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

• 批准号: 10440020
• 负责人: Mohammad Al-bataineh
• 金额: $ 6.75万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 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/10440020
• 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

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 博士Al-bataineh在获奖期间的职业目标是开发一个研究项目和专业 专业知识，使他能够发展科学独立于他的导师，并建立自己的 在急性肾损伤的独立转化研究的职业道路。Al-bataineh博士将进一步 实验室研究和培训，提高他在（i）经验证的肾损伤动物模型方面的熟练程度 包括两肾悬重缺血再灌注损伤（IRI）小鼠模型和 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镜分析，以及（iii）分子生物学方面的技术专长 生物学技术如ChIPseq和启动子-荧光素酶报告基因测定。艾尔-巴塔尼博士计划 过渡到一个独立的，终身职位在2-3年内，他的指导研究培训期间 这个奖。他的长期职业目标是成为广泛领域的完全独立的学术研究者 肾脏生理学和肾脏疾病，进行研究，提供洞察基础 影响临床问题的生理问题，特别关注阿基病理学、预防和 治疗β-连环蛋白信号传导是一种复杂的细胞反应，在肾脏疾病期间在肾小管中被激活。 损伤而中度缺血导致短暂的β-catenin水平的诱导， 保护，严重缺血导致β-连环蛋白通路的持续激活和肾脏的发育 纤维化，暗示损伤的严重程度是长期结果的关键决定因素。新出现的证据支持 跨膜糖蛋白粘蛋白1（人MUC 1，动物Muc 1）在调节β-连环蛋白中的作用 活动在肿瘤细胞中，（i）MUC 1直接结合细胞质和细胞核中的β-连环蛋白，（ii）MUC 1阻断肿瘤细胞中的β-连环蛋白。 糖原合成酶激酶3 β（GSK 3 β）介导的β-连环蛋白降解，和（iii）MUC 1过表达 与核β-连环蛋白及其转录活性水平的增加相关。我们之前报道过， 在正常肾上皮细胞的顶面发现的Muc 1，在一个小鼠模型中起保护作用。 缺血再灌注损伤（IRI），通过比较Muc 1 KO小鼠和同类基因小鼠的肾功能和形态学 对照小鼠（AJP-Renal 2015; PMID：25925251）。我们最近的研究表明，Muc 1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的增加有关。 （AJP-Renal 2016; PMID：26739894）。我们还观察到，Muc 1的持续上调也是 与严重缺血后7天小鼠肾匀浆中β-连环蛋白的诱导延长有关。 基于这些发现和以前的报告，涉及急性和慢性影响的β-连环蛋白后， 肾损伤时，我们假设Muc 1通过转录激活阿基-1， 连环蛋白通路，但严重阿基后持续的Muc 1/β-连环蛋白信号传导导致CKD进展和肾损害。 纤维化我们提出的研究将利用中度和重度IRI的小鼠模型，以及培养的 人肾细胞系（HEK-293）在ATP耗尽后作为缺血性细胞损伤的建立模型。 此外，我们已经获得了具有不同Muc 1表达水平的小鼠（Muc 1敲除， 杂合子和过表达人MUC 1的转基因），其将用于评估是否增加的水平 短期内肾脏中Muc 1的表达增强了IRI期间的保护作用，而长期表达Muc 1 促进肾纤维化成功完成拟议的研究将提供重要的洞察力， 在IRI和修复过程中改变的信号通路，并将提供有价值的信息，以帮助我们 设计新的治疗策略来治疗阿基和预防CKD进展。