New mouse model of cisplatin-induced AKI and development of prevention therapy

顺铂诱发 AKI 的新小鼠模型及预防治疗的进展

基本信息

批准号：
10671738
负责人：
Jianjun Zhao
金额：
$ 52.93万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10671738
关键词：
Acceleration Actins Acute Renal Failure with Renal Papillary Necrosis Address Affect Antisense Oligonucleotides Attenuated Base Excision Repairs Binding Binding Sites Biopsy Specimen Cancer Model Cancer Patient Cell Death Cell Nucleus Cells Cisplatin Clinical Clinical Trials Credentialing DNA DNA Adduction DNA Adducts DNA Damage DNA Repair Gene Data Development Disease Dose Drug usage Engineering Etiology Event Functional disorder Future Genetic Engineering Genetic Transcription Genetically Engineered Mouse Genomics Glomerular Filtration Rate Heart Human Hyperuricemia Hypokalemia Inflammatory Injury to Kidney Ischemia KRASG12D Kidney Knockout Mice Knowledge Malignant Neoplasms Malignant neoplasm of lung Maps Mediating Mediator Metabolism Minor Mitochondria Mitochondrial DNA Modeling Molecular Mus Mutation Myosin Heavy Chains Nature Nephritis Nucleotide Excision Repair Oxidative Stress Oxygen Consumption Pathogenesis Pathologic Pathology Pathway interactions Patients Phenotype Polymerase Prevention therapy Preventive Prophylactic treatment Proteins Renal function Research Project Grants Rest Role Sampling Signal Transduction Solid Neoplasm Symptoms Testing Time Tissues Toxic effect Transgenic Mice Transgenic Organisms Translating Translational Research Treatment Efficacy Tubular formation Tubulointerstitial Nephritis Up-Regulation Withdrawal adduct chemotherapy clinically relevant crosslink cytotoxic deafness design drug development endonuclease experimental study glomerulosclerosis high throughput screening homologous recombination in vivo inhibitor innovation interest knock-down mouse model neoplastic cell nephrogenesis nephrotoxicity new therapeutic target novel prevent prophylactic renal damage repair enzyme repaired response small molecule inhibitor standard care targeted treatment translational model translational study tumor

项目摘要

SUMMARY Cisplatin is among the most widely used drugs against solid tumors, but it can cause severe damage of renal tubulointerstitial tissues, manifesting itself pathologically in the form of glomerular filtration rate reduction, hype- ruricemia, hypokalemia, and accelerated secondary glomerulosclerosis and glomerular ischemia. The end re- sult is acute kidney injury (AKI). Cisplatin induces AKI, which afflicts 25% of cisplatin-treated cancer patients worldwide. With few exceptions, there is no effective preventive or post-exposure therapy for cisplatin-induced AKI (C-AKI). Oxidative stress and mitochondrial damage are drivers of AKI-associated pathology; however, the molecular pathways that mediate these events are poorly defined. Using murine C-AKI model kidney and bi- opsy samples from C-AKI patients, we have recently found that both oxidative stress and mitochondrial dam- age are associated with upregulation of renal apurinic/apyrimidinic endonuclease 2 (APE2). In preliminary ex- periments, we discovered that cisplatin exposure in vivo increases expression of APE2 in mice and inhibits the action of the protein myosin heavy chain 9 (MYH9), a key regulatory molecule of kidney function. These altera- tions translate into impact on tubulointerstitial tissue. Our overall working hypothesis is that cisplatin alters kidney APE2 signaling, leading to inhibition of MYH9 with concomitant tubulointerstitial damage. Here we pro- pose to more precisely illuminate the target role of APE2 in AKI development, and specifically hypothesize (i) that highly expressed APE2 affects mtDNA integrity and MYH9/actin intermediate mitochondrial fission and (ii) that this in turn promotes renal injury by triggering cell-death pathways. To address these dual issues, we propose using APE2 transgenic and knockout (KO) mouse models to pursue the two Specific Aims detailed below. In Aim 1, we will validate a genetically engineered murine model of APE2 with direct clinical relevance to C-AKI. We will comprehensively define the APE2 mouse phenotype and compare it with that of the C-AKI mouse model and the human C-AKI signature with respect to pathophysiological features at the molecular level using whole genomic, transcriptional, and immunomic approaches. We will define the binding site of APE2/MYH9 and its functional significance for further translational studies. In Aim 2, we will conduct proof-of- concept studies on APE2 targeted therapy for prevention of C-AKI. Additionally, we will define a novel targeted therapy to prevent nephrotoxicity by delivering APE2 gapmer antisense oligonucleotides or small-molecule in- hibitors to selectively suppress APE2 expression in proximal tubule cells in APE2 transgenic and C-AKI mice. Impact: Accomplishing these Specific Aims will be the first step toward constructing a temporal-mechanistic map of events that occur upon cisplatin exposure and translate into tubulointerstitial nephritis and may ulti- mately enable development of novel targeted therapy for AKI disease.

概括顺铂是针对实体瘤的最广泛使用的药物之一，但可能会导致肾脏严重损害肾小管间质组织，以肾小球滤过速率降低，炒作形式表现出自身的表现罗西血症，低钾血症和加速的继发性肾小球硬化和肾小球缺血。终点重新 Sult是急性肾脏损伤（AKI）。顺铂诱导AKI，折磨于25％的顺铂治疗癌症患者全世界。除少数例外，没有有效的预防或暴露后治疗顺铂诱导 Aki（C-Aki）。氧化应激和线粒体损伤是AKI相关病理学的驱动因素。但是，介导这些事件的分子途径的定义很差。使用鼠C-Aki模型肾脏和Bi- 来自C-Aki患者的Opsy样品，我们最近发现氧化应激和线粒体大坝既是年龄与肾脏肾上腺素/肾上腺素核酸内切酶2（APE2）的上调有关。在初步内部，我们发现在体内的顺铂暴露会增加小鼠APE2的表达，并抑制蛋白质肌球蛋白重链9（MYH9）的作用，这是肾功能的关键调节分子。这些Altera- tions转化为对微管间质组织的影响。我们的整体工作假设是顺铂改变了肾脏APE2信号传导，导致抑制MyH9并伴随的微管间质损伤。在这里我们亲摆姿势更精确地阐明APE2在AKI开发中的目标作用，并特别假设（i）高度表达的APE2影响mtDNA完整性和MyH9/actin中间线粒体裂变和（ii）这反过来又通过触发细胞死亡途径来促进肾脏损伤。为了解决这些双重问题，我们建议使用APE2转基因和基因敲除（KO）鼠标模型追求两个特定目的详细以下。在AIM 1中，我们将验证具有直接临床相关性的基因设计的APE2的基因工程鼠模型到C-Aki。我们将全面定义APE2鼠标表型，并将其与C-Aki的表型进行比较小鼠模型和人类C-AKI签名相对于分子的病理生理特征使用全基因组，转录和免疫学方法的水平。我们将定义约束位点 APE2/MYH9及其对进一步翻译研究的功能意义。在AIM 2中，我们将进行证明 APE2靶向预防C-AKI的概念研究。此外，我们将定义一个针对性的新颖通过递送APE2 Gapmer反义寡核苷酸或小分子IN-的疗法来预防肾毒性在APE2转基因和C-AKI小鼠中选择性抑制近端小管细胞中APE2表达的吉他。影响：实现这些特定目标将是构建时间机械的第一步顺铂暴露时发生的事件图，并转化为肾小管肾炎，可能最终促使人们能够开发针对AKI疾病的新型靶向疗法。