Cell-specific Delivery of RNAi to Pulmonary Alveolar Macrophages in vivo

RNAi 细胞特异性递送至体内肺泡巨噬细胞

• 批准号: 7295731
• 负责人: Darrell N. Kotton
• 金额: $ 19.72万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7295731
• 关键词: Actins; Adverse effects; Alveolar; Alveolar Macrophages; Anti-Inflammatory Agents; Anti-inflammatory; Apoptosis; Applications Grants; Asthma; B-Lymphocytes; Bronchoalveolar Lavage; CCL2 gene; CXCL11 gene; Cell Lineage; Cell Separation; Cells; Data; Digestion; Disease; Endothelial Cells; Endothelium; Epithelial; Epithelium; Exposure to; FGF2 gene; Fibroblast Growth Factor 2; Flow Cytometry; Fluorescence Microscopy; Frozen Sections; Gene Expression; Genes; Green Fluorescent Proteins; Immune; In Vitro; Infection; Inflammation; Inflammatory; Influenza; Interferons; Interleukin-10; Interleukin-6; Lung; Lung Inflammation; Lung diseases; MHC Class II Genes; Mammalian Cell; Measures; Messenger RNA; Methods; Mus; Myelogenous; NF-kappa B; Numbers; Organ; Parainfluenza; Pathway interactions; Pneumonia; Population; Proteins; Pulmonary Emphysema; RNA Interference; RNA Interference Pathway; Reporter; Reporter Genes; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Rodent; Route; Screening procedure; Signal Transduction; Small Interfering RNA; Smoke; Specificity; Structure of parenchyma of lung; Subfamily lentivirinae; Surface; System; Systems Biology; T-Lymphocyte; Technology; Testing; Tetracycline; Tetracyclines; Time; Tissues; Tobacco; Tobacco smoke; Transgenic Mice; Viral; base; cell type; cytokine; design; in vivo; novel; p65; pol genes; programs; research study; small hairpin RNA; targeted delivery

DESCRIPTION (provided by applicant): The delivery of interfering RNAs to lung tissue provides an opportunity to develop novel anti-viral and anti-inflammatory therapies for a variety of lung disorders. Intratracheal delivery of constructs for RNA interference (RNAi), as has been used to block influenza and parainfluenza lung infections, is a particularly exciting approach to treating lung disease because it offers potentially tissue-specific delivery of these molecules to a target organ. However, the cell types transduced with these methods are not yet known, and some investigators have found concerning non-specific effects in non-pulmonary tissues of mice exposed to this route of delivery. In addition, the potential for adverse effects in the lung, such as activation of inflammation are concerns that limit application of RNAi technology. Cell-specific, tissue-specific delivery of RNAi to immune cells within the lung would be a significant advance enabling potential treatment of many diseases characterized by inflammation, including asthma and emphysema. This grant application presents preliminary data demonstrating a novel lentiviral-based system that allows the cell-specific, tissue-specific in vivo delivery of RNAi to resident alveolar macrophages. The specific aims of this proposal are designed to test the efficacy of this targeted in vivo delivery method and to understand the biology of this system, including screening for specificity and potential off-target effects. Finally, the system is applied for in vivo knockdown of NF-kB signaling in mouse alveolar macrophages and the treatment of tobacco smoke- induced lung inflammation.

描述(由申请人提供)：将干扰RNA输送到肺组织为开发针对各种肺部疾病的新型抗病毒和抗炎疗法提供了机会。用于阻断流感和副流感肺部感染的RNA干扰构建体(RNAi)经气管内递送是治疗肺部疾病的一种特别令人兴奋的方法，因为它提供了潜在的组织特异性递送这些分子到靶器官。然而，通过这些方法转导的细胞类型尚不清楚，一些研究人员发现，暴露于这种传递途径的小鼠的非肺组织中存在非特异性影响。此外，肺部潜在的不良反应，如炎症的激活，是限制RNAi技术应用的担忧。细胞特异性、组织特异性的RNAi传递到肺内的免疫细胞将是一项重大进步，能够潜在地治疗许多以炎症为特征的疾病，包括哮喘和肺气肿。这项资助申请展示了一种新的基于慢病毒的系统，该系统允许在体内将RNAi特定于细胞、组织特定的RNAi传递到常驻的肺泡巨噬细胞。这项建议的具体目的是测试这种靶向体内给药方法的有效性，并了解该系统的生物学，包括筛选特异性和潜在的靶外效应。最后，该系统被用于体内敲除小鼠肺泡巨噬细胞中的核因子-kB信号，并用于治疗烟草烟雾引起的肺部炎症。