Meprins-Metalloproteinases of the Kidney and Intestine

Meprins-肾脏和肠道金属蛋白酶

基本信息

批准号：
7920715
负责人：
JUDITH S BOND
金额：
$ 12.31万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-21 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7920715
关键词：
Acute Affect Apical Apoptotic Bacteria Bacterial Pili Basement membrane Binding Bladder Bone Marrow Cells Cell Death Cell Survival Cell surface Cells Cleaved cell Complex Cyclic AMP-Dependent Protein Kinases Cytosol Defensins Diagnosis Disease Epidermal Growth Factor Receptor Epithelial Cells Epitopes Fibronectins Functional disorder Funding Genotype Green Fluorescent Proteins Health Host Defense Hydrolysis Hypoxia Immunoprecipitation Infection Inflammatory Injury Intervention Intestines Ion Channel Protein Ischemia Kidney Knockout Mice Knowledge Lead Leukocytes Location MDCK cell Mass Spectrum Analysis Membrane Meprin Metalloproteases Molecular Morbidity - disease rate Movement Mus Necrosis Nidogen Pathogenesis Peptide Hydrolases Peptide antibodies Peptides Predisposition Prevention Protein Isoforms Proteins Proteomics Proximal Kidney Tubules Radiation Recombinants Renal tubule structure Reperfusion Injury Reperfusion Therapy Research Resources Role Severities Site Source Techniques Tissues Transfection Urinary tract infection Urologic Diseases Work apical membrane beta-Defensins brush border membrane cytokine extracellular insight irradiation kidney cell kidney epithelial cell kidney infection meprin alpha prevent research study response

项目摘要

DESCRIPTION (provided by applicant): The overall long-term objectives of this research are to elucidate mechanisms by which cell surface metalloproteinases and their secreted counterparts are regulated and interact, activate and degrade peptides and proteins at the cell surface and extracellularly, as well as to define the roles of these proteases in health and diseases such as kidney and urinary tract disease. The work has led to the discovery and characterization of meprins, complex and unique mammalian metalloproteinases abundantly expressed at the brush border membrane of kidney and intestinal epithelial cells, and in leukocytes under certain conditions. The hypothesis is that meprins have a protective role in host defense in urinary tract infections, but that the alpha/beta form is actively involved in the pathophysiology of acute renal injury. In the next period, the Specific Aims are to: (1) Determine whether meprin alpha/beta is damaging to specific proteins in kidney epithelial cells in response to hypoxia or ischemia-reperfusion. Immunohistochemical, immunoprecipitation, and proteomic techniques will be used with kidneys and proximal tubules from wild-type and meprin knockout mice to identify specific targets of meprin interaction and hydrolysis in brush border membrane, cytosol and extracellular compartments. (2) Determine whether leukocytic meprins affect movement and activity of these inflammatory cells to ischemic kidney and enhance damage. Bone marrow cells from wild-type and meprin knockout mice will be destroyed by radiation, and replaced by donor leukocytes from wild-type or meprin knockout mice carrying a green- fluorescent protein. The movement of the donor leukocytes to kidneys in the chimeric mice subjected to ischemia-reperfusion, and the injury caused by meprin positive and negative leukocytes will give insight into the damage caused by these cells. (3) Determine whether meprin alpha secreted from the kidney, or leukocytic meprins, affect the establishment and severity of urinary tract disease. Mice of different meprin genotypes will be infected with uropathic bacteria in the bladder, and the susceptibility to bladder and kidney infections will be determined. The roles of leukocytic and kidney meprins will be assessed using the chimeric mice. The activation of meprins at sites of infection, concentrations of active beta-defensins, and interactions of meprin isoforms with bacteria will be assessed to gain insights into mechanisms of protection. The availability of the meprin alpha and beta knockout mice are unique resources for these studies, and fundamental knowledge of these metalloproteinases will lead to new concepts, treatments, and interventions for kidney and urinary tract diseases.

描述（由申请人提供）：这项研究的总体长期目标是阐明机制，通过这些机制，细胞表面金属蛋白酶及其分泌的对应物受到调节和相互作用，激活和降解在细胞表面，细胞外肽和蛋白质，以及定义这些蛋白酶在健康和疾病中的作用。这项工作导致了Meprins的发现和表征，在肾脏和肠上皮细胞的刷子边界膜上以及在某些条件下的白细胞中，大量表达了复杂而独特的哺乳动物金属蛋白酶。假设是Meprins在尿路感染中具有保护作用，但α/β形式积极参与急性肾脏损伤的病理生理学。在下一个时期，具体的目的是：（1）确定MEPRINα/β是否会响应缺氧或缺血 - 再灌注而对肾上皮细胞中的特定蛋白质损害。免疫组织化学，免疫沉淀和蛋白质组学技术将与野生型和Meprin敲除小鼠的肾脏和近端小管一起使用，以鉴定MEPRIN相互作用的特定靶标和刷子边界膜，细胞质和细胞外隔室中的特定靶标。（2）确定白细胞meprins是否影响这些炎症细胞对缺血性肾脏的运动和活性并增强损伤。来自野生型和Meprin敲除小鼠的骨髓细胞将被辐射破坏，并由野生型或Meprin敲除携带绿色荧光蛋白的供体白细胞取代。供体白细胞向经历缺血 - 再灌注的嵌合小鼠中的肾脏运动，由Meprin阳性和阴性白细胞造成的损伤将使这些细胞造成的损害。（3）确定Meprin Alpha是否从肾脏分泌或白细胞Meprins会影响尿路疾病的建立和严重程度。不同MEPRIN基因型的小鼠将在膀胱中感染尿道细菌，并确定对膀胱和肾脏感染的敏感性。白细胞和肾脏Meprins的作用将使用嵌合小鼠进行评估。将评估MEPRIN在感染部位，活性β-防御素的浓度以及MEPRIN同工型与细菌的相互作用的激活，以洞悉对保护机制的见解。 Meprin Alpha和Beta淘汰小鼠的可用性是这些研究的独特资源，对这些金属蛋白酶的基本知识将导致肾脏和尿路疾病的新概念，治疗和干预措施。