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：
10297669
负责人：
Jianjun Zhao
金额：
$ 56.06万
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/10297669
关键词：
Actins Acute Renal Failure with Renal Papillary Necrosis Address Affect Antisense Oligonucleotides Attenuated Base Excision Repairs Binding Binding Sites Cancer Model Cancer Patient Cell Death Cell Nucleus Cells Cisplatin Clinical Clinical Trials DNA DNA Adducts DNA Damage DNA Repair 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 Knock-out Knockout Mice Knowledge Malignant Neoplasms Malignant neoplasm of lung Maps Mediating Mediator of activation protein Metabolism Minor Mitochondria Mitochondrial DNA Modeling Molecular Mus Mutation Myosin Heavy Chains Myosin S-2 Nature Nephritis Nucleotide Excision Repair Oxidative Stress Oxygen Consumption Partner in relationship 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 TP53 gene Testing Time Tissues Toxic effect Transgenic Mice Transgenic Organisms Translating Translational Research Treatment Efficacy Tubular formation Tubulointerstitial Nephritis Up-Regulation chemotherapy clinically relevant crosslink cytotoxic deafness design drug development endonuclease glomerulosclerosis high throughput screening homologous recombination in vivo inhibitor/antagonist innovation interest knock-down mouse model neoplastic cell nephrogenesis nephrotoxicity new therapeutic target novel overexpression prevent prophylactic renal damage repaired response small molecule 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.

概括顺铂是最广泛使用的抗实体瘤药物之一，但它会导致严重的肾损伤肾小管间质组织，病理上表现为肾小球滤过率降低，尿酸血症、低钾血症以及加速继发性肾小球硬化和肾小球缺血。结局重新结果是急性肾损伤（AKI）。顺铂可诱发 AKI，25% 的接受顺铂治疗的癌症患者患有 AKI 全世界。除少数例外，对于顺铂引起的感染没有有效的预防或暴露后治疗。 AKI（C-AKI）。氧化应激和线粒体损伤是 AKI 相关病理的驱动因素；然而，介导这些事件的分子途径尚不清楚。使用小鼠 C-AKI 模型肾脏和双通过对 C-AKI 患者的检查样本，我们最近发现氧化应激和线粒体损伤年龄与肾脱嘌呤/脱嘧啶核酸内切酶 2 (APE2) 的上调有关。在初步的前在实验中，我们发现体内顺铂暴露会增加小鼠中 APE2 的表达并抑制蛋白质肌球蛋白重链 9 (MYH9) 的作用，是肾功能的关键调节分子。这些改变这些变化转化为对肾小管间质组织的影响。我们的总体工作假设是顺铂改变肾 APE2 信号传导，导致 MYH9 抑制并伴随肾小管间质损伤。在这里我们亲为了更精确地阐明 APE2 在 AKI 发展中的目标作用，并具体假设 (i) 高表达的 APE2 影响 mtDNA 完整性和 MYH9/肌动蛋白中间线粒体裂变，（ii）这反过来又通过触发细胞死亡途径促进肾损伤。为了解决这些双重问题，我们建议使用 APE2 转基因和敲除 (KO) 小鼠模型来追求详细的两个具体目标以下。在目标 1 中，我们将验证具有直接临床相关性的 APE2 基因工程小鼠模型至 C-AKI。我们将全面定义 APE2 小鼠表型，并将其与 C-AKI 小鼠表型进行比较小鼠模型和人类 C-AKI 特征在分子水平上的病理生理学特征使用全基因组、转录和免疫组学方法。我们将定义结合位点 APE2/MYH9 及其对进一步转化研究的功能意义。在目标 2 中，我们将进行证明- APE2 靶向治疗预防 C-AKI 的概念研究。此外，我们将定义一个新颖的目标通过递送 APE2 gapmer 反义寡核苷酸或小分子来预防肾毒性的治疗抑制剂选择性抑制 APE2 转基因小鼠和 C-AKI 小鼠近曲小管细胞中 APE2 的表达。影响：实现这些具体目标将是构建时间机制的第一步顺铂暴露后发生的事件图并转化为肾小管间质性肾炎，并可能最终有力地促进了针对 AKI 疾病的新型靶向治疗的开发。