Acid secretion and intracellular pH regulation in proximal-tubule cells

近曲小管细胞的酸分泌和细胞内 pH 调节

• 批准号: 9766272
• 负责人: Rossana Occhipinti
• 金额: $ 10.82万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-18 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9766272
• 关键词: 3-Dimensional; Acid-Base Equilibrium; Acidosis; Acids; Affect; Apical; Area; Bicarbonates; Blood; Boron; Buffers; Carbon Dioxide; Carbonic Anhydrase II; Cell membrane; Cell model; Cells; Clinical; Computer Simulation; Computers; Data; Dependence; Development; Development Plans; Diffusion; Disease; Dyes; Electrodes; Electrophysiology (science); Elements; Environmental air flow; Epithelial Cells; Epithelium; Generations; Goals; Heart; In Vitro; Ion-Selective Electrodes; Ions; Kidney; Kinetics; Knowledge; Laboratories; Life; Liquid substance; Literature; Lung; Maintenance; Measurement; Measures; Membrane; Mentored Research Scientist Development Award; Mentors; Mentorship; Methods; Modeling; Molecular Biology; Monitor; Movement; Mus; Nephrons; Oocytes; Optics; Organ; Perfusion; Pharmacology; Physiological; Physiology; Play; Positioning Attribute; Preparation; Process; Property; Proteins; Proton Pump; Proximal Kidney Tubules; Reaction; Recovery; Regulation; Renal tubular acidosis; Renal tubule structure; Research; Research Personnel; Role; Side; Site; Sodium Bicarbonate; Surface; Testing; Time; Training; Urine; Work; Writing; Xenopus oocyte; apical membrane; base; basolateral membrane; carbonate dehydratase; career; career development; complex biological systems; digital imaging; experience; experimental study; extracellular; fluorescence imaging; insight; mathematical model; multi-scale modeling; multidisciplinary; novel; optical imaging; post-doctoral training; preservation; professor; public health relevance; response; sensor; skills; sound; therapeutic target; tool; vector; virtual; voltage clamp

 DESCRIPTION (provided by applicant): The K01 award will facilitate the mentored transition of Dr. Rossana Occhipinti from a postdoctoral position to an independent investigator, capable of combining in-silico (i.e., using computer) and in-vitro experiments in the field of multi-scale modeling of kidney acid-base physiology. Dr. Occhipinti is an applied mathematician with expertise in mathematical modeling of complex biological systems and acid-base physiology. Her postdoctoral training has been in the wet-laboratory of her primary mentor, Professor Walter Boron. Here, her research has focused on developing sophisticated mathematical models to simulate the data on intracellular pH (pHi) and extracellular-surface pH obtained in the laboratory from electrophysiology experiments. However, because Dr. Occhipinti's research has focused only on mathematical modeling and her career goals depend on having expertise in two distinct but complementary areas, it is critical that, in addition to perform state-of-art modeling she acquires the knowledge and ability to perform sound in-vitro experiments to inform her in-silico experiments. This K01 award will provide Dr. Occhipinti with the necessary protected amount of time to obtain appropriate training in electrophysiology, proximal tubule (PT) perfusion, and optics while continuing to expand her mathematical modeling expertise. Dr. Occhipinti's career development plan includes focused coursework, mentorship from a multi-disciplinary group of leading investigators, and practical research experience focusing on training in: (1) molecular biology, transport/electrophysiology, and oocyte experiments (Aim 1); (2) renal physiology and proximal- tubule perfusion/optical imaging (Aim 2); and (3) sophisticated mathematical modeling (Aim 3). Moreover, Dr. Occhipinti will engage in professional development and networking activities. The research plan focuses on the unknown key question of how proximal-tubule cells can reconcile the competing demand of two vital processes: H+ secretion versus regulation of their own pHi. To answer this question, the research plan has 3 aims that will assess key properties of the major acid-base transporters in PTs: (1) the electrogenic Na/HCO3 cotransporter (NBCe1-A) at the basolateral membrane (BLM), (2) the Na-H exchanger 1 (NHE1) at the BLM, and (3) the NHE3 at the apical membrane. In Aim 1, Dr. Occhipinti will determine for the first time the pHi dependence of NBCe1-A as heterologously expressed in oocytes. In Aim 2, she will assess the roles of NHE1, NHE3 and NBCe1-A from pHi recording on intact PTs. In Aim 3 she will use the results from Aim 1 and Aim 2 to create and inform a comprehensive and novel mathematical model of acid-base transport and pHi regulation for a PT cell. By pursuing these specific aims, Dr. Occhipinti will develop the knowledge and skills to perform electrophysiology and optical experiments that will (1) allow her to be independent of other laboratories in gathering the data needed to test and validate her mathematical models, and (2) prepare her to write her first R01 application. Dr Occhipinti life's work should inform our understanding of the many clinical conditions that challenge or interfere with the ability of the kidney to handle acid loads, and should be applicable in a general sense to a wide range of other epithelia.

 描述（由申请人提供）：K 01奖项将促进Rossana Occhipinti博士从博士后职位转变为独立研究者，能够结合计算机模拟（即，使用计算机）和肾脏酸碱生理学的多尺度建模领域中的体外实验。Occhipinti博士是一位应用数学家，擅长复杂生物系统和酸碱生理学的数学建模。她的博士后培训一直在她的主要导师沃尔特·博龙教授的湿实验室进行。在这里，她的研究重点是开发复杂的数学模型，以模拟从电生理学实验中获得的细胞内pH值（pHi）和细胞外表面pH值的数据。然而，由于Occhipinti博士的研究只专注于数学建模，她的职业目标取决于在两个不同但互补的领域拥有专业知识，因此，除了进行最先进的建模外，她还必须获得进行合理的体外实验的知识和能力，以告知她的计算机实验。该K 01奖项将为Occhipinti博士提供必要的保护时间，以获得电生理学，近端小管（PT）灌注和光学方面的适当培训，同时继续扩展她的数学建模专业知识。Occhipinti博士的职业发展计划包括重点课程，来自领先研究人员的多学科小组的指导，以及专注于以下培训的实践研究经验：（1）分子生物学，运输/电生理学和卵母细胞实验（目标1）;（2）肾生理学和近端小管灌注/光学成像（目标2）;以及（3）复杂的数学建模（目标3）。此外，Occhipinti博士将从事专业发展和网络活动。该研究计划的重点是未知的关键问题，即近端小管细胞如何协调两个重要过程的竞争需求：H+分泌与自身pHi的调节。为了回答这个问题，研究计划有3个目标，将评估PT中主要酸碱转运蛋白的关键特性：（1）基底外侧膜（BLM）的产电Na/HCO 3协同转运蛋白（NBCe 1-A），（2）BLM的Na-H交换剂1（NHE 1），（3）顶膜的NHE 3。在目标1中，Occhipinti博士将首次确定NBCe 1-A在卵母细胞中异源表达的pHi依赖性。在目标2中，她将从完整PT的pHi记录中评估NHE 1、NHE 3和NBCe 1-A的作用。在目标3中，她将使用目标1和目标2的结果来创建和通知PT细胞的酸碱转运和pHi调节的全面和新颖的数学模型。通过追求这些特定目标，Occhipinti博士将发展进行电生理学和光学实验的知识和技能，这些实验将（1）使她能够独立于其他实验室收集测试和验证她的数学模型所需的数据，（2）为她编写第一个R 01应用程序做好准备。Occhipinti博士的工作应该告知我们对许多挑战或干扰肾脏处理酸负荷能力的临床条件的理解，并且应该在一般意义上适用于广泛的其他上皮细胞。