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：
8822868
负责人：
Chang Lu
金额：
$ 32.46万
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/8822868
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 Health 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 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 research study single molecule temporal measurement 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）：诸如小鼠之类的实验室动物是了解疾病发展，测试候选药物和设计治疗的关键工具。尽管产生了大量知识，但对小鼠的分子生物学测定主要是通过集合测量组的平均特性进行的。但是，与单分子研究背后的基本原理类似，整体平均值通常掩埋有关动态的重要细节，而忽略了种群异质性和亚集。在该项目中，我们将基于“单个活动物实验”的疾病发育期间研究基因法规的时间动力学（即内毒素引起的动脉粥样硬化）。我们将根据小鼠的少量血液样本进行测试，开发超敏感的微流体CHIP-QPCR和芯片SEQ分析。原则上，这些测试可能是最小的侵入性，并且不会扰动动物的状态和疾病过程。在疾病发展过程中，我们将能够定期检查同一活小鼠，并了解转录因子/启动子结合和组蛋白修饰中的时间动力学。我们认为，单一活跃数据将对这些生物学过程中涉及的分子事件的独特见解，并为诊断，预后，药物设计/发现和治疗策略提供重要的基础。这些数据也最紧密地模仿了疾病发育和治疗期间人类患者发生的情况，因此具有直接的临床相关性。