Acute inhibition of renal gene expression to prevent nephrotoxicity.

急性抑制肾脏基因表达以防止肾毒性。

• 批准号: 9531353
• 负责人: Michael Thomas Eadon
• 金额: $ 15.67万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9531353
• 关键词: Acute; Acute Renal Failure with Renal Papillary Necrosis; Address; Affect; Animal Model; Appointment; Award; Biological Markers; Biopsy; Candidate Disease Gene; Caring; Cells; Cisplatin; Clinic; Clinical; Clinical Pharmacology; Clinical Trials; Clinical and Translational Science Awards; Clofarabine; Colistin; Databases; Development; Drug Modelings; Drug toxicity; Drug usage; Environment; Equilibrium; Expenditure; Exposure to; Failure; Fellowship; Foundations; Funding; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Databases; Genetic Markers; Genetic Models; Genome; Gentamicins; Glomerulonephritis; Goals; Grant; Head; Human; Indiana; Injury; Injury to Kidney; Intervention; Kidney; Kidney Diseases; Knockout Mice; LDL-Receptor Related Protein 2; Laboratories; Lasers; Learning; Learning Skill; Length of Stay; Life; Malignant Neoplasms; Medical; Membrane; Mentors; Methylation; Microdissection; Modeling; Molecular; Mus; Nephrology; Nephrons; Outcome; POU2F1 gene; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacogenomics; Phase; Phase I Clinical Trials; Phase II Clinical Trials; Physicians; Population Genetics; Premedication; Prevention; Prevention strategy; Proteins; Proximal Kidney Tubules; Recording of previous events; Research Personnel; Role; Saline; Schools; Scientist; Site; Small Interfering RNA; TP53 gene; Techniques; Testing; Therapeutic; Time; Tissues; Toxic effect; Training; Translating; Translations; Tubular formation; United States National Institutes of Health; Universities; Validation; analytical method; clinical care; clinical practice; clinically relevant; cytotoxicity; experience; genetic approach; in vivo; inpatient service; knock-down; medical schools; mortality; mouse model; nephrotoxicity; novel; novel strategies; preclinical study; prevent; public health relevance; success; therapeutic siRNA; therapeutic target; uptake

DESCRIPTION (provided by applicant): Acute inhibition of renal gene expression to prevent nephrotoxicity Prevention of AKI is of paramount importance, but success has been limited when approaches are applied generally. A key reason for these failures is a lack of targets specific to a given cause of AKI. This project utilizes a population genetics model to identify genes associated with sensitivity to four clinically relevant nephrotoxins: gentamicin, colistin, cisplatin, and clofarabine. In this proposal, we seek to validate the success of this population genetics model with acute in vivo siRNA inhibition of renal gene expression. The efficacy of target gene inhibition will be tested in corresponding murine models of drug-induced AKI to determine if this therapeutic strategy mitigates nephrotoxicity. A secondary aim of this project is to discern if acute inhibition of candidate transport genes similarly prevents nephrotoxicity by inhibiting proximal tubular uptake of gentamicin, colistin, cisplatin, or clofarabine. The expected outcome is to determine an optimal nephrotoxicity prevention strategy for each drug. The approach will be paradigm shifting - bringing a drug-specific intervention to each form of renal injury. The precedent for premedication to prevent nephrotoxicity is used in clinical practice as saline is often given to prevent contrast nephropathy. Thus, the siRNA strategies outlined in this proposal hold clinical relevance and the potential to translate to clinical trials. This approach wll yield multiple new molecular therapeutics specific to a variety of nephrotoxins, ready for rapid translation to clinical trials. My long-term goal is to become a physician scientist using large databases to uncover and validate genetic biomarkers and therapeutic targets which will readily translate to clinical care. Through this K08 award, I will learn new techniques including: analytic methods for genetic databases, animal model experimentation, laser micro-dissection (LMD), and in vivo siRNA knockdown. The acquired techniques will allow me to complete the above aims, but also provide the foundation to successfully compete for future R01 funding to: 1) uncover biomarkers from large genetic databases, 2) determine nephron segment specific gene expression with LMD of biopsy tissue, and 3) test mechanistic pathways and therapeutic targets in vivo with siRNA inhibition. These techniques can be applied to the improvement of renal care for many conditions. Further, the mentoring team I have assembled will advise me on the translation of these siRNA constructs to future clinical trials. My mentors and advisors at the Indiana University School of Medicine (IUSM) have a previous history of translating their pre-clinical studies with siRNA to successful phase 1 and 2 clinical trials. The rich environment at IUSM is sure to aid in my development as a physician scientist. With a dual appointment in the divisions of nephrology and clinical pharmacology, I am exposed to a wide range of collaborators in nearly every primary division. My division heads, Sharon Moe and David Flockhart, effectively protect my time as my clinical obligations are limited to 4 weeks of inpatient service and 1/2 day of clinic. They have provided ample laboratory space, start-up funds, and a technician. Our school renewed its NIH Clinical Translational Science Award (the Indiana CTSI) in 2013. I have already reaped the benefits of this advantage by obtaining a CTSI biomedical researcher grant. Finally, my mentors, Pierre Dagher and Todd Skaar, are incredibly supportive. The two of them provide a balance so I learn skills in both mouse models of acute kidney injury and pharmacogenomics analysis / validation. In summary, the proposed project builds logically on my fellowship training in nephrology and clinical pharmacology. My transition to Indiana University School of Medicine makes a great deal of sense not only because of its institutional commitment to me, but also what I can learn from my selected mentors, Drs. Pierre Dagher, Todd Skaar, and David Flockhart in order to address my specific aims and hypothesis. With ties to clinical pharmacology and nephrology, I am poised to apply novel strategies to nephrotoxicity prevention and other kidney diseases.

