The role of Sirtuin 5 in acute kidney injury

Sirtuin 5在急性肾损伤中的作用

• 批准号: 10618353
• 负责人: Sunder Sims-Lucas
• 金额: $ 45.55万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10618353
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Affect; Age; Animal Model; Apoptosis; Automobile Driving; Cells; Cellular Metabolic Process; Chemicals; Chronic; Chronic Kidney Failure; Cisplatin; Clinical; Coupled; Data; Data Correlations; Disease model; Dose; Enzymes; Epithelial Cells; Exhibits; Functional disorder; Goals; Histologic; Hospitalization; Human; Human Cell Line; In Vitro; Infusion procedures; Injury; Injury to Kidney; Ischemia; Kidney; Knock-out; Knockout Mice; Life Expectancy; Link; Liver; Long-Chain-Acyl-CoA Dehydrogenase; Lysine; Measures; Metabolic; Mitochondria; Modeling; Morbidity - disease rate; Mouse Cell Line; Mus; Organelles; Oxidative Stress; Pathogenesis; Pathology; Pathway interactions; Patients; Peptides; Proteins; Proteomics; Reactive Oxygen Species; Renal function; Reperfusion Injury; Reperfusion Therapy; Repression; Resistance; Risk; Role; Sirtuins; Site; Small Interfering RNA; Testing; Therapeutic; Tissues; Tubular formation; Up-Regulation; Western Blotting; acyl-CoA oxidase; comparison control; effective therapy; enzyme activity; fatty acid oxidation; genetic manipulation; in vivo; in vivo Model; knock-down; metabolomics; mitochondrial metabolism; mortality; nephrotoxicity; novel; novel therapeutics; oxidative damage; peroxisome; pharmacologic; prevent; renal ischemia; repaired; response; therapeutic target

ABSTRACT Acute kidney injury (AKI) occurs in nearly 1 of 5 hospitalized patients and is associated with increased morbidity and mortality across all ages. Many AKI patients will recover kidney function post-injury but then progress to chronic kidney disease (CKD). The mechanisms are poorly understood and there are currently no effective therapies to prevent, limit, or reverse the tissue damage. There is a critical need to identify mechanisms involved in the pathogenesis of AKI. Our long-term goal is to elucidate these mechanisms and leverage them for new therapies to limit AKI and prevent the transition to CKD. Proximal tubule epithelial cells (PTEC), a major site of damage during AKI, are very metabolically active and rich in mitochondria. Mitochondrial metabolism causes increased reactive oxygen species (ROS) which has been implicated in both ischemia-reperfusion injury (IRI) and cisplatin-induced nephrotoxicity. Modulating mitochondrial function during AKI is an attractive, but thus far unachievable, strategy. Our central hypothesis is that loss of the mitochondrial sirtuin lysine deacylase Sirt5 leads to shifts in PTEC metabolism that protects against AKI. This is supported by preliminary data showing protection against both IRI and cisplatin-induced AKI in global Sirt5 knockout (Sirt5-/-, Sirt5+/-) mice in vivo and in vitro as well as in primary human PTEC with siRNA knockdown of Sirt5. Further data support our proposed mechanism of protection in which Sirt5-/- PTEC exhibit a form of metabolic adaptation characterized by a shift of fatty acid oxidation (FAO) from mitochondria to peroxisomes. Peroxisomes have previously been linked to renoprotection in other animal models, most likely due to their ability to eliminate ROS. In Sirt5-/- kidneys, peroxisomes are more resistant to damage during AKI. Our central hypothesis will be tested with two aims. Aim 1 will define the specific site of Sirt5 action during kidney injury with a particular focus on PTEC. While aim 2 will drill down on the renoprotective role of metabolic FAO inhibition coupled with stimulation of peroxisomal fatty acid oxidation during kidney injury. Both aims will utilize a rigorous, mechanistic approach that combines in vitro and in vivo models. In vivo studies in mice will use both global Sirt5-/- PTEC-specific knockout of Sirt5 as well as global and PTEC-specific knockout of LCAD-/- (key mitochondrial FAO enzyme). In vitro studies will use isolated primary mouse and human PTEC as well as genetically manipulated mouse and human cell lines. Human AKI will be modeled in mice by unilateral ischemia-reperfusion injury and single high dose treatment with the nephrotoxin cisplatin. We have also optimized a CKD model using a unilateral ischemia-reperfusion injury model. This project will significantly advance the field by opening up new therapeutic avenues where Sirt5 can be pharmacologically inhibited or its renoprotective mechanism can be harnessed in the context of AKI to protect against injury and block the progression to chronic kidney disease.

