Cell Model & Assay Core

细胞模型

基本信息

批准号：
8451288
负责人：
KEVIN L KIRK
金额：
$ 22万
依托单位：
UNIVERSITY OF ALABAMA AT BIRMINGHAM
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-05-01 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8451288
关键词：
Area Binding Biochemical Biological Assay Cell Culture Techniques Cell Line Cell model Cells Consultations Core Facility Cost Savings Country Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Development Disease Epithelial Cells Equipment Fostering Funding Hereditary Disease Human Resources Individual Interdisciplinary Study Investigational Therapies Laboratories Laboratory Research Lung Morbidity - disease rate Mus Mutation Patch-Clamp Techniques Pathogenesis Peptides Performance Pharmaceutical Preparations Post-Translational Protein Processing Proteins Proteomics Protocols documentation Reading Reagent Regulation Research Research Personnel Resources Subfamily lentivirinae Techniques Testing Training Translational Research United States National Institutes of Health Validation base cellular transduction cystic fibrosis airway effective therapy efficacy testing high throughput screening improved innovation monolayer mortality mutant next generation novel patch clamp prospective ranpirnase research study

Area Binding Biochemical Biological Assay Cell Culture Techniques Cell Line Cell model Cells Consultations Core Facility Cost Savings Country Cystic Fibrosis Cystic Fibrosis Transmembrane Conductance Regulator Development Disease Epithelial Cells Equipment Fostering Funding Hereditary Disease Human Resources Individual Interdisciplinary Study Investigational Therapies Laboratories Laboratory Research Lung Morbidity - disease rate Mus Mutation Patch-Clamp Techniques Pathogenesis Peptides Performance Pharmaceutical Preparations Post-Translational Protein Processing Proteins Proteomics Protocols documentation Reading Reagent Regulation Research Research Personnel Resources Subfamily lentivirinae Techniques Testing Training Translational Research United States National Institutes of Health Validation base cellular transduction cystic fibrosis airway effective therapy efficacy testing high throughput screening improved innovation monolayer mortality mutant next generation novel patch clamp prospective ranpirnase research study

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项目摘要

Cystic fibrosis investigators within the P30 Center require access to expertise and specialized equipment for their studies of CFTR expression and function. For both established CF laboratories and those new to CF research, a need has been identified for developing novel cell lines expressing mutant and wild type CFTR, and for dedicated equipment and biophysical protocols necessary to assess CFTR function. In particular, we propose two specific aims: Specific Aim 1. Cellular models expressing wild type and mutant CFTR will be developed and provided by Core A. These will include primary cells from murine lungs and novel lentiviral-transduced cell lines. The latter include innovative cell models useful for CFTR proteomic and structural studies and for high throughput screens to discover the next generation of CF drugs. Specific Aim 2, Assistance, equipment and expertise necessary to perform functional assays of CFTR in the above cell models (Aim 1) will include biophysical techniques (patch clamp). These assays are required to test the efficacies of new maneuvers to rescue mutant CFTR channel activity and regulation as well as to define the underlying mechanisms. The Core will aid in the development/validation of new CF cell models; provide primary murine airway epithelial cells encoding specific CFTR mutations; and assist investigators with their experiments to test the effects of new maneuvers on mutant CFTR (e.g., DeltaF508) protein stability and channel function. Collaborative studies involving proteomic studies of CFTR post-translational modifications, discovery of peptides from the first cytosolic loop of CFTR that specifically block NBD1-TMD1 binding and numerous other NIH funded projects will also be assisted by the Core. Core A will foster interdisciplinary research by providing valuable new cell models and assays of CFTR expression and function to investigators less familiar with the requisite techniques, and contribute to innovative studies of CF pathogenesis and experimental therapy.

P30中心内的囊性纤维化研究者需要使用专业知识和专业设备来研究CFTR的表达和功能。对于已建立的CF实验室和新手CF研究的实验室，已经确定了开发表达突变体和野生型CFTR的新型细胞系，以及评估CFTR功能所需的专用设备和生物物理方案。特别是，我们提出了两个具体目标：具体目的1。表达野生型和突变体CFTR的细胞模型将由核心A开发并提供。这些将包括鼠肺和新型慢病毒转导的细胞系的原代细胞。后者包括可用于CFTR蛋白质组学和结构研究的创新细胞模型以及高吞吐量筛选以发现下一代CF药物。在上述细胞模型中执行CFTR功能测定所需的特定目标2，辅助，设备和专业知识（AIM 1）将包括生物物理技术（贴片夹）。这些测定需要测试新操纵的效力以挽救突变体CFTR通道活性和调控以及定义潜在机制。核心将有助于开发/验证新的CF细胞模型；提供编码特定CFTR突变的主要鼠气上皮细胞；并协助研究人员进行实验，以测试新操纵对突变CFTR（例如Deltaf508）蛋白质稳定性和通道功能的影响。涉及CFTR翻译后修饰的蛋白质组学研究的协作研究，从CFTR的第一个胞质环中发现肽，这些肽特异性阻断NBD1-TMD1结合以及许多其他NIH资助的项目也将得到核心的帮助。核心A将通过为不熟悉必要技术的研究人员提供宝贵的新细胞模型和CFTR表达和功能的测定来促进跨学科研究，并为CF发病机理和实验疗法的创新研究做出贡献。