Utilizing siRNA to Minimize Nephrotoxic Injury

利用 siRNA 最大限度地减少肾毒性损伤

• 批准号: 8107323
• 负责人: Simon J. Atkinson
• 金额: $ 38.5万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-11 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8107323
• 关键词: Acute Renal Failure with Renal Papillary Necrosis; Adverse effects; Affect; Albumins; Aminoglycosides; Animals; Attenuated; Biological; Catabolism; Cell Line; Cells; Chronic Kidney Failure; Cisplatin; Clinic; Clinical; Clinical Research; Development; Disease; Dynamin; End stage renal failure; Endocytosis; Epithelial Cells; Fluorescence Microscopy; Fluorescent Probes; Future; Gene Silencing; Image; Incidence; Injury; Kidney; Kidney Diseases; Kinetics; Label; Life; Light; Link; Liquid substance; Liver; Measures; Mediating; Methodology; Methods; Modeling; Multiple Myeloma; Nephrons; Nephrotoxic; Oligonucleotides; Organ; Pathway interactions; Phase; Physiology; Play; Prevention; Process; Proteins; Proximal Kidney Tubules; RNA; RNA Interference; Reagent; Renal corpuscle structure; Role; Secondary to; Small Interfering RNA; Technology; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Toxic effect; Toxin; Tracer; Tubular formation; cell injury; cell type; clinical application; clinically relevant; fluorophore; innovation; intravital microscopy; kidney cell; kidney epithelial cell; macromolecule; multi-photon; mutant; new technology; novel; novel therapeutics; prevent; protein expression; receptor; therapeutic gene; trafficking; translational approach; uptake

DESCRIPTION (provided by applicant): Nephrotoxins remain a major cause of acute kidney injury (AKI). RNA interference holds great promise as a novel and specific therapeutic gene-silencing technology for a wide range of diseases. Oligonucleotides, like many other small macromolecules, are present in the ultrafiltrate formed at the renal corpuscle, and therefore are presented to the tubular epithelial cells lining the nephron at relatively high concentrations. These cells, particularly those in the proximal tubule segments, avidly take up oligonucleotides, so that the kidney has by far the greatest accumulation of systemically delivered oligonucleotides. This makes siRNA a promising technology for treatment of kidney disease, especially given the particular sensitivity of proximal tubule cells to toxic or ischemic injury. Given the clinical and biological importance of these observations, the handling of siRNA by renal cell types needs to be better characterized. We propose that siRNA is preferentially accumulated in proximal tubule epithelial cells because of the specialization of these cells for rapid endocytosis of macromolecules delivered to the filtrate. New technologies in fluorescence microscopy, particularly the application of multi-photon imaging to intravital microscopy, allow the cellular and subcellular distribution of labeled oligonucleotides to be analyzed in real time in live animals. We propose to use this methodology in four specific aims to better understand the mechanism of uptake of siRNA by proximal tubule cells and to establish the feasibility of using siRNA to prevent toxic acute kidney injury (AKI). We propose: 1) to determine the kinetics of cellular and intracellular accumulation, catabolism and biological effects of siRNA in the normal kidney; 2) to determine the effect of chronic kidney disease on siRNA handling by the kidney; 3) to determine the effect of endocytic blockade on siRNA uptake; and 4) to test the ability of siRNA to prevent aminoglycoside uptake by proximal tubule cells and hence limit renal toxicity. PUBLIC HEALTH RELEVANCE: RNA interference is a method that can be used to stop the harmful effects of normal or mutant proteins, and may be an important and specific therapy for diseases in the future. The RNA molecules used in this technology are concentrated in the kidney, and the studies in this proposal will examine the mechanisms and consequences of this accumulation, the way that kidney disease can affect this process, and the possibility of using RNA to protect the kidney against toxins.

描述（由申请人提供）：肾毒素仍然是急性肾损伤（AKI）的主要原因。 RNA干扰是一种新型且特定的治疗基因沉淀技术，可用于多种疾病。与许多其他小的大分子一样，寡核苷酸也存在于肾球体形成的超滤中，因此将其呈现在相对较高浓度的肾小管上的肾小管上皮细胞中。这些细胞，尤其是近端小管片段中的细胞，狂热地占据寡核苷酸，因此肾脏已成为迄今为止全身递送的寡核苷酸的最大积累。这使得siRNA成为治疗肾脏疾病的有前途的技术，尤其是考虑到近端小管细胞对有毒或缺血性损伤的特殊敏感性。鉴于这些观察结果的临床和生物学重要性，需要更好地表征肾细胞类型处理siRNA。我们提出，由于这些细胞专门针对传递到滤液的大分子的快速内吞作用，因此优先积聚在近端小管上皮细胞中。荧光显微镜中的新技术，尤其是多光子成像在浸润中显微镜中的应用，可以在活动物中实时分析标记的寡核苷酸的细胞和亚细胞分布。我们建议在四个特定目的中使用这种方法，以更好地了解近端小管细胞吸收siRNA的机制，并确定使用siRNA预防有毒急性肾脏损伤（AKI）的可行性。我们建议：1）确定siRNA在正常肾脏中细胞和细胞内积累，分解代谢和生物学作用的动力学； 2）确定慢性肾脏疾病对肾脏处理的影响； 3）确定内吞封锁对siRNA摄取的影响； 4）测试siRNA预防近端小管细胞摄取氨基糖苷的能力，从而限制肾脏毒性。 公共卫生相关性：RNA干扰是一种可用于阻止正常蛋白或突变蛋白的有害作用的方法，并且可能是对疾病的重要疗法。该技术中使用的RNA分子集中在肾脏中，该提案中的研究将检查这种积累的机制和后果，肾脏疾病会影响这一过程的方式以及使用RNA保护肾脏免受毒素的可能性。