Influenza therapy by Au-nanorod 5'PPP-NS1-siRNA/cDNA targeting of bronchial cells

Au-nanorod 5PPP-NS1-siRNA/cDNA 靶向支气管细胞的流感治疗

基本信息

批准号：
7897618
负责人：
PAUL R KNIGHT III
金额：
$ 39.61万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-22 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7897618
关键词：
Abbreviations Acute Lung Injury Aerosols Alkaline Phosphatase Alveolar Alveolar Macrophages Animal Model Anti-Bacterial Agents Antibacterial Response Antiviral Agents Antiviral Response Avian Influenza Bacterial Infections Bacterial Pneumonia Bronchi Bronchoalveolar Lavage Cause of Death Cell Line Cells Cercopithecine Herpesvirus 1 Cessation of life Charge Chemistry Chronic Clinical Colony-forming units Comorbidity Complementary DNA Complex Congestive Heart Failure Disease Drug Formulations Drug resistance EMSA Electrophoretic Mobility Shift Assay Electrostatics Engineering Environment Epithelial Epithelial Cells Exhibits Fluorescence Gastrointestinal tract structure Gene Expression Gene Transfer Generations Genes Genetic Materials Glycolates Goals Gold Host Defense Human Immune Immune response Immunity Impairment In Vitro Individual Infection Inflammatory Inflammatory Response Influenza Influenza A Virus, H5N1 Subtype Inhalation Drug Administration Inhalators Injury Interferon Type I Interferons Intestines Investigational Drugs Kinetics Lead Lipopolysaccharides Lung Lung diseases Mediating Methods Modality Morbidity - disease rate Mouse Cell Line Mus Nanotechnology Natural Immunity Nonstructural Protein Nucleic Acids Pathway interactions Patients Peroxidases Phase Phase I Clinical Trials Play Production Quantum Dots Research Respiratory System Respiratory tract structure Risk Role Safety Secondary to Severities Small Interfering RNA Stress Surface Symptoms Therapeutic Tissues Toxic effect Trachea Translations Treatment Efficacy Viral Viral Genes Virulence Factors Virus Virus Replication Work anti-influenza base biomaterial compatibility body system bronchial epithelium cytokine design and construction flu improved in vitro activity in vivo influenza virulence influenza virus strain influenzavirus killings luminescence lung injury mortality mouse model nano nanoparticle nanorod nanoscale novel pandemic disease pathogen prevent promoter prophylactic public health relevance research study resistant strain respiratory response retinal rods treatment strategy tripolyphosphate two-photon type I interferon receptor

项目摘要

DESCRIPTION (provided by applicant): This application proposes to develop two novel prophylactic and therapeutic non-viral gene transfer strategies that target pulmonary cells in vivo employing nanotechnology. The lung is especially well suited for these treatment strategies as direct contact with the environment provides a portal for inhalation administration of cDNA and siRNA conjugated nanoplexes. The emergence of drug-resistant strains of human influenza A and B viruses, as well as avian H5N1 influenza viruses with pandemic potential to one or both classes of approved antiviral agents underscores the importance of developing novel antiviral strategies. The primary objective of the R21 phase is to construct an electrostatic complex between a cationic nanoparticle (i.e., Gold Nanorods, GNR) and anionic genetic material (i.e., cDNA or siRNA). These nanoplexes will be engineered such that they can be taken-up and express bioactivity in large airway (i.e., bronchial) epithelial cells with little or no untoward cellular or pulmonary responses. The siRNA/cDNA constructs, which have just recently been synthesized, have dual actions of suppressing the translation of the influenza virulence factor, NS1, as well as independently stimulating type I interferon production through activation of the RIG-I pathway. Stimulation of this antiviral innate immune pathway occurs as a result of a triphosphate (PPP) moiety attached to the 5' end of the siRNA. We will preferentially administer the GNR-5'PPP-NS1siRNA or its counterpart cDNA nanoplexes to the tracheal and bronchial epithelium in vivo, thereby increasing the safety of the treatment. Extension of these nanotechnological approaches can also be applied to treat other infectious, as well as non-infectious acute lung injuries. The focus in the R33 phase will be to demonstrate the therapeutic efficacy of using 5'PPP-NS1siRNA and cDNA-nanoplex targeting of large airway epithelial cells in vivo before and during influenza. In addition to assessing the clearance of influenza virus from the respiratory tract, the R33 phase will specifically examine the ability of 5'PPP-NS1siRNA or cDNA-nanoplexes to stimulate innate antiviral immunity, resulting in alteration of the inflammatory cytokine milieu, adaptive immune response, and antibacterial host defense, as well as prevent or reduce the degree of viral induced respiratory injury and impairment of bacterial clearance. We predict that these large airway epithelial-targeted nanoplexes will lead to prophylactic and therapeutic options that can prevent or significantly reduce the morbidity and severity of symptoms of influenza including the highly pathogenic H5N1 "bird flu" and the risk of secondary bacterial pneumonia, which is the major cause of death secondary to influenza. It is our goal to have a nanoparticle mediated novel antiviral prophylactic and therapeutic strategy at the completion of the R33 phase available for Investigational New Drug filing with the FDA to go for Phase 1 clinical trials as a result of the experiments proposed in this application. PUBLIC HEALTH RELEVANCE: Influenza is one of the top killers of people in the USA and the world, and the emergence of drug-resistant strains of influenza virus (including the "Bird Flu") requires that we develop new preventative and treatment approaches to this disease. This application proposes to develop a novel method to transfer a gene (cDNA) or its immediate message (siRNA) to cells that line the large airways of the lung employing an aerosol inhaler. Attaching cDNA or siRNA to small (nanometer size) gold rods will help deliver the anti-influenza treatment to inside the airway cells where it will work. This treatment stimulates immunity against influenza virus, as well as interferes with the virus' ability to do damage to the airways.

