Cell Cycle and Metabolism in Chronically Injured Renal Tubules

慢性损伤肾小管的细胞周期和代谢

• 批准号: 10661066
• 负责人: Leslie S Gewin
• 金额: $ 45.34万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-13 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661066
• 关键词: 5' -AMP-activated protein kinase; Affect; American; Apoptosis; Autocrine Communication; CDK4 gene; Catalytic Domain; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Respiration; Cells; Cellular Metabolic Process; Chronic; Chronic Kidney Failure; Citric Acid Cycle; Cyclin D1; Cyclin-Dependent Kinases; Cyclins; DNA biosynthesis; Data; Development; Disease model; Electron Transport; Environment; Enzymes; Epithelial Cells; Epithelium; FDA approved; Fibrosis; G2/M Arrest; Genetic; Genus Hippocampus; Glucose; Glycolysis; Hypoxia; Inflammation; Inflammatory; Injury; Injury to Kidney; Kidney; Kidney Diseases; Knockout Mice; Lactic acid; Link; Mediating; Metabolic; Metabolism; Mitochondria; Mitosis; Mus; Myofibroblast; Natural regeneration; Nutrient; Paracrine Communication; Pathway interactions; Pharmaceutical Preparations; Phenotype; Play; Process; Proliferating; Pyruvate; Renal tubule structure; Resolution; Rodent; Role; S phase; Signal Pathway; Signal Transduction; Structure; Testing; Tissues; Tubular formation; anaerobic glycolysis; cardiovascular disorder risk; conditional knockout; cytokine; epithelial injury; epithelial repair; fatty acid oxidation; glucose metabolism; improved; in vitro activity; in vivo; injured; innovation; kidney fibrosis; kidney metabolism; metabolomics; mouse model; new therapeutic target; novel; oxidation; pharmacologic; preclinical study; repaired; response; response to injury; stable isotope; superresolution microscopy; tool; 5' -AMP-activated protein kinase; Affect; American; Apoptosis; Autocrine Communication; CDK4 gene; Catalytic Domain; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Respiration; Cells; Cellular Metabolic Process; Chronic; Chronic Kidney Failure; Citric Acid Cycle; Cyclin D1; Cyclin-Dependent Kinases; Cyclins; DNA biosynthesis; Data; Development; Disease model; Electron Transport; Environment; Enzymes; Epithelial Cells; Epithelium; FDA approved; Fibrosis; G2/M Arrest; Genetic; Genus Hippocampus; Glucose; Glycolysis; Hypoxia; Inflammation; Inflammatory; Injury; Injury to Kidney; Kidney; Kidney Diseases; Knockout Mice; Lactic acid; Link; Mediating; Metabolic; Metabolism; Mitochondria; Mitosis; Mus; Myofibroblast; Natural regeneration; Nutrient; Paracrine Communication; Pathway interactions; Pharmaceutical Preparations; Phenotype; Play; Process; Proliferating; Pyruvate; Renal tubule structure; Resolution; Rodent; Role; S phase; Signal Pathway; Signal Transduction; Structure; Testing; Tissues; Tubular formation; anaerobic glycolysis; cardiovascular disorder risk; conditional knockout; cytokine; epithelial injury; epithelial repair; fatty acid oxidation; glucose metabolism; improved; in vitro activity; in vivo; injured; innovation; kidney fibrosis; kidney metabolism; metabolomics; mouse model; new therapeutic target; novel; oxidation; pharmacologic; preclinical study; repaired; response; response to injury; stable isotope; superresolution microscopy; tool

