Circadian Clock Disruption in the Pathogenesis and Therapy of Polycystic Kidney Disease

多囊肾病发病机制和治疗中的昼夜节律紊乱

• 批准号: 10475900
• 负责人: Reena Rao
• 金额: $ 10万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2023-03-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10475900
• 关键词: ARNT gene; ARNTL gene; Acute Renal Failure with Renal Papillary Necrosis; Adenine; Adult; Affect; Age; Alzheimer' s Disease; Autosomal Dominant Polycystic Kidney; Behavior; Biological Process; Blood Pressure; Brain; Breeding; Cell Proliferation; Cells; Chronic Kidney Failure; Chronotherapy; Circadian Dysregulation; Circadian Rhythms; Cisplatin; Combined Modality Therapy; Cyst; Cystic kidney; Data; Defect; Development; Diabetic Nephropathy; Disease; Disease Progression; Diurnal Rhythm; End stage renal failure; Enhancers; Epithelial Cells; Excretory function; FDA approved; FRAP1 gene; Feedback; Fibrosis; Gene Deletion; Gene Expression; Gene Mutation; Genes; Genetic Transcription; Glomerular Filtration Rate; Goals; Growth; Hour; Human; Hypertension; In Vitro; Inflammation; Inherited; Kidney; Kidney Calculi; Kidney Diseases; Life Style Modification; Light; Link; Liquid substance; Liver; Liver diseases; Magnetic Resonance Imaging; Malignant Neoplasms; Metabolic; Metabolic syndrome; Molecular; Motor Activity; Mus; Muscle; Mutation; Output; PKD1 gene; PKD2 gene; PKD2 protein; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Patients; Periodicity; Pharmaceutical Preparations; Pharmacology; Phase; Phenotype; Physiological; Physiology; Plasma; Polycystic Kidney Diseases; Proteins; Regimen; Renal function; Renal tubule structure; Reporter; Role; Signal Pathway; Signal Transduction; Sodium Chloride; System; Testing; Time; Time-restricted feeding; Translations; Tubular formation; Water; adiponectin; age related; base; circadian; circadian pacemaker; cryptochrome; effective therapy; feeding; knockout gene; lipid metabolism; mouse model; nobiletin; novel; novel therapeutics; polycystic kidney disease 1 protein; restoration; shift work; targeted treatment; tolvaptan; transcription factor; transcriptome; treatment strategy; urinary

SPECIFIC AIMS: Polycystic Kidney Disease (PKD) is the most common inherited kidney disease that affects over 12.5 million people worldwide 1. Our long-term goal is to find effective therapies for PKD. Autosomal dominant PKD (ADPKD) is caused by mutations of PKD1 and PKD2 genes that encode for polycystin 1 and polycystin 2. The disease features development of fluid-filled cysts in the kidneys and liver, the progressive growth of which is accompanied by inflammation, fibrosis and metabolic defects, often leading to chronic kidney disease (CKD) and end stage renal disease (ESRD). Although we now have an FDA approved drug for PKD, it is critically important to develop better therapies and lifestyle modification strategies for ADPKD patients. The goal of this project is to generate preliminary data on circadian rhythm disruption in ADPKD kidneys, and identify mechanisms to target for therapy. Circadian rhythms are intrinsic cyclical ~24-hour oscillations in behavior and physiology that coordinate the diverse biological processes with the time of day. The mammalian circadian system is built upon a cell-autonomous transcription-translation delayed feedback molecular mechanism by the clock genes. These include the transcription factors, circadian locomotor output cycles kaput (CLOCK) and brain and muscle ARNT-like protein-1 (BMAL1), which drive Cryptochrome (CRY) and Period (PER) genes, whose products inhibit CLOCK and BMAL1. Circadian rhythms regulate fundamental renal functions such as expression of transporters, tubular reabsorption, secretion, plasma flow and glomerular filtration rate. Renal functional circadian rhythms are disrupted in diabetic kidney disease, kidney stone disease and hypertension in humans, and in mouse models of adenine induced CKD and cisplatin induced acute kidney injury. Importantly, gene mutation or deletion of clock genes in mouse kidneys result in uncontrolled blood pressure and unbalanced urinary excretion of salt and water. Circadian rhythm disruption (chronodisruption) is known to drive disease progression in cancer, metabolic syndrome, liver diseases and Alzheimer’s disease. However, it is currently unknown if circadian rhythm disruptions in ADPKD contribute to disease pathology. We made a novel observation that mouse tubular epithelial cells with PKD1 gene deletion show significant disruption in 24h circadian oscillations of core clock genes such as CLOCK, BMAL1, PER2 and CRY1, when compared to control cells. ADPKD mouse kidneys also showed significant diurnal variations in cyst-growth regulating cell signaling factors compared to WT mouse kidneys. Moreover, renal circadian clock gene expression and renal physiological diurnal rhythms were also found to be disrupted in mouse ADPKD kidneys and corresponded to increase in age and renal cyst growth. Importantly, chronotherapy using Nobiletin, a pharmacological enhancer of PER2 significantly reduced cell proliferation and cyst growth by ADPKD cells in in vitro studies. Based on these observations, we hypothesized that circadian rhythms are disrupted and promote disease progression in ADPKD and restoring the circadian rhythm can slow or stop cyst growth. To generate preliminary data, the following aims will be accomplished: Specific Aim 1. To determine pathogenic mechanisms underlying disruption of circadian rhythms in ADPKD kidneys and how such changes contribute to ADPKD progression. Studies will identify differences in periodicity of expression of clock genes between WT and Pkd1RC/RC mouse kidneys. The role of adiponectin-AMPK-mTOR pathway will be examined as a pathogenic cell signaling pathway for circadian disruption in the ADPKD kidney, and possible links to fat metabolism will be examined. To determine if such changes contribute ADPKD progression, selected genes and cell signaling mechanisms will be examined in chronodisruption studies. Specific Aim 2. To determine if chronodisruption contributes to phenotypic change or disease progression in ADPKD. We propose to examine the effect of renal tubule-specific gene knockout of BMAL1, (an important clock gene) in ADPKD mice. PKD1RC/RC-BMAL1f/f-PkhD1cre mouse will be generated by breeding PKD1RC/RC mice with BMAL1f/f mice and PkhD1cre mice. Age dependent changes in disease progression will be characterized to determine the effect of BMAl1 gene deletion (chronodisruption) on ADPKD progression early during cyst growth.

