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

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.

慢性肾病 (CKD) 影响着近 15% 的美国人，肾损伤通常针对肾小管 上皮细胞。这些肾小管如何反应可以决定肾脏是否进行修复或肾小管间质修复 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病如何 肾损伤会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 肾小管的存活和 TIF 的发展。众所周知，细胞周期、新陈代谢和线粒体 功能都是紧密协调的过程，但尚不清楚上皮G1到S细胞周期如何进展 影响 CKD 肾脏的代谢。初步数据表明，减少 G1 期的细胞周期进程 肾小管中的 S 期可防止啮齿类 CKD 模型中的纤维化并减少肾小管细胞凋亡。在 此外，减少 G1 到 S 的进程会增加葡萄糖氧化，即葡萄糖代谢为丙酮酸 然后通过柠檬酸循环和电子传递链在线粒体中被氧化。这 该提案将检验以下假设：减少 CKD 中上皮 G1 期进展至 S 期可保护 通过改变代谢来对抗上皮损伤和纤维化。为了测试这一点，目标 1 将使用 药物（palbociclib）或遗传（有条件地删除肾小管中的细胞周期蛋白 D1）方法来减少 G1 小鼠 S 细胞周期进展。我们假设减少上皮细胞 G1 期向 S 期的进展 细胞通过减少肾小管损伤和纤维化在 CKD 模型中具有保护作用。我们的初步数据表明 减少受损肾组织和孤立肾小管细胞中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。该目标旨在研究如何 减少细胞周期进展可能会改变这些信号传导途径，从而减少肾小管损伤和肌成纤维细胞损伤 分别通过自分泌和旁分泌信号激活。第二个目标是研究代谢 使用 Seahorse 生物通量减少 G1 期至 S 期进展的受损肾小管中发生的变化 分析仪、14C-丙酮酸离体氧化研究和稳定同位素代谢组学。我们假设 减少上皮细胞周期进程增加葡萄糖氧化，从而改善上皮细胞存活率 减少纤维化，部分是通过 AMP 激活的蛋白激酶途径实现的。我们还将研究葡萄糖如何 肾小管的氧化与代谢无关，影响对慢性损伤的反应。细胞的影响 线粒体功能和结构的循环进展也将使用 Oroboros 和 super- 分辨率显微镜。这些研究应该提供关于上皮细胞如何变化的新信息 循环和代谢影响对慢性肾损伤的反应，并有可能鉴定新的 治疗 CKD 的治疗目标。