Novel Peptide/siRNA Nanoparticles for Treatment of Acute Lung Injury

用于治疗急性肺损伤的新型肽/siRNA纳米颗粒

基本信息

批准号：
9376455
负责人：
David A Dean
金额：
$ 59.24万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-07-01 至 2021-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9376455
关键词：
Acute Acute Lung Injury Address Adult Respiratory Distress Syndrome Aerosols Affect Affinity Air Alveolar Cell Animal Model Binding Biochemical Genetics Biological Biological Sciences Biology Blood Vessels Caliber Case Study Cells Cellular biology Charge Chemicals Chemistry Clinical Complex Critical Illness Data Development Disease Disulfides Down-Regulation Endothelial Cells Endothelium Epithelial Epithelial Cells Epithelium Etiology Explosion FRAP1 gene Functional disorder Gene Silencing Gene Transfer Goals Hydrophobicity In Vitro Inflammation Inflammatory Injury Intervention Intracellular translocation Length Length of Stay Lung Lung Inflammation Mission Modality Modeling Mus Outcome Pathogenesis Patients Peptides Periodicity Pharmaceutical Preparations Pharmacological Treatment Pharmacology Play Proteins Public Health Pulmonary Edema Pulmonary Gas Exchange RNA Interference Reagent Reporting Research Resolution Respiratory Failure Role Small Interfering RNA Structure Supportive care Technology Testing Therapeutic Tissues Translating Validation Work base cell injury cell type design effective therapy gene therapy genetic approach improved in vivo in vivo Model innovation knock-down lung injury mortality nanoparticle novel overexpression particle physical science small hairpin RNA treatment strategy vector

Acute Acute Lung Injury Address Adult Respiratory Distress Syndrome Aerosols Affect Affinity Air Alveolar Cell Animal Model Binding Biochemical Genetics Biological Biological Sciences Biology Blood Vessels Caliber Case Study Cells Cellular biology Charge Chemicals Chemistry Clinical Complex Critical Illness Data Development Disease Disulfides Down-Regulation Endothelial Cells Endothelium Epithelial Epithelial Cells Epithelium Etiology Explosion FRAP1 gene Functional disorder Gene Silencing Gene Transfer Goals Hydrophobicity In Vitro Inflammation Inflammatory Injury Intervention Intracellular translocation Length Length of Stay Lung Lung Inflammation Mission Modality Modeling Mus Outcome Pathogenesis Patients Peptides Periodicity Pharmaceutical Preparations Pharmacological Treatment Pharmacology Play Proteins Public Health Pulmonary Edema Pulmonary Gas Exchange RNA Interference Reagent Reporting Research Resolution Respiratory Failure Role Small Interfering RNA Structure Supportive care Technology Testing Therapeutic Tissues Translating Validation Work base cell injury cell type design effective therapy gene therapy genetic approach improved in vivo in vivo Model innovation knock-down lung injury mortality nanoparticle novel overexpression particle physical science small hairpin RNA treatment strategy vector

