Regulation of Erythrocyte Volume Homeostasis

红细胞容量稳态的调节

• 批准号: 8927631
• 负责人: PATRICK G GALLAGHER
• 金额: $ 18.73万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-15 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8927631
• 关键词: Address; Adverse effects; Anemia; Biological; Biological Assay; Cations; Cell model; Cell physiology; Cells; Dehydration; Disease; Electrophysiology (science); Endothelial Cells; Erythrocyte Aging; Erythrocyte Survival; Erythrocyte volume; Erythrocytes; Erythroid Cells; Genetic; Goals; Health; Hemoglobinopathies; Hemolytic Anemia; Homeostasis; Hydration status; In Vitro; Inborn Genetic Diseases; Inherited; Knowledge; Lead; Lymphocyte; Maintenance; Malaria; Mediating; Membrane Proteins; Modeling; Molecular; Mus; Mutation; Names; Neurons; Pathology; Pathway interactions; Patients; Permeability; Physiological; Play; Process; Proteins; Proteomics; Regulation; Role; Sickle Cell; Sickle Cell Anemia; Sodium Chloride; Stress; Stretching; Structure; Techniques; Technology; Thalassemia; Variant; Water; Work; base; cell type; human disease; in vivo; innovation; kidney cell; multidisciplinary; mutant; novel; protein expression; solute; trafficking

DESCRIPTION (provided by applicant): Maintenance of water and solute homeostasis is critical to survival of the erythrocyte. Primary disorders of erythrocyte hydration are a group of inherited disorders ranging from dehydrated to overhydrated cells. Depending on the degree of perturbation of volume homeostasis, hemolytic anemia may result. Secondary disorders of erythrocyte hydration occur when perturbation in cell hydration is associated with another condition, for instance the dehydration that commonly accompanies sickle cell disease or beta hemoglobinopathies. In secondary disorders, altered erythrocyte hydration may be a major contributor to disease pathology. PIEZO1 has recently been identified as the long sought after protein involved in mammalian mechanosensation and stretch-activated cation channel activation. We have discovered mutations in PIEZO1 that lead to hereditary xerocytosis (HX), a hemolytic anemia characterized by primary erythrocyte dehydration, indicating PIEZO1 plays an important role in cellular volume homeostasis. PIEZO1 is a candidate for unidentified stretch-induced cation pathways in the erythrocyte that play critical roles in erythrocyte aging, malaria invasion, and circulatory sheer stress. PIEZO1 is also an excellent candidate for Psickle, an unidentified cation permeability pathway in sickle erythrocytes at the initiation of the dehydratio cascade of fundamental importance to sickle cell pathobiology. Despite its importance, we have no knowledge of the mechanisms controlling PIEZO1 expression, regulation, structure or function and its role in regulation of volume homeostasis in erythroid cells. The proposed studies combine state of the art cellular, genetic, proteomic, and physiologic technologies to characterize the expression, structure and function of PIEZO1, in an innovative, multidisciplinary manner. Studies include functional, cell-based assays of PIEZO1 membrane protein expression, trafficking, and electrophysiology in a novel, in vivo stably-transfected, single-copy, inducible cll model of PIEZO1 expression. New genetically modified murine models of Piezo1, including a murine model of HX, will be created and characterized. Finally, quantitative MRM-based proteomic studies and state-of-the-art mechanotransduction physiologic techniques will be applied to erythrocytes from HX patients under a variety of cellular conditions. PIEZO1 is found in many cell types including lymphocytes, endothelial, kidney, and neural cells, indicating it likey mediates important functions in a wide variety of cells. Thus studies in erythroid cells may yield mechanistic or biological principles generalizable to many critical cellular processes or human diseases.

描述（由申请人提供）：维持水和溶质稳态对于红细胞的存活至关重要。红细胞水合的原发性疾病是一组遗传性疾病，范围从细胞脱水到过度水合。根据容量稳态的扰动程度，可能会导致溶血性贫血。当细胞水合扰动与另一种病症相关时，例如通常伴随镰状细胞病或β血红蛋白病的脱水，就会发生红细胞水合的继发性疾病。在继发性疾病中，红细胞水合作用的改变可能是疾病病理学的主要贡献者。最近，PIEZO1 被认为是长期以来备受追捧的参与哺乳动物机械感觉和拉伸激活阳离子通道激活的蛋白质。我们发现 PIEZO1 突变会导致遗传性干细胞增多症 (HX)，这是一种以原发性红细胞脱水为特征的溶血性贫血，表明 PIEZO1 在细胞体积稳态中发挥着重要作用。 PIEZO1 是红细胞中未知的拉伸诱导的阳离子途径的候选者，该途径在红细胞衰老、疟疾侵袭和循环剪切应力中发挥着关键作用。 PIEZO1 也是 Psickle 的绝佳候选者，Psickle 是镰状红细胞中一种未识别的阳离子通透性途径，在脱水级联的启动时对镰状细胞病理学至关重要。尽管它很重要，但我们对控制 PIEZO1 表达、调节、结构或功能的机制及其在红细胞体积稳态调节中的作用尚不了解。拟议的研究结合了最先进的细胞、遗传、蛋白质组和生理学技术，以创新、多学科的方式表征 PIEZO1 的表达、结构和功能。研究包括在 PIEZO1 表达的新型体内稳定转染、单拷贝、诱导性 cll 模型中对 PIEZO1 膜蛋白表达、运输和电生理学进行功能性、基于细胞的测定。将创建并表征新的 Piezo1 转基因小鼠模型，包括 HX 小鼠模型。最后，基于 MRM 的定量蛋白质组学研究和最先进的机械转导生理学技术将应用于各种细胞条件下 HX 患者的红细胞。 PIEZO1 存在于许多细胞类型中，包括淋巴细胞、内皮细胞、肾细胞和神经细胞，表明它可能在多种细胞中介导重要功能。因此，对红细胞的研究可能会产生可推广到许多关键细胞过程或人类疾病的机械或生物学原理。