抽象的 近五分之一的住院患者发生急性肾损伤 (AKI)，并且与发病率增加相关 以及各个年龄段的死亡率。许多 AKI 患者在受伤后会恢复肾功能，但随后会进展为 慢性肾脏病（CKD）。人们对其机制知之甚少，目前尚无有效的方法 预防、限制或逆转组织损伤的疗法。迫切需要确定所涉及的机制 AKI 的发病机制。我们的长期目标是阐明这些机制并利用它们开发新的 限制 AKI 并防止转变为 CKD 的疗法。近端小管上皮细胞（PTEC）是 AKI 期间的损伤，代谢非常活跃并且富含线粒体。线粒体代谢的原因 活性氧（ROS）增加，与缺血再灌注损伤（IRI）有关 和顺铂引起的肾毒性。在 AKI 期间调节线粒体功能是一种很有吸引力的方法，但到目前为止 无法实现的，战略。我们的中心假设是线粒体 Sirtuin 赖氨酸脱酰酶 Sirt5 的缺失 导致 PTEC 代谢发生变化，从而预防 AKI。初步数据表明这一点 对整体 Sirt5 敲除 (Sirt5-/-, Sirt5+/-) 小鼠体内 IRI 和顺铂诱导的 AKI 的保护作用 在体外以及通过 siRNA 敲低 Sirt5 的原代人 PTEC 中。进一步的数据支持我们的建议 Sirt5-/- PTEC 表现出一种以转变为特征的代谢适应形式的保护机制 从线粒体到过氧化物酶体的脂肪酸氧化（FAO）。过氧化物酶体以前被认为与 在其他动物模型中具有肾脏保护作用，很可能是由于它们消除 ROS 的能力。在 Sirt5-/- 肾脏中， 过氧化物酶体在 AKI 期间更能抵抗损伤。我们的中心假设将通过两个目标进行检验。目的 图 1 将定义肾损伤期间 Sirt5 作用的具体位点，特别关注 PTEC。虽然目标 2 将 深入研究代谢性FAO抑制与过氧化物酶体脂肪刺激的肾脏保护作用 肾损伤期间的酸氧化。这两个目标都将采用严格的机械方法，结合体外 和体内模型。小鼠体内研究也将使用 Sirt5 的全局 Sirt5-/- PTEC 特异性敲除 LCAD-/-（关键线粒体FAO酶）的全局和PTEC特异性敲除。体外研究将使用 分离原代小鼠和人类 PTEC 以及基因操纵的小鼠和人类细胞系。 将通过单侧缺血再灌注损伤和单次高剂量治疗在小鼠中建立人类 AKI 模型 肾毒素顺铂。我们还使用单侧缺血再灌注损伤优化了 CKD 模型 模型。该项目将开辟新的治疗途径，Sirt5 可以显着推进该领域的发展 可以通过药物抑制或在 AKI 背景下利用其肾脏保护机制来保护 防止损伤并阻止慢性肾脏疾病的进展。

项目成果

Dicarboxylic Acid Dietary Supplementation Protects against AKI.

膳食补充剂二羧酸可预防 AKI。

• DOI: 10.1681/asn.0000000000000266
• 发表时间: 2024
• 期刊: Journal of the American Society of Nephrology : JASN
• 作者: SilvaBarbosa,AnneC;Pfister,KatherineE;Chiba,Takuto;Bons,Joanna;Rose,JacobP;Burton,JordanB;King,ChristinaD;O'Broin,Amy;Young,Victoria;Zhang,Bob;Sivakama,Bharathi;Schmidt,AlexandraV;Uhlean,Rebecca;Oda,Akira;Schilling,Birg
• 通讯作者: Schilling,Birg