Chronic kidney disease (CKD) affects almost 15% of Americans, and renal injury often targets the renal tubule epithelia. How these tubules respond can determine whether the kidney undergoes repair or tubulointerstitial fibrosis (TIF), the common hallmark of progressive CKD. This proposal focuses on understanding how chronic renal injury induces changes in the renal tubular cell cycle and metabolism and how these changes affect tubular survival and the development of TIF. It is well known that cell cycle, metabolism, and mitochondrial function are all closely coordinated processes, but it is not clear how epithelial G1 to S cell cycle progression affects metabolism in the CKD kidney. Preliminary data suggests that reducing cell cycle progression from G1 to S phase in renal tubules protects against fibrosis in rodent CKD models and decreases tubular apoptosis. In addition, reducing G1 to S progression increased glucose oxidation, the metabolism of glucose to pyruvate which is then oxidized in the mitochondria through the citric acid cycle and electron transport chain. This proposal will test the hypothesis that reducing epithelial G1 to S phase progression in CKD protects against epithelial injury and fibrosis through altered metabolism. To test this, Aim 1 will use either a pharmacologic (palbociclib) or a genetic (conditionally delete cyclin D1 in renal tubules) approach to reduce G1 to S cell cycle progression in mice. We hypothesize that decreasing G1 progression to S phase in epithelial cells is protective in CKD models by reducing tubular injury and fibrosis. Our preliminary data show that reducing cell cycle progression in both injured kidney tissue and in isolated tubule cells also suppresses signaling pathways and inflammatory cytokines associated with kidney injury. This aim investigates how reducing cell cycle progression may alter these signaling pathways to reduce tubule injury and myofibroblast activation by autocrine and paracrine signaling, respectively. The second aim investigates the metabolic changes that occur in injured tubules with reduced G1 to S phase progression using the Seahorse bioflux analyzer, 14C-pyruvate oxidation studies ex vivo, and stable isotopic metabolomics. We hypothesize that reducing epithelial cell cycle progression increases glucose oxidation leading to better epithelial survival and less fibrosis, in part, through the AMP-activated protein kinase pathway. We will also investigate how glucose oxidation in renal tubules, independent of metabolism, affects the response to chronic injury. The impact of cell cycle progression on mitochondrial function and structure will also be defined using Oroboros and super- resolution microscopy. These studies should provide novel information about how changes in epithelial cell cycle and metabolism affect the response to chronic renal injury with the potential identification of novel therapeutic targets to treat CKD.

慢性肾脏疾病（CKD）影响了几乎15％的美国人，肾脏损伤通常针对肾小管 上皮。这些小管的反应如何确定肾脏是进行修复还是微管间隙 纤维化（TIF），是进行性CKD的共同标志。该提案的重点是了解慢性 肾损伤引起肾小管细胞周期和代谢的变化，以及这些变化如何影响 管状存活和TIF的发展。众所周知，细胞周期，代谢和线粒体 功能都是紧密协调的过程，但尚不清楚上皮G1到S细胞周期的进展如何 影响CKD肾脏中的新陈代谢。初步数据表明，减少了G1细胞周期的进程 肾小管中的S相可防止啮齿动物CKD模型中的纤维化并减少管状细胞凋亡。在 此外，将G1降低到S进展增加了葡萄糖氧化，葡萄糖代谢为丙酮酸 然后通过柠檬酸周期和电子传输链在线粒体中氧化。这 提案将检验以下假设：将上皮G1降低到CKD中的S相进展 通过改变新陈代谢来抵抗上皮损伤和纤维化。为了测试这一点，AIM 1将使用 药理学（PALBOCICLIB）或遗传（肾小管中有条件删除细胞周期蛋白D1）方法可减少G1 小鼠细胞周期的进展。我们假设降低了上皮中的G1进展到S相位 通过减少管状损伤和纤维化，细胞在CKD模型中具有保护性。我们的初步数据表明 降低受伤肾脏组织和孤立小管细胞的细胞周期进程也抑制 与肾损伤相关的信号通路和炎症细胞因子。这个目标调查了如何 减少细胞周期的进程可能会改变这些信号通路以减少小管损伤和肌纤维细胞 自分泌和旁分泌信号传导的激活。第二个目标调查了代谢 使用Seahorse BioFlux降低G1到S相进展的受伤小管中发生的变化 分析仪，14C-丙酮酸氧化研究，体内稳定的同位素代谢组学。我们假设这一点 减少上皮细胞周期进程会增加葡萄糖氧化，从而获得更好的上皮存活和 部分通过AMP激活的蛋白激酶途径的部分纤维化。我们还将研究葡萄糖 与代谢无关的肾小管中的氧化会影响对慢性损伤的反应。细胞的影响 线粒体功能和结构上的循环进展也将使用Oroboros和Super-定义 分辨率显微镜。这些研究应提供有关上皮细胞如何变化的新信息 循环和代谢影响对慢性肾脏损伤的反应，并具有新的鉴定 治疗CKD的治疗靶标。