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

• 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
• 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

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.

项目概要/摘要 急性肺损伤 (ALI) 及其更严重的急性呼吸窘迫综合征 (ARDS) 是一种 是重症患者呼吸衰竭的常见原因。目前所有针对 ALI/ARDS 的疗法均依赖于 支持性护理以改善临床结果。尚未开发出有效的药物。有一个迫切的需要 为 ALI/ARDS 开发安全、有效且基于更深入理解的新治疗策略 ALI 发病机制的相关研究。我们发现 MTOR 在 与 ALI 相关的炎症以及肺上皮细胞中 MTOR 的下调有可能 缓解这种炎症。然而，肺内皮细胞中 MTOR 的下调会产生相反的效果 并加剧炎症。因此，为了将这些发现转化为潜在的治疗方法，我们必须减少 肺上皮细胞中选择性的 MTOR 水平和活性。我们最近还报告了发现 二硫键约束的环状两亲肽 (CAPS) 与 siRNA 结合形成纳米复合物， 功能上影响 siRNA 货物向肺部的细胞内递送，以实现蛋白质沉默。我们假设 CAP-siRNA 纳米颗粒代表了通过以下方式将 siRNA 选择性递送至肺上皮细胞的理想载体： 简单的愿望。该提案的总体目标是表征细胞内 siRNA 的机制 通过 CAP-siRNA 纳米颗粒进行递送，并通过这些颗粒优化 MTOR 沉默，以验证 它们作为 ALI 药物治疗的应用。我们将利用跨学科的策略 完成既定的研究目标。该提案的具体目标是： 1) 描述 我们最近报道的 CAP 细胞内递送 siRNA 的易位机制。 2) 进行 结构活性研究，以优化 CAP 细胞内 siRNA 递送和基因沉默的效率 siRNA 纳米复合物。 3) 验证 CAP-siRNA 纳米粒子选择性敲低 MTOR 的用途 ALI 体内模型中的肺上皮细胞。拟议的工作需要物理方面的专业知识 和生物科学。研究团队利用肽设计、肺生物学和基因方面的专业知识 治疗。拟议的工作将扩展尼尔森、迪安和 拉赫曼团体。实现既定研究目标将解决在理解方面的重大差距 ALI的疾病病因学，验证MTOR下调治疗ALI的疗效，并提供 肽/siRNA 纳米粒子，有助于将 MTOR 特异性 siRNA 体内递送至肺部。此外， 提出的 CAP 药物代表了一类新型的创新细胞穿透肽基序，该基序简单且 生产成本低廉，并且不需要货物共价连接来促进细胞进入。这是 预计所提出的 CAP-siRNA 纳米颗粒也可用于其他肺部的基因沉默 作为平台技术，针对一系列疾病以及其他组织的目标。