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项目摘要

Project Summary/Abstract Acute Lung Injury (ALI) and its more severe form Acute Respiratory Distress Syndrome (ARDS) are a common cause of respiratory failure in critically ill patients. All current therapies for ALI/ARDS rely on supportive care to improve clinical outcome. No effective drugs have been developed. There is an urgent need to develop new treatment strategies for ALI/ARDS that are safe, effective, and based on deeper understanding of the mechanisms involved in ALI pathogenesis. We have discovered that MTOR plays a key role in the inflammation associated with ALI and that downregulation of MTOR in lung epithelial cells has the potential to alleviate this inflammation. However, downregulation of MTOR in lung endothelial cells has the opposite effect and exacerbates inflammation. Thus, to translate these findings into a potential treatment, we must reduce MTOR levels and activity selectively in the lung epithelium. We have also recently reported the discovery of disulfide-constrained, cyclic amphipathic peptides (CAPS) that bind to siRNA to form nanocomplexes that can functionally affect intracellular delivery of siRNA cargo to the lung for protein silencing. We hypothesize that CAP-siRNA nanoparticles represent an ideal vector for selective delivery of siRNA to lung epithelial cells by simple aspiration. The overall objective of this proposal is to characterize the mechanism of intracellular siRNA delivery by CAP-siRNA nanoparticles and to optimize MTOR silencing by these particles toward validation of their application as a pharmacologic treatment for ALI. We will utilize a cross-disciplinary strategy to accomplish the stated research objective. The Specific Aims of the proposal are: 1) To characterize the mechanism of translocation for intracellular delivery of siRNA by our recently reported CAPs. 2) To conduct structure-activity studies to optimize the efficiency of intracellular siRNA delivery and gene silencing by CAP- siRNA nanocomplexes. 3) To validate the use of CAP-siRNA nanoparticles for selective knockdown of MTOR in lung epithelial cells in an in vivo model for ALI. The proposed work requires expertise in both the physical and biological sciences. The research team draws on expertise in peptide design, lung biology, and gene therapy. The proposed work will extend existing collaborative relationships between the Nilsson, Dean, and Rahman groups. Accomplishment of the stated research goal will address significant gaps in understanding of the disease etiology of ALI, validate the efficacy of MTOR downregulation for treatment of ALI, and provide peptide/siRNA nanoparticles that facilitate in vivo delivery of MTOR-specific siRNA to the lung. Further, the proposed CAP agents represent a new class of innovative cell-penetrating peptide motif that is simple and inexpensive to produce and that does not require covalent attachment of cargo to promote cell entry. It is anticipated that the proposed CAP-siRNA nanoparticles will be also useful for gene silencing of other lung targets in a range of disorders as well as in other tissues as a platform technology.

项目摘要/摘要急性肺损伤（ALI）及其更严重的急性呼吸窘迫综合征（ARDS）是重症患者呼吸衰竭的常见原因。所有目前针对ALI/ARDS的疗法依赖支持护理以改善临床结果。没有开发出有效的药物。迫切需要为安全，有效且基于更深入的理解的ALI/ARD制定新的治疗策略 ALI发病机理所涉及的机制。我们发现MTOR在与Ali相关的炎症以及肺上皮细胞中MTOR的下调具有潜力减轻这种炎症。但是，肺内皮细胞中MTOR的下调具有相反的作用并加剧炎症。因此，要将这些发现转化为潜在的治疗方法，我们必须减少 MTOR水平和活性在肺上皮中有选择性。我们最近还报道了发现二硫键约束的环状两亲性肽（CAP）与siRNA结合以形成可以形成的纳米复合体在功能上影响siRNA货物向肺部的细胞内输送，以进行蛋白质沉默。我们假设这一点 Cap-SiRNA纳米颗粒代表了选择性递送siRNA向肺上皮细胞的理想载体简单的抽吸。该提议的总体目的是表征细胞内siRNA的机制通过Cap-siRNA纳米颗粒传递，并通过这些颗粒优化MTOR沉默他们作为ALI的药理治疗的应用。我们将利用跨学科策略来完成既定的研究目标。该提案的具体目的是：1）表征我们最近报道的CAPS的细胞内递送siRNA的易位机制。 2）进行结构活性研究以优化细胞内siRNA递送和基因沉默的效率 siRNA纳米复合体。 3）验证使用Cap-SiRNA纳米颗粒以选择性敲低mTOR 在ALI体内模型中的肺上皮细胞中。拟议的工作需要在物理上的专业知识和生物科学。研究小组借鉴了肽设计，肺部生物学和基因的专业知识治疗。拟议的工作将扩展尼尔森，院长和拉赫曼团体。既定研究目标的实现将解决理解的重大差距 ALI疾病病因验证MTOR下调对Ali治疗的功效，并提供肽/siRNA纳米颗粒，可促进体内MTOR特异性siRNA向肺部的递送。此外，提出的瓶盖剂代表了一种新的创新细胞穿透肽基序，很简单且廉价生产，这不需要共价附着货物来促进细胞进入。这是预计所提出的cap-siRNA纳米颗粒也将用于其他肺的基因沉默在各种疾病以及其他组织中的目标是一种平台技术。