Ceramides as Novel Mediators of Tubular Metabolic Dysfunction Driving Kidney Injury

神经酰胺作为肾小管代谢功能障碍驱动肾损伤的新型调节剂

• 批准号: 10677394
• 负责人: REBEKAH JOY NICHOLSON
• 金额: $ 3.6万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677394
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Address; Aerobic; Albumins; Anabolism; Animals; Apoptosis; Apoptotic; Automobile Driving; Bilateral; Binding; Bioenergetics; Cell Culture Techniques; Cells; Ceramides; Characteristics; Chronic Disease; Chronic Kidney Failure; Clinical; Data; Data Correlations; Development; Diabetic Nephropathy; Disease; Disease Outcome; Disease Progression; Disease model; Doxycycline; Economic Burden; Electron Transport; Enzymes; Epithelial Cells; Epithelium; Evaluation; Event; Exposure to; Fatty Acids; Fellowship; Filtration; Functional disorder; Future; Genes; Genetic; Glomerular Filtration Rate; Goals; Healthcare; Histopathology; Human; Impairment; In Vitro; Inflammatory; Injury; Injury to Kidney; Intervention; Investigation; Kidney; Kidney Diseases; Knowledge; Lipids; LoxP-flanked allele; Mass Spectrum Analysis; Mediating; Mediator; Metabolic; Metabolic Diseases; Metabolic dysfunction; Metabolism; Methods; Mitochondria; Modeling; Morbidity - disease rate; Morphology; Mus; Nephrology; Non-Insulin-Dependent Diabetes Mellitus; Nonesterified Fatty Acids; Oxidative Stress; Oxygen Consumption; Pathogenesis; Pathology; Pathway interactions; Patient Care; Patients; Play; Pre-Clinical Model; Principal Investigator; Process; Production; Proximal Kidney Tubules; Reactive Oxygen Species; Renal Tissue; Renal function; Renal tubule structure; Reperfusion Injury; Reperfusion Therapy; Reporting; Research; Respiration; Risk; Rodent; Role; Running; Scientist; Sphingolipids; System; Technical Expertise; Testing; Therapeutic; Tissues; Training; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Tubular formation; Ureteral obstruction; Work; absorption; career; cell type; combat; cytokine; driving force; fatty acid oxidation; human model; impaired driving performance; improved; inhibitor; injured; insight; kidney dysfunction; kidney fibrosis; kidney metabolism; lipid metabolism; mitochondrial dysfunction; mitochondrial metabolism; mortality; mouse model; nephrogenesis; new therapeutic target; novel; novel therapeutic intervention; overexpression; pre-doctoral; preclinical study; preference; prevent; renal ischemia; serine palmitoyltransferase; single-cell RNA sequencing; solute; stable isotope; therapeutic candidate; therapeutic target; uptake

PROJECT SUMMARY/ABSTRACT. Acute kidney injury (AKI) is a prevalent condition which elicits an enormous burden on patient mortality and healthcare spending. A significant unmet need exists to elucidate kidney-specific insults driving the onset and progression of kidney injury in order to develop novel therapeutic strategies capable of directly targeting renal pathology. Compelling evidence has suggested that altered metabolism within the kidney proximal tubule is implicated in AKI. The work proposed in this fellowship application will critically evaluate the role of a class of lipotoxic lipid species, termed ceramides, as drivers of mitochondrial dysfunction, oxidative stress, and lipid accumulation characteristic of multiple kidney pathologies. Human correlational data suggest that altered tubular ceramide metabolism correlates with clinical kidney disease endpoints, such as kidney fibrosis and estimated glomerular filtration rate. Previous reports indicate that renal ceramides are elevated in pre-clinical models following kidney injury or in the setting of chronic disease, and preliminary data presented herein demonstrate that whole-body depletion of ceramides successfully prevents acute kidney injury and histopathology following renal ischemia reperfusion or obstructive injury. This application intends to address the remaining gap in knowledge regarding whether local kidney ceramides play a role in disease mechanisms and in which cell types. Specifically, the proposed project will determine if ceramides are candidate mediators of metabolic dysfunction in tubular epithelial cells. In Aim One, we will probe for protection from kidney injury incurred in novel mouse lines with genetic depletion of ceramides in the kidney tubular epithelium. Furthermore, animals with genetic gain-of-ceramide within kidney tubules will be assessed for development of kidney dysfunction and histopathology. Studies proposed in Aim Two will employ in vitro and ex vivo methods to characterize novel mechanisms of ceramides driving impairment of mitochondrial bioenergetics and lipid accumulation in kidney tubules and primary cells. Preliminary findings demonstrate that accumulation of ceramides impairs mitochondrial respiration and ATP production in cultured immortalized proximal tubular epithelial cells. This work will be the first investigation to directly assess if tubule-derived ceramides are implicated in the metabolic perturbations preceding tubular injury and histopathology and will provide valuable insight into the therapeutic potential of ceramide-lowering interventions for dominant causes of acute kidney injury. Furthermore, completion of the proposed studies will greatly enrich the applicant’s pre-doctoral training, mastery of technical skills (e.g., implementation and evaluation of kidney injury and disease models, evaluation of mitochondrial metabolism and substrate utilization, and mass spectrometry-based analysis of ceramide levels), and development as a young scientist pursuing an independent research career.

项目摘要/摘要 急性肾损伤 (AKI) 是一种普遍存在的疾病，会引发巨大的后果。 患者死亡率和医疗保健支出的负担存在着阐明肾脏特异性的重大未满足的需求。 损伤驱动肾损伤的发生和进展，以开发新的治疗策略 直接针对肾脏病理学的令人信服的证据表明，体内的代谢。 肾近曲小管与 AKI 相关。本奖学金申请中提出的工作将严格评估。 一类脂毒性脂质（称为神经酰胺）作为线粒体功能障碍、氧化 人类相关数据表明，压力和脂质积累是多种肾脏病理的特征。 改变肾小管神经酰胺代谢与临床肾脏疾病终点相关，例如肾病 纤维化和估计的肾小球滤过率先前的报告表明肾神经酰胺升高。 肾损伤后或慢性疾病背景下的临床前模型，以及提供的初步数据 本文证明，全身消耗神经酰胺可成功预防急性肾损伤， 本申请旨在解决肾缺血再灌注或阻塞性损伤后的组织病理学问题。 关于局部肾神经酰胺是否在疾病机制中发挥作用的知识仍然存在差距 具体来说，拟议的项目将确定神经酰胺是否是细胞类型的候选介质。 在目标一中，我们将探讨肾损伤的保护作用。 始于肾小管上皮神经酰胺基因缺失的新型小鼠品系。此外， 肾小管内具有神经酰胺遗传增益的动物将接受肾脏发育评估 目标二中提出的研究将采用体外和离体方法来进行功能障碍和组织病理学研究。 表征神经酰胺驱动线粒体生物能和脂质损伤的新机制 初步研究结果表明，肾小管和原代细胞中的积累。 神经酰胺损害培养的永生化近端肾小管中的线粒体呼吸和 ATP 产生 这项工作将是第一个直接评估肾小管来源的神经酰胺是否存在的研究。 与肾小管损伤和组织病理学之前的代谢紊乱有关，并将提供有价值的信息 深入了解降低神经酰胺干预措施对急性肾病主要原因的治疗潜力 此外，完成拟议的研究将极大地丰富申请人的博士前培训， 掌握技术技能（例如肾损伤和疾病模型的实施和评估、评估 线粒体代谢和底物利用的研究，以及基于质谱的神经酰胺分析 水平），以及作为一名追求独立研究事业的年轻科学家的发展。