描述（由申请人提供）：急性抑制肾基因表达以预防肾毒性预防AKI至关重要，但当方法普遍应用时，成功有限。这些失败的一个关键原因是缺乏针对AKI特定原因的靶点。该项目利用群体遗传学模型来鉴定与对四种临床相关肾毒素敏感性相关的基因：庆大霉素、粘菌素、顺铂和氯法拉滨。在这个提议中，我们试图验证这个群体遗传学模型的成功与急性肾基因表达的体内siRNA抑制。将在药物诱导的AKI的相应鼠模型中测试靶基因抑制的功效，以确定该治疗策略是否减轻肾毒性。该项目的第二个目的是 辨别候选转运基因的急性抑制是否通过抑制庆大霉素、粘菌素、顺铂或氯法拉滨的近端肾小管摄取而类似地预防肾毒性。预期 结果是确定每种药物的最佳肾毒性预防策略。该方法将是范式转移-为每种形式的肾损伤带来药物特异性干预。在临床实践中使用预防肾毒性的前驱用药的先例，因为通常给予生理盐水以预防造影剂肾病。因此，本提案中概述的siRNA策略具有临床相关性，并有可能转化为临床试验。这种方法将产生多种新的分子治疗特异性的各种肾毒素，准备快速翻译到临床试验。我的长期目标是成为一名医生科学家，使用大型数据库来发现和验证遗传生物标志物和治疗靶点，这些标志物和靶点很容易转化为临床护理。通过这个K08奖，我将学习新的技术，包括：分析 用于遗传数据库、动物模型实验、激光显微切割（LMD）和体内siRNA敲除的方法。所获得的技术将使我能够完成上述目标，但也为成功竞争未来的R01资金提供了基础：1）从大型遗传数据库中发现生物标志物，2）用活检组织的LMD确定肾单位片段特异性基因表达，3）用siRNA抑制在体内测试机制途径和治疗靶点。这些技术可以应用于改善许多条件下的肾脏护理。此外，我组建的指导团队将为我提供关于将这些siRNA构建体转化为未来临床试验的建议。我在印第安纳州大学医学院（IUSM）的导师和顾问们有将siRNA临床前研究转化为成功的1期和2期临床试验的历史。在IUSM丰富的环境是一定要帮助我的发展作为一个医生科学家。由于在肾脏病学和临床药理学部门的双重任命，我接触到几乎每个主要部门的广泛合作者。我的部门负责人Sharon MoE和大卫弗洛克哈特有效地保护了我的时间，因为我的临床义务仅限于4周的住院服务和1/2天的门诊。他们提供了充足的实验室空间、启动资金和一名技术人员。我们学校在2013年更新了NIH临床转化科学奖（印第安纳州CTSI）。我已经通过获得CTSI生物医学研究人员补助金获得了这一优势的好处。最后，我的导师皮埃尔·达格尔和托德·斯卡尔非常支持我。他们两个提供了一个平衡，所以我学习技能，在这两个急性肾损伤和药物基因组学分析/验证小鼠模型。总而言之，我所提出的计画是建立在我在肾脏病学和临床药理学的实习训练基础上。我转到印第安纳州大学医学院是非常有意义的，不仅因为它对我的制度承诺，而且我可以从我选择的导师皮埃尔·达格尔博士、托德·斯卡尔博士和大卫·弗洛克哈特博士那里学到什么，以解决我的具体目标和假设。由于与临床药理学和肾脏病学的联系，我准备将新的策略应用于肾毒性预防和其他肾脏疾病。