Role of Muc1 in the b-catenin Response to Acute Kidney Injury

Muc1 在 b-catenin 对急性肾损伤反应中的作用

基本信息

批准号：
10323751
负责人：
Mohammad Al-bataineh
金额：
$ 5.4万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-15 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10323751
关键词：
Acute Acute Renal Failure with Renal Papillary Necrosis Affect Animal Model Animals Apical Attenuated Award Binding Biological Assay Blood Career Choice Cell Line Cell Nucleus Cessation of life Chronic Chronic Kidney Failure Clinical Complex Contralateral Cytoplasm Development Disease Progression End stage renal failure Epithelial Female Gender Genetic Transcription Glycogen Synthase Kinase 3 Glycoproteins Goals Health Heterozygote Human Hypotension Infection Injury Injury to Kidney Ischemia Kidney Kidney Diseases Knock-out Knockout Mice Laboratory Research Luciferases Mediating Mentored Research Scientist Development Award Mentors Microscopic Microscopy Modeling Molecular Biology Techniques Morphology Mucin 1 protein Mus Names Nephrectomy Nuclear Outcome Oxygen Pathology Pathway interactions Patients Pharmacology Phase Phenotype Phosphorylation Physiological Physiology Play Positioning Attribute Prevention Proteins Recovery Regulation Renal function Renal tubule structure Reperfusion Injury Reporter Reporting Research Research Personnel Research Project Grants Research Training Risk Role Sepsis Severities Signal Pathway Signal Transduction Surface Technical Expertise Testing Three-Dimensional Imaging Time Tissues Training Transactivation Transfection Transgenic Organisms Translational Research Up-Regulation Weight base beta catenin career cell injury congenic design experimental study glycogen synthase kinase 3 beta in vivo Model inhibitor/antagonist injury and repair injury recovery insight kidney cell kidney fibrosis male mouse model multiphoton microscopy mutant neoplastic cell nephrogenesis novel strategies novel therapeutic intervention novel therapeutics overexpression prevent promoter renal ischemia response tenure track

项目摘要

Project summary/abstract Dr. Al-bataineh’s career goals for the award period are to develop a research project and professional expertise that will enable him to develop scientific independence from his mentor and to establish his own career path to independent translational research in acute kidney injury. Dr. Al-bataineh will further his laboratory research and training by developing his proficiency in (i) validated animal models of renal injury including the two-kidney hanging-weight mouse model of ischemia-reperfusion injury (IRI), and mouse model of AKI-CKD progression, (ii) advanced approaches of kidney microscopy including multiphoton microscopy, three-dimensional imaging, and quantitative microscopic analyses, and (iii) technical expertise in molecular biology techniques such as ChIPseq and promoter-luciferase reporter assays. Dr. Al-bataineh plans to make the transition to an independent, tenure track position within 2-3 years of his mentored research training during this award. His long-term career goal is to become a fully independent academic investigator in the broad fields of renal physiology and kidney disease, performing research that provides insight into fundamental physiological problems that impact clinical issues, with a particular focus on AKI pathology, prevention, and treatment. -catenin signaling is a complex cellular response that is activated in renal tubules during kidney injury. While moderate ischemia results in transient induction of -catenin levels that associated with kidney protection, severe ischemia leads to sustained activation of the -catenin pathway and development of kidney fibrosis, implicating severity of injury as a key determinant of long term outcome. Emerging evidence supports a role for the transmembrane glycoprotein mucin 1 (MUC1 in humans, Muc1 in animals) in regulating -catenin activity. In tumor cells, (i) MUC1 directly binds to -catenin in the cytoplasm and nucleus, (ii) MUC1 blocks glycogen synthase kinase 3 (GSK3)-mediated degradation of -catenin, and (iii) MUC1 overexpression correlates with increased levels of nuclear -catenin and its transcriptional activity. We previously reported that Muc1, found on the apical surface of normal kidney epithelia, plays a protective role in a mouse model of ischemia-reperfusion injury (IRI) by comparing kidney function and morphology in Muc1 KO mice and congenic control mice (AJP-Renal 2015; PMID: 25925251). Our recent studies showed that Muc1 induction and targeting to the nucleus after moderate ischemia was associated with increased -catenin levels and signaling (AJP-Renal 2016; PMID: 26739894). We also observed that sustained upregulation of Muc1 was also associated with prolonged induction of -catenin in mouse kidney homogenates at 7 d after severe ischemia. Based on these findings and on previous reports related to the acute and chronic effects of -catenin after kidney injury, we hypothesize that Muc1 protects against the early stages of AKI via transactivation of - catenin pathway, but persistent Muc1/-catenin signaling after severe AKI leads to CKD progression and renal fibrosis. Our proposed studies will utilize both a mouse model of moderate and severe IRI, and a cultured human kidney cell line (HEK-293) after ATP depletion as an established model of ischemic cell injury. Moreover, we have already obtained mice with varying levels of Muc1 expression (Muc1 knockout, heterozygotes, and a transgenic overexpressing human MUC1) that will be used to assess if increased levels of Muc1 in the kidney in the short term enhance protection during IRI, while prolonged expression of Muc1 promotes kidney fibrosis. Successful completion of the proposed studies will provide important insight into the signaling pathways that are altered during IRI and repair, and will provide valuable information to help us design novel therapeutic strategies to treat AKI and prevent CKD progression.