具体目标： 多囊肾病(PKD)是最常见的遗传性肾脏疾病，影响全球超过1250万人1。我们的长期目标是找到有效的PKD治疗方法。常染色体显性遗传性PKD(ADPKD)是由编码多囊蛋白1和多囊蛋白2的PKD1和PKD2基因突变引起的。该病的特征是在肾脏和肝脏形成充满液体的囊肿，其进行性增长伴随着炎症、纤维化和代谢缺陷，常导致慢性肾脏疾病(CKD)和终末期肾脏疾病(ESRD)。虽然我们现在有FDA批准的治疗PKD的药物，但为ADPKD患者开发更好的治疗方法和生活方式改变策略是至关重要的。该项目的目标是生成有关ADPKD肾脏昼夜节律紊乱的初步数据，并确定靶向治疗的机制。 昼夜节律是行为和生理中内在的周期性~24小时的振荡，协调不同的生物过程与一天中的时间。哺乳动物的昼夜节律系统是由时钟基因建立在细胞自主转录-翻译延迟反馈分子机制之上的。这些包括转录因子，昼夜运动输出周期kaput(CLOCK)和脑和肌肉Arnt-like Protein-1(BMAL1)，它们驱动隐色素(CRY)和周期(PER)基因，其产物抑制CLOCK和BMAL1。昼夜节律调节基本的肾功能，如转运蛋白的表达、肾小管的重吸收、分泌、血浆流量和肾小球滤过率。在人类的糖尿病肾病、肾结石疾病和高血压中，以及在腺嘌呤诱导的CKD和顺铂诱导的急性肾损伤的小鼠模型中，肾功能昼夜节律被打乱。重要的是，小鼠肾脏中时钟基因的基因突变或缺失会导致血压失控和尿盐和水排出不平衡。众所周知，昼夜节律紊乱(时序紊乱)会导致癌症、代谢综合征、肝病和阿尔茨海默病的疾病进展。然而，目前尚不清楚ADPKD的昼夜节律紊乱是否与疾病病理有关。 我们做了一个新的观察，与对照细胞相比，PKD1基因缺失的小鼠肾小管上皮细胞的核心时钟基因如Clock、BMAL1、PER2和CRY1的24小时昼夜振荡明显中断。与WT小鼠肾脏相比，ADPKD小鼠肾脏也显示出显著的囊性生长调节细胞信号因子的日变化。此外，ADPKD小鼠肾脏的生物钟基因表达和肾脏生理昼夜节律也被打乱，并与年龄增加和肾囊肿生长相对应。重要的是，在体外研究中，使用PER2的药物增强剂诺比莱汀的计时疗法显著减少了ADPKD细胞的细胞增殖和囊泡生长。基于这些观察，我们假设昼夜节律被打乱并促进ADPKD的疾病进展，恢复昼夜节律可以减缓或阻止囊肿的生长。 为了生成初步数据，将实现以下目标： 具体目的1.确定ADPKD肾脏昼夜节律紊乱的发病机制，以及这种变化如何促进ADPKD的进展。 研究将确定WT和Pkd1RC/RC小鼠肾脏时钟基因表达周期的差异。脂联素-AMPK-mTOR通路的作用将作为ADPKD肾脏昼夜节律紊乱的致病细胞信号通路进行研究，并将研究与脂肪代谢的可能联系。为了确定这些变化是否有助于ADPKD的进展，将在时序中断研究中检查选定的基因和细胞信号机制。 具体目的2.确定时序紊乱是否与ADPKD的表型改变或疾病进展有关。 我们建议在ADPKD小鼠中检测肾小管特异性基因BMAL1(一个重要的时钟基因)的敲除效果。将PKD1RC/RC小鼠与BMAL1f/f小鼠和PkhD1cre小鼠杂交，产生PKD1RC/RC-BMAL1f/f-PkhD1cre小鼠。疾病进展中与年龄相关的变化将被用来确定BMal1基因缺失(时序中断)在囊性生长早期对ADPKD进展的影响。