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

使用 sin 探索疾病发展过程中蛋白质-DNA 相互作用的动态

基本信息

批准号：
8697316
负责人：
Chang Lu
金额：
$ 34.65万
依托单位：
VIRGINIA POLYTECHNIC INST AND ST UNIV
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8697316
关键词：
Animal Diseases Animal Experiments Animals Atherosclerosis Binding Biological Assay Biological Process Blood Blood specimen Cell Line Cells DNA DNA-Protein Interaction Data Development Devices Diagnosis Disease Disease Progression Drug Design Endotoxemia Epigenetic Process Event Gene Expression Gene Expression Regulation Genes Genome Grant Histones Human In Vitro Inbred Mouse Individual Knowledge Life Maps Measurement Measures Microfluidic Microchips Microfluidics Molecular Molecular Biology Mus Pathogenesis Patients Pattern Periodicals Population Heterogeneity Process Property Protocols documentation Resolution Sample Size Sampling Site Technology Testing Time Variant animal data base chromatin immunoprecipitation clinically relevant drug candidate genome-wide genome-wide analysis histone modification insight interest minimally invasive monocyte outcome forecast promoter public health relevance research study single molecule therapy development tool transcription factor treatment strategy

项目摘要

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.

描述（由申请人提供）：小鼠等实验动物是了解疾病发展、测试候选药物和设计治疗方法的重要工具。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平均特性的整体测量来完成的。然而，与单分子研究背后的基本原理类似，整体平均值常常隐藏有关动态的重要细节，并忽略群体异质性和子集。在这个项目中，我们将基于“单一活体动物实验”来研究疾病发展（即内毒素血症引起的动脉粥样硬化）期间基因调控的时间动态。我们将开发超灵敏的微流控 ChIP-qPCR 和 ChIP-seq 检测方法，用于基于小鼠微量血液样本进行测试。原则上，这些测试可以是微创的，不会干扰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小鼠进行定期检查，并了解转录因子/启动子结合和组蛋白修饰的时间动态。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，因此提供了直接的临床相关性。