项目概要/摘要 Al-bataineh 博士在获奖期间的职业目标是开发一个研究项目和专业人士专业知识将使他能够摆脱导师的束缚，建立自己的科学独立性 Al-bataineh 博士将进一步推进他在急性肾损伤方面的独立转化研究的职业道路。通过提高他在以下方面的熟练程度进行实验室研究和培训：（i）经过验证的肾损伤动物模型包括两肾悬重小鼠缺血再灌注损伤模型、小鼠模型 AKI-CKD 进展，(ii) 肾脏显微镜检查的先进方法，包括多光子显微镜检查，三维成像和定量显微分析，以及（iii）分子技术专业知识 Al-bataineh 博士计划制定诸如 ChIPseq 和启动子荧光素酶报告基因等生物学技术。在他指导的研究培训后的 2-3 年内过渡到独立的终身教授职位他的长期职业目标是成为广泛领域中完全独立的学术研究者。肾脏生理学和肾脏疾病，进行深入了解基本原理的研究影响临床问题的生理问题，特别关注 AKI 病理学、预防和治疗 -连环蛋白信号传导是一种复杂的细胞反应，在肾脏过程中在肾小管中被激活。中度缺血会导致与肾脏相关的 β-连环蛋白水平短暂诱导。保护，严重缺血导致 β-连环蛋白途径持续激活和肾脏发育纤维化，表明损伤的严重程度是长期结果的关键决定因素。跨膜糖蛋白粘蛋白 1（人类为 MUC1，动物为 Muc1）在调节 β-连环蛋白中的作用在肿瘤细胞中，(i) MUC1 直接与细胞质和细胞核中的 β-catenin 结合，(ii) MUC1 阻断。糖原合酶激酶 3  (GSK3) 介导的 -连环蛋白降解，以及 (iii) MUC1 过表达与核 -连环蛋白水平及其转录活性的增加相关。 Muc1 存在于正常肾上皮细胞的顶端表面，在小鼠模型中发挥保护作用通过比较 Muc1 KO 小鼠和同系小鼠的肾功能和形态来观察缺血再灌注损伤 (IRI) 对照小鼠（AJP-Renal 2015；PMID：25925251）。我们最近的研究表明 Muc1 诱导和中度缺血后靶向细胞核与 β-连环蛋白水平和信号传导增加有关（AJP-Renal 2016；PMID：26739894）我们还观察到 Muc1 也持续上调。与严重缺血后 7 天小鼠肾匀浆中 β-连环蛋白的长时间诱导有关。根据这些发现以及之前有关 -连环蛋白的急性和慢性影响的报告肾损伤，我们研究 Muc1 通过 - 反式激活来预防 AKI 的早期阶段连环蛋白通路，但严重 AKI 后持续的 Muc1/-连环蛋白信号传导会导致 CKD 进展和肾脏疾病我们提出的研究将利用中度和重度 IRI 小鼠模型以及培养的小鼠模型。 ATP耗尽后的人肾细胞系（HEK-293）作为缺血性细胞损伤的既定模型。此外，我们已经获得了具有不同水平 Muc1 表达的小鼠（Muc1 敲除，杂合子和转基因过表达人类 MUC1），将用于评估水平是否增加 Muc1 在肾脏中的短期表达增强了 IRI 期间的保护，而 Muc1 的长期表达促进肾纤维化的成功完成拟议的研究将为我们提供重要的见解。在 IRI 和修复过程中发生改变的信号通路将提供有价值的信息来帮助我们设计新的治疗策略来治疗 AKI 并预防 CKD 进展。