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 1,250 万人1。我们的长期目标是找到治疗 PKD 的有效疗法。常染色体显性 PKD (ADPKD) 是由 PKD1 和 PKD2 基因突变引起的。编码多囊蛋白 1 和多囊蛋白 2。该疾病的特点是肾脏和肝脏中出现充满液体的囊肿，囊肿的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 批准的治疗 PKD 的药物，但开发更好的 ADPKD 疗法和生活方式改变策略至关重要。该项目的目标是生成 ADPKD 肾脏昼夜节律紊乱的初步数据，并确定治疗的目标机制。 昼夜节律是行为和生理学中内在的 24 小时周期性振荡，它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建立在时钟基因的细胞自主转录翻译延迟反馈分子机制的基础上。驱动 Cryptochrome 的转录因子、昼夜节律运动输出周期 kaput (CLOCK) 以及大脑和肌肉 ARNT 样蛋白 1 (BMAL1) (CRY) 和 period (PER) 基因，其产物抑制 CLOCK 和 BMAL1，调节基本肾功能，例如转运蛋白的表达、肾小管重吸收、分泌、血浆流量和肾小球滤过率。糖尿病肾脏中的肾功能昼夜节律被破坏。人类疾病、肾结石疾病和高血压，以及腺嘌呤诱导的 CKD 和顺铂诱导的急性肾损伤的小鼠模型中，重要的是，基因突变或缺失。小鼠肾脏中的时钟基因会导致血压不受控制以及盐和水的尿液排泄不平衡，已知昼夜节律紊乱（时间紊乱）会导致癌症、代谢综合征、肝脏疾病和阿尔茨海默病的进展，但目前尚不清楚是否会导致这种情况。 ADPKD 的昼夜节律紊乱会导致疾病病理。 我们进行了一项新的观察，与对照细胞相比，具有 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 小时昼夜节律振荡显着受到破坏，而 ADPKD 小鼠肾脏也显示出显着的昼夜变化。与WT小鼠肾脏相比，囊肿生长调节细胞信号因子此外，还发现了肾脏生物钟基因表达和肾脏生理昼夜节律。重要的是，在体外研究中，使用 PER2 药理学增强剂 Nobiletin 进行的时间疗法显着减少了 ADPKD 细胞的细胞增殖和囊肿生长。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阻止囊肿生长。 为了生成初步数据，将实现以下目标： 具体目标 1. 确定 ADPKD 肾脏昼夜节律紊乱的致病机制以及这些变化如何导致 ADPKD 进展。 研究将确定 WT 和 Pkd1RC/RC 小鼠肾脏之间时钟基因表达周期性的差异。脂联素-AMPK-mTOR 通路的作用将作为 ADPKD 肾脏昼夜节律破坏的致病细胞信号通路进行检查，并可能与此相关。为了确定这些变化是否会导致 ADPKD 进展，将在时间紊乱研究中检查选定的基因和细胞信号传导机制。 具体目标 2. 确定时间紊乱是否会导致 ADPKD 的表型变化或疾病进展。 我们建议通过将 PKD1RC/RC 小鼠与 BMAL1f/f 小鼠交配来检测 ADPKD 小鼠中 BMAL1（一种重要的时钟基因）的肾小管特异性基因敲除的效果。和 PkhD1cre 小鼠的疾病进展的年龄依赖性变化将被表征，以确定 BMAl1 基因缺失（时间紊乱）对 ADPKD 的影响。囊肿生长期间的早期进展。