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病，影响全球超过1250万人1。我们的长期目标是找到PKD的有效治疗方法。常染色体显性PKD（ADPKD）是由编码多囊蛋白1和多囊蛋白2的PKD 1和PKD 2基因突变引起的。这种疾病的特征是肾脏和肝脏中出现充满液体的囊肿，其进行性生长伴随着炎症，纤维化和代谢缺陷，通常导致慢性肾病（CKD）和终末期肾病（ESRD）。虽然我们现在有FDA批准的PKD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至关重要。该项目的目标是生成ADPKD肾脏昼夜节律破坏的初步数据，并确定治疗的靶向机制。 昼夜节律是行为和生理学中内在的周期性~24小时振荡，其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通过生物钟基因建立在细胞自主转录-翻译延迟反馈的分子机制上的。这些包括转录因子，昼夜运动输出周期kaput（CLOCK）和大脑和肌肉ARNT样蛋白-1（BMAL 1），它们驱动隐花色素（CRY）和周期（PER）基因，其产物抑制CLOCK和BMAL 1。昼夜节律调节基本的肾功能，如转运蛋白的表达、肾小管重吸收、分泌、血浆流量和肾小球滤过率。在人类糖尿病肾病、肾结石病和高血压中，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，肾功能昼夜节律被破坏。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不受控制以及盐和水的尿液排泄不平衡。已知昼夜节律破坏（时间破坏）会驱动癌症、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会导致疾病病理学。 我们进行了一项新的观察，即与对照细胞相比，PKD 1基因缺失的小鼠肾小管上皮细胞在24小时核心时钟基因（如CLOCK、BMAL 1、PER 2和BMAL 1）的昼夜节律振荡中显示出显著的破坏。与WT小鼠肾脏相比，ADPKD小鼠肾脏还显示出囊肿生长调节细胞信号传导因子的显著昼夜变化。此外，肾脏生物钟基因表达和肾脏生理昼夜节律也被发现在小鼠ADPKD肾脏中被破坏，并且与年龄和肾囊肿生长的增加相对应。重要的是，在体外研究中，使用PER 2的药理学增强剂Nobiletin的时间疗法显著降低了ADPKD细胞的细胞增殖和囊肿生长。基于这些观察，我们假设昼夜节律被破坏并促进ADPKD的疾病进展，恢复昼夜节律可以减缓或停止囊肿生长。 为了生成初步数据，将实现以下目标： 具体目标1.确定ADPKD肾脏昼夜节律破坏的潜在致病机制以及这些变化如何促进ADPKD进展。 研究将确定WT和Pkd 1 RC/RC小鼠肾脏之间时钟基因表达周期的差异。将检查脂联素-AMPK-mTOR通路作为ADPKD肾脏昼夜节律破坏的致病细胞信号传导通路的作用，并检查其与脂肪代谢的可能联系。为了确定这些变化是否有助于ADPKD进展，将在时间中断研究中检查选定的基因和细胞信号传导机制。 具体目标2。确定时间中断是否有助于ADPKD的表型改变或疾病进展。 我们建议检查肾小管特异性基因敲除BMAL 1（一个重要的时钟基因）在ADPKD小鼠中的作用。PKD 1 RC/RC-BMAL 1f/f-PkhD 1cre小鼠将通过将PKD 1 RC/RC小鼠与BMAL 1f/f小鼠和PkhD 1cre小鼠交配产生。将表征疾病进展中的年龄依赖性变化，以确定BMA 11基因缺失（时间破坏）对囊肿生长早期ADPKD进展的影响。