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

• 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
• 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

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）：实验室动物（如小鼠）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测定大多是通过对一组平均特性的整体测量来完成的。然而，类似于单分子研究背后的基本原理，系综平均值往往掩盖了有关动力学的重要细节，忽略了群体异质性和子集。在这个项目中，我们将研究在疾病发展过程中的基因调控的时间动态（即内毒素血症诱导的动脉粥样硬化）基于“单活动物实验”。我们将开发超灵敏的微流控ChIP-qPCR和ChIP-seq检测方法，用于基于小鼠的微量血液样本进行测试。原则上，这些测试可以是微创的，并且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学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期间发生的情况，因此提供了直接的临床相关性。