Role of Single Cell mRNA Variation in Systems Associated Electrically Excitable C

单细胞 mRNA 变异在与电兴奋 C 相关的系统中的作用

基本信息

批准号：
8549306
负责人：
JAMES H EBERWINE
金额：
$ 187.76万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-19 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8549306
关键词：
Accounting Action Potentials Agonist Algorithms Arrhythmia Biological Biological Process Biophotonics Cardiac Cardiac Myocytes Cell physiology Cells Central Nervous System Diseases Cessation of life Collaborations Complementary DNA Contracts Data Data Set Dimensions Disease Environment Functional disorder G Protein-Coupled Receptor Genes G-Protein-Coupled Receptors Gene Expression Gene Expression Profile Genes Goals Heart Heterogeneity Human Human body In Situ Individual Introns Lead Life Maps Measures Mediating Messenger RNA Methodology Molecular Muscle Cells Neurons Neurosurgeon Noise Pathway interactions Phenotype Physiological Physiology Procedures RNA RNA Splicing Role Scientist Slice Source Specimen System Techniques Technology Testing Therapeutic Tissue Sample Tissues Transcript Untranslated Regions Variant base biobank cell type combinatorial design functional genomics in vivo novel public health relevance response tissue culture

项目摘要

DESCRIPTION (provided by applicant): The goal of this U01 is to characterize and understand the variability in the expressed transcriptome of human excitable cells. There are two predominant types of excitable cell in the human body, neurons and muscle cells, including cardiac cells. Many human CNS diseases result from modulation of the electrical responsiveness of neurons while cardiac arrhythmias account for most of heart associated deaths. However, at the level of individual cells there is considerable heterogeneity in function, response, and dysfunction. Here, we present preliminary data showing large-scale single cell variability that is difficult to explain as simple molecular noise. We hypothesize that there is a many-to-one relationship between transcriptome states and a cell's phenotype. In this relationship the functional molecular ratios of the RNA are determined by the cell systems' stoichiometric constraints, which underdetermine the transcriptome state. Because a broad set of multi-genic combinations support a particular phenotype, changes in the transcriptome state do not necessarily lead to changes in the phenotype potentially explaining cellular heterogeneity in phenotype response to variant conditions such as the application of therapeutic molecules. To test this hypothesis we propose to investigate the extent of single cell variation for the whole transcriptome for excitable cells that are in their natural environment using a novel mRNA capture methodology (TIVA-tag), and on a subset of the transcriptome, the mRNAs encoding the therapeutically important and manipulable G protein-coupled receptor (GPRC) pathways. The use of functional genomics techniques developed in the Eberwine and Kim labs (TIPeR) will permit an assessment of the biological role of multigenic transcriptome variation. These studies are truly interdisciplinary involving the collaboration of two clinicians (Drs. Grady, Neurosurgeon and Kuhn, Cardiologist), two genomicists (Drs. Eberwine and Kim) one of whom is a computational scientist (Dr. Kim), a neuro/cardio- pharmacologist (Dr. Bartfai) and a biophotonics expert (Dr. Sul).!

描述（由申请人提供）：该U01的目的是表征和了解人类兴奋性细胞表达的转录组的变异性。人体，神经元和肌肉细胞（包括心脏细胞）中有两种主要类型的可兴奋细胞类型。许多人类中枢神经系统疾病是由神经元的电反应能力的调节引起的，而心律不齐是大多数与心脏相关的死亡。但是，在单个细胞的水平上，功能，响应和功能障碍存在相当大的异质性。在这里，我们提供了显示大规模单细胞变异性的初步数据，这很难解释为简单的分子噪声。我们假设转录组状态与细胞表型之间存在许多一对一的关系。在这种关系中，RNA的功能分子比由细胞系统的化学计量限制确定，这些约束不足。由于一组广泛的多类别组合支持特定的表型，因此转录组状态的变化不一定会导致表型的变化，这可能会在表型反应中可能解释了对变异条件（例如应用治疗分子的应用）中细胞异质性的变化。为了检验该假设，我们建议研究整个单细胞变异的程度使用新型的mRNA捕获方法（TIVA-TAG）以及转录组的子集，编码治疗上重要且可操作的G蛋白偶联受体（GPRC）途径的mRNA，使用新型mRNA捕获方法（TIVA-TAG）以及在自然环境中的转录组。在Eberwine和Kim Labs（TIPER）中开发的功能基因组学技术的使用将允许评估多基因转录组变化的生物学作用。这些研究确实是跨学科的，涉及两位临床医生的合作（Grady博士，神经外科医生心脏病专家库恩（Kuhn），两位基因组学家（Eberwine和Kim博士），其中一位是一名计算科学家（Kim博士），神经/心脏药理学家（Bartfai博士）和生物原理专家（SUL博士）。