Na+ TRANSPORT INHIBITION BY RESPIRATORY SYNCYTIAL VIRUS

呼吸道合胞病毒对 Na 转运的抑制

• 批准号: 7110384
• 负责人: IAN CHRISTOPHER DAVIS
• 金额: $ 12.01万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7110384
• 关键词: A549; Alveolar; Amiloride; Anatomy; Animal Model; Animals; Apoptosis; Biochemical; Bronchiolitis; Cell physiology; Cell surface; Cells; Child; Clinical; Cytopathology; Development; Doctor of Philosophy; Electrophysiology (science); Ensure; Environment; Epithelial; Epithelial Cells; Human; Hypoxemia; In Situ Hybridization; In Vitro; Inbred BALB C Mice; Infection; Ion Channel; Ion Transport; Ions; Knowledge; Laboratory Animal Medicine; Laboratory Animals; Liquid substance; Lower respiratory tract structure; Lung; Marines; Measurement; Measures; Mediating; Mentors; Mining; Mus; Necrosis; Northern Blotting; Nose; Patch-Clamp Techniques; Pathogenesis; Pathology; Pathway interactions; Physiology; Property; Protein Subunits; Proteins; Research; Respiratory Syncytial Virus Infections; Respiratory Tract Diseases; Respiratory physiology; Respiratory syncytial virus; Reverse Transcriptase Polymerase Chain Reaction; Ribavirin; Role; Scientist; Seminal; Structure of respiratory epithelium; Techniques; Testing; Trachea; Training; Ubiquitin; Universities; Upper respiratory tract; Viral; Viral Pathogenesis; Virus; Western Blotting; alveolar epithelium; comparative; epithelial Na+ channel; in vitro Model; in vivo; monolayer; multicatalytic endopeptidase complex; patch clamp; protein expression; respiratory; respiratory virus; stoichiometry; veterinary science

CANDIDATE: I graduated from the University of Bristol in 1992, with Honours in Anatomical Science and Veterinary Science. At UAB I received training in laboratory animal medicine, laboratory animal pathology, and viral pathogenesis in animal models (Ph.D. February 2000; mentor Dr. Patricia Fultz). Subsequently, I elected to join Dr. Matalon's research group, where I might build on my knowledge of viral pathogenesis by cross-mining in electrophysiology, and apply these techniques to study the pathogenesis of human respiratory viral pneumonitides. RESEARCH: Respiratory syncytial virus (RSV) is commonest cause of lower respiratory tract disease in children worldwide. Pathogenesis of RSV-induced bronchiolitis is poorly understood, and effects of RSV infection on ion transport (a seminal function of respiratory epithelial cells) have not been investigated. I hypothesize that RSV infection of respiratory epithelial cells reduces their Na ? transport capacity. Preliminary studies have demonstrated that this hypothesis is correct, both in vitro and in RSV-infeeted BALB/c mice. My aims for years 01- 03 are to: (1) quantify alterations in Na + transport across airway and alveolar epithelia in vivo and ex vivo, after infection of BALB/c mice with RSV; (2) define changes in Na + currents and amiloride-sensitive channel activity after RSV infection of murine epithelial cells in vitro; and (3) correlate alterations in Na + transport induced by RSV in vitro and in vivo with alterations in ENaC expression by marine respiratory epithelia. My plan for years 04-05 is to determine the role of the ubiquitin/proteasome pathway in mediating reduced Na+ transport after RSV infection of respiratory epithelia, and to identify the role of the RSV small hydrophobic (SH) gene product in modulation of Na + transport. I will use a combined electrophysiologic and biochemical approach to investigate effects of RSV on Na + transport at all levels, from the single cell to the whole animal, and to correlate these effects to Na + channel expression and degradation. My project will emphasize cross-training in diverse techniques, including short-circuit measurements across monolayers, radioisotopic ion flux studies, whole cell and single channel patch-clamp, and measurement of alveolar fluid clearance and nasal potential difference in mice. ENVIRONMENT: This SERCA, with Dr. Matalon (lung physiology) as mentor and Dr. Sullender (respiratory syncytial virus) as co-mentor, will provide the training and setting I require for my maturation into an independent scientist focused on comparative pathophysiologic effects of respiratory viruses on normal epithelial cell function.

候选人：我于1992年毕业于布里斯托尔大学，并以解剖学和兽医科学为荣誉。在UAB，我在动物模型中接受了实验室动物医学，实验动物病理学和病毒发病机理的培训（2000年2月；导师Patricia Fultz博士）。随后，我选择加入Matalon博士的研究小组，在那里我可能会在电生理学中交叉开采对病毒发病机理的了解，并应用这些技术来研究人类呼吸道病毒性肺炎的发病机理。 研究：呼吸道合胞病毒（RSV）是全球儿童下呼吸道疾病的最常见原因。 RSV诱导的细支气管炎的发病机理知之甚少，RSV感染对离子转运的影响（呼吸性上皮细胞的开创功能）尚未研究。我假设RSV感染呼吸道上皮细胞会降低其NA吗？运输能力。初步研究表明，在体外和RSV感染的BALB/C小鼠中，该假设都是正确的。我的目标多年01- 03是：（1）在感染了RSV的BALB/C小鼠后，量化了体内和体内气道和牙槽上皮的Na +转运的变化； （2）在体外RSV感染后，定义了Na +电流和Amiloride敏感的通道活性的变化； （3）与RSV在体外和体内诱导的Na +转运的变化与海洋呼吸性上皮的ENAC表达改变。我的计划04-05是确定泛素/蛋白酶体途径在介导RSV感染呼吸性上皮elia后Na +转运降低的作用，并确定RSV小疏水性（SH）基因在Na +转运模型中的作用。我将使用合并的电生理学和生化 研究RSV对从单个细胞到整个动物的所有水平上Na +转运的影响的方法，并将这些效应与Na +通道表达和降解相关联。我的项目将强调各种技术的交叉训练，包括单层之间的短路测量，放射性分离的离子通量研究，全细胞和单个通道贴剂钳，以及肺泡液清除率和小鼠鼻电势差的测量。 环境：这个SERCA，Matalon博士（肺部生理学）作为导师，Sullender博士（呼吸道合胞病毒）作为同事，将为我提供我所需的培训和设置，以使我的成熟成为一个独立的科学家，该独立科学家专注于呼吸病毒对正常上皮细胞功能的比较病理生理学作用。