Utilizing siRNA to Minimize Nephrotoxic Injury

利用 siRNA 最大限度地减少肾毒性损伤

• 批准号: 8675226
• 负责人: Simon J. Atkinson
• 金额: $ 33.5万
• 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 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8675226
• 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.

描述（由申请方提供）：肾毒素仍然是急性肾损伤（阿基）的主要原因。RNA干扰作为一种新的和特异性的治疗基因沉默技术，对广泛的疾病有很大的希望。寡核苷酸，像许多其他小的大分子一样，存在于肾小体形成的超滤液中，因此以相对高的浓度呈递给肾单位衬里的肾小管上皮细胞。这些细胞，特别是近端小管节段中的那些细胞，贪婪地摄取寡核苷酸，使得肾脏迄今为止具有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景的技术，特别是考虑到近端小管细胞对毒性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对siRNA的处理。我们提出，siRNA是优先积累在近端小管上皮细胞，因为这些细胞的专业化快速内吞的大分子传递到滤液。荧光显微镜的新技术，特别是多光子成像活体显微镜的应用，允许标记的寡核苷酸的细胞和亚细胞分布进行分析，在活的动物在真实的时间。我们建议在四个具体目标中使用这种方法，以更好地了解近端小管细胞摄取siRNA的机制，并建立使用siRNA预防毒性急性肾损伤（阿基）的可行性。我们建议：1）确定siRNA在正常肾脏中的细胞和细胞内积累的动力学、催化作用和生物学效应; 2）确定慢性肾病对肾脏处理siRNA的影响; 3）确定内吞阻断对siRNA摄取的影响;和4）测试siRNA阻止近端小管细胞摄取氨基糖苷并因此限制肾毒性的能力。