spite of the vast amount of knowledge generated, molecular biology assays on mice have mostly been done via ensemble measurements of the average properties of a group. However, similar to the rationale behind single molecule studies, ensemble averages often bury important details about the dynamics and ignore population heterogeneity and subsets. In this project, we will study the temporal dynamics in gene regulations during disease development (i.e. endotoxemia-induced atherosclerosis) based on "single live animal experiments". We will develop ultrasensitive microfluidic ChIP-qPCR and ChIP- seq assays for testing based on tiny amounts of blood samples from mice. In principle these tests can be minimally invasive and do not perturb the state of the animal and the disease process. We will be able to conduct periodical examination of the same live mouse over the course of the disease development and understand the temporal dynamics in the transcription factor/promoter bindings and histone modifications. We believe that the single live animal data will grant unique insights into the molecular events involved in these biological processes and provide important basis for diagnosis, prognosis, drug design/discovery, and treatment strategy. Such data also most closely mimic what occurs in human patients during disease development and treatment, thus offer direct clinical relevance.