描述（由申请人提供）：本申请提出开发两种新颖的预防性和治疗性非病毒基因转移策略，利用纳米技术在体内靶向肺细胞。肺特别适合这些治疗策略，因为与环境的直接接触为吸入施用 cDNA 和 siRNA 缀合的纳米复合物提供了门户。人类甲型和乙型流感病毒耐药株的出现，以及对一类或两类已批准的抗病毒药物具有大流行潜力的禽 H5N1 流感病毒株的出现，强调了开发新型抗病毒策略的重要性。 R21 相的主要目标是在阳离子纳米颗粒（即金纳米棒、GNR）和阴离子遗传物质（即 cDNA 或 siRNA）之间构建静电复合物。这些纳米复合物将被设计成可以被大气道（即支气管）上皮细胞吸收并表达生物活性，而很少或没有不良的细胞或肺部反应。最近刚刚合成的 siRNA/cDNA 构建体具有抑制流感毒力因子 NS1 翻译以及通过激活 RIG-I 途径独立刺激 I 型干扰素产生的双重作用。这种抗病毒先天免疫途径的刺激是由于连接到 siRNA 5' 末端的三磷酸 (PPP) 部分而发生的。我们将优先将GNR-5'PPP-NS1siRNA或其对应的cDNA纳米复合物施用到体内气管和支气管上皮，从而提高治疗的安全性。这些纳米技术方法的扩展还可用于治疗其他感染性以及非感染性急性肺损伤。 R33 阶段的重点将是证明在流感之前和期间使用 5'PPP-NS1siRNA 和 cDNA-nanoplex 靶向大气道上皮细胞的体内治疗效果。除了评估流感病毒从呼吸道的清除情况外，R33 阶段还将专门检查 5'PPP-NS1siRNA 或 cDNA 纳米复合物刺激先天抗病毒免疫的能力，从而改变炎症细胞因子环境、适应性免疫反应和抗菌宿主防御，以及预防或减轻病毒引起的呼吸道损伤和细菌清除受损的程度。我们预测，这些针对大气道上皮的纳米复合物将带来预防和治疗选择，可以预防或显着降低流感症状的发病率和严重程度，包括高致病性 H5N1“禽流感”和继发性细菌性肺炎的风险。继发于流感的主要死亡原因。我们的目标是在 R33 阶段完成后，提供一种纳米颗粒介导的新型抗病毒预防和治疗策略，可用于向 FDA 提交研究性新药申请，以作为本申请中提出的实验的结果进行 1 期临床试验。公共卫生相关性：流感是美国和世界范围内人类的头号杀手之一，流感病毒耐药株（包括“禽流感”）的出现要求我们为此开发新的预防和治疗方法疾病。本申请提出开发一种新方法，使用气雾吸入器将基因 (cDNA) 或其即时信息 (siRNA) 转移至肺部大气道内的细胞。将 cDNA 或 siRNA 连接到小（纳米尺寸）金棒上将有助于将抗流感治疗药物传递到气道细胞内部，从而发挥作用。这种治疗可以刺激针对流感病毒的免疫力，并干扰病毒对呼吸道造成损害的能力。