Bipartite regulation of cellular osmosensing in C. elegans

线虫细胞渗透感应的双向调节

基本信息

批准号：
8630544
负责人：
SAMUEL T LAMITINA
金额：
$ 9.17万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8630544
关键词：
Address Age Age Factors Aging Aging-Related Process Animal Model Animals Bacteria Biological Assay Biology Buffers Caenorhabditis elegans Caenorhabditis elegans Proteins Cell Culture Techniques Cell Volumes Cell physiology Cells Chronic Kidney Failure Complex Crowding Cultured Cells Cytoskeleton Data Defect Detection Diabetic Neuropathies Diagnosis Disease Early Diagnosis Early Intervention Environment Exhibits Extracellular Matrix Extracellular Matrix Proteins Failure Gene Expression Genes Genetic Homeostasis Human Hypertension Immune response Integral Membrane Protein Kidney Kidney Diseases Kidney Failure Knowledge Lead Life Mammalian Cell Mechanics Mediating Membrane Modeling Molecular Mucins Names Osmoregulation Pathway interactions Peripheral Nervous System Diseases Physiological Physiological Processes Physiology Play Process Property Proteins Quality Control Regulation Role Signal Pathway Signal Transduction Stress Stress-Induced Protein Stretching Subcutaneous Tissue System Testing Tissues Water Yeasts age related base biological adaptation to stress blood pressure regulation cell growth regulation genome wide association study heat shock transcription factor in vivo insight mutant novel therapeutic intervention polyglutamine positional cloning programs protein aggregation protein misfolding protein structure public health relevance response sensor solute stressor urinary

项目摘要

Bipartite regulation of cellular osmosensing in C. elegans The physiological process of maintaining cellular solute and water content is termed osmotic homeostasis, or osmoregulation, and is essential for all forms of cellular life. In humans, osmotic homeostasis plays vital roles in several contexts, including regulation of the kidney's urinary concentrating mechanism, control of blood pressure, and activation of immune responses. Osmotic dyshomeostasis is associated with several age- related diseases, including chronic kidney disease, renal failure, hypertension, and peripheral neuropathy. Despite the obvious importance of osmoregulation in both physiological and pathophysiological disease states, little is known about the mechanisms by which animal cells sense and respond to osmotic stress. A better understanding of these mechanisms may allow earlier detection and intervention in age-related diseases. Most studies of osmoregulation have been carried out using cultured cells, which fail to mimic the complex environments in which most cells are found. These studies have led to many hypotheses to explain the mechanism(s) of cellular osmosensing, such as mechanical 'stretching' of the membrane and/or cytoskeleton, macromolecular crowding, and alterations in cytoplasmic ionic content, to name a few. However, there is little data supporting any of these models. To gain an in vivo perspective on mechanisms of cellular osmosensing in animals, we are studying this process in the model organism C. elegans, in which complex cell-cell and cell- extracellular matrix (ECM) interactions are preserved. Using unbiased forward and reverse genetic approaches, we discovered critical roles for the extracellular matrix (Rohlfing et al, PLoS Genetics, 2011) and protein misfolding (Moronetti Mazzeo et al, PNAS, 2012) in the regulation of cellular osmosensing in C. elegans. Based on these findings we hypothesize that animal cells use both mechanotransduction and protein damage detection mechanisms to sense osmotic disturbances and activate osmosensitive gene expression. In Aim 1, we will determine if the C. elegans cuticular ECM acts as a structural 'osmosensor' to transduce information via interactions between the mucin-like protein OSM-8 and a transmembrane protein PTR-23. In Aim 2, we will define the native proteins susceptible to stress-induced protein aggregation and determine how aging and aging regulators influence osmotic stress induced protein damage and osmosensitive gene expression. In Aim 3, we will examine how ECM and protein damage detection pathways interact with each other to control osmoregulatory physiology. Our studies take maximal advantage of the C. elegans system to fill an important gap in our knowledge of metazoan cell physiology. These findings will provide transformative insights into the conserved process of osmoregulation that will allow us to better understand, detect, and manage age-related diseases of osmotic dyshomeostasis.

线虫细胞渗透感应的双向调节维持细胞溶质和水含量的生理过程称为渗透稳态，或渗透调节，对于所有形式的细胞生命都至关重要。在人类中，渗透稳态起着至关重要的作用在多种情况下，包括调节肾脏的尿液浓缩机制、控制血液压力和免疫反应的激活。渗透压失调与多种年龄相关相关疾病，包括慢性肾病、肾功能衰竭、高血压和周围神经病变。尽管渗透调节在生理和病理生理疾病状态中都具有明显的重要性，人们对动物细胞感知和响应渗透压的机制知之甚少。更好的了解这些机制可能有助于更早地发现和干预与年龄相关的疾病。大多数渗透调节研究都是使用培养细胞进行的，这些细胞无法模拟复杂的渗透调节过程。大多数细胞存在的环境。这些研究提出了许多假设来解释细胞渗透传感的机制，例如膜和/或细胞骨架的机械“拉伸”，仅举几例，大分子拥挤和细胞质离子含量的改变。然而，却很少有支持任何这些模型的数据。获得细胞渗透传感机制的体内视角在动物中，我们正在模型生物秀丽隐杆线虫中研究这一过程，其中复杂的细胞-细胞和细胞- 细胞外基质（ECM）相互作用得以保留。使用无偏正向和反向遗传方法，我们发现了细胞外基质的关键作用（Rohlfing 等人，PLoS Genetics，2011）和蛋白质错误折叠（Moronetti Mazzeo 等人，PNAS，2012）在 C. 线虫。基于这些发现，我们假设动物细胞同时使用机械转导和蛋白质损伤检测机制来感知渗透紊乱并激活渗透敏感基因表达。在目标 1 中，我们将确定线虫表皮 ECM 是否充当结构“渗透传感器”来转导通过粘蛋白样蛋白 OSM-8 和跨膜蛋白 PTR-23 之间的相互作用来获取信息。在目标 2，我们将定义易受应激诱导的蛋白质聚集影响的天然蛋白质，并确定如何衰老和衰老调节因子影响渗透应激诱导的蛋白质损伤和渗透敏感基因表达。在目标 3 中，我们将研究 ECM 和蛋白质损伤检测途径如何相互作用其他控制渗透调节生理机能。我们的研究最大限度地利用了秀丽隐杆线虫系统填补了我们关于后生动物细胞生理学知识的一个重要空白。这些发现将提供变革性的对渗透调节保守过程的深入了解将使我们能够更好地理解、检测和管理与年龄相关的渗透失调疾病。