Ultrasound-triggered delivery of siRNA as treatment for diabetic kidney disease

超声波触发 siRNA 治疗糖尿病肾病

• 批准号: 8201249
• 负责人: Joshua J. Rychak
• 金额: $ 78.49万
• 依托单位: TARGESON, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8201249
• 关键词: Acoustics; Acute; Aftercare; Animal Disease Models; Animal Model; Benchmarking; Binding; Biodistribution; Biological Assay; Blinded; Blocking Antibodies; Blood; Cell Death; Cells; Chronic; Clinical; Collagen; Data; Development; Diabetic Nephropathy; Diabetic mouse; Disease; Dose; Drug Formulations; Electroporation; Employee Strikes; Endothelial Cells; Enzyme-Linked Immunosorbent Assay; Evaluation; Exhibits; Exposure to; Fibrinogen; Fibrosis; Flow Cytometry; Fluorescence; Growth Factor; Hour; Human; Image; Immunohistochemistry; In Situ Hybridization; In Vitro; Incubated; Industry; Insulin-Dependent Diabetes Mellitus; Kidney; Kidney Diseases; Label; Ligands; Liver; Lung; Measures; Mediating; Methods; Microbubbles; Modeling; Molecular Target; Morphology; Mus; Non-Insulin-Dependent Diabetes Mellitus; Nucleic Acids; Pathologist; Perfusion; Plasma; Plasmids; Play; Process; Production; Proteins; Proteinuria; Residual state; Resistance; Ribonucleases; Rodent; Role; Schedule; Selectins; Small Interfering RNA; Spleen; Staging; Staining method; Stains; Surface; Techniques; Technology; Therapeutic; Time; Tissues; Toxic effect; Transforming Growth Factor beta; Treatment Protocols; Ultrasonic Therapy; Ultrasonography; Validation; Work; base; cell type; clinically relevant; cytotoxicity; diabetic; gel electrophoresis; gene therapy; head-to-head comparison; in vivo; kidney vascular structure; knock-down; mesangial cell; neutralizing antibody; non-invasive monitor; novel; nuclease; research study; small hairpin RNA; sonoporation; success; targeted delivery; terthienyl; zeta potential

DESCRIPTION (provided by applicant): Gene therapy in the context of diabetic kidney disease holds tremendous promise, although concrete progress in the field has been difficult to achieve. Specifically, the growth factor TGF-2 is known to play a critical role in glomerular matrix expansion and progression of glomerular fibrosis, and inhibition of TGF-2 activity by neutralizing antibodies has shown striking efficacy in several animal models of diabetic kidney disease. However, as is the case with many targets of molecular therapy, TGF-2 is strongly implicated in numerous somatic processes, and off-target inhibition may pose severe detrimental effects. Means by which TGF-2 therapy could be targeted specifically to the impaired kidney could offer a critical breakthrough in making gene therapy for kidney disease a reality. In the current proposal, we aim to develop a novel microbubble-based delivery vehicle for targeted TGF-2 gene therapy. We have developed a microbubble that can be conjugated with a payload of plasmid or siRNA, and targeted to the diabetic kidney by means of a selectin-binding targeting ligand. These agents can be administered intravenously, and their accumulation within the kidney monitored by non-invasive ultrasound imaging. Release of the siRNA payload and transfer into the targeted glomerular cells is mediated by application of high-power ultrasound energy specifically to the kidneys, which causes rapid destruction of the microbubbles and a transient poration of the adjacent cells. Remaining untargeted agents are cleared to the liver, spleen, and lung, where agent destruction by deflation exposes any residual siRNA to endogenous nucleases, thus potentially reducing off-target effects. We aim to evaluate this targeted siRNA delivery strategy in several clinically-relevant mouse models of diabetes. TGF-2 knock-down, and reduction in mesangial matrix expansion, will be assessed longitudinally. We will also systematically assess the payload capacity and stability of the agent, and evaluate toxicity and biodistribution in rodents. We anticipate that successful completion of the proposed aims will demonstrate efficacy of this targeted delivery technology for treatment of diabetic kidney disease, and provide Targeson with critical data needed to advance to the next stage of development for clinical use. PUBLIC HEALTH RELEVANCE: Few techniques for targeted gene therapy are available, especially in deep tissues such as the kidney. This project will examine the ability of a targeted ultrasound-based technique to deliver a therapeutic siRNA to diabetic kidney, and determine whether this strategy is able to ameliorate glomerular fibrosis in animal models of the disease.

描述（由申请人提供）：尽管该领域很难取得具体进展，但糖尿病肾病的基因治疗具有巨大的前景。具体而言，生长因子 TGF-2 已知在肾小球基质扩张和肾小球纤维化进展中发挥关键作用，并且通过中和抗体抑制 TGF-2 活性在几种糖尿病肾病动物模型中显示出惊人的功效。然而，与分子治疗的许多靶点一样，TGF-2 与许多躯体过程密切相关，脱靶抑制可能会造成严重的有害影响。 TGF-2疗法可以专门针对受损肾脏进行治疗，这可能为肾病基因治疗成为现实提供关键突破。在当前的提案中，我们的目标是开发一种新型的基于微泡的递送载体，用于靶向 TGF-2 基因治疗。我们开发了一种微泡，可以与质粒或 siRNA 的有效负载缀合，并通过选择素结合靶向配体靶向糖尿病肾脏。这些药物可以静脉注射，并通过非侵入性超声成像监测它们在肾脏内的积累。 siRNA 有效负载的释放并转移到目标肾小球细胞中是通过将高功率超声能量专门应用于肾脏来介导的，这会导致微泡的快速破坏和邻近细胞的瞬时穿孔。剩余的非靶向药物被清除到肝脏、脾脏和肺，其中通气破坏药物会使任何残留的 siRNA 暴露于内源性核酸酶，从而潜在地减少脱靶效应。我们的目标是在几种临床相关的糖尿病小鼠模型中评估这种靶向 siRNA 递送策略。 TGF-2 敲低和系膜基质扩张减少将进行纵向评估。我们还将系统地评估该药剂的有效负载能力和稳定性，并评估其在啮齿动物中的毒性和生物分布。我们预计，成功完成拟议目标将证明这种靶向递送技术治疗糖尿病肾病的功效，并为 Targeson 提供进入下一阶段临床应用开发所需的关键数据。 公共卫生相关性：目前可用的靶向基因治疗技术很少，特别是在肾脏等深层组织中。该项目将检查基于超声的靶向技术向糖尿病肾脏递送治疗性 siRNA 的能力，并确定该策略是否能够改善该疾病动物模型中的肾小球纤维化。