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：
7688443
负责人：
PAUL R KNIGHT III
金额：
$ 38.54万
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/7688443
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 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）：本申请建议开发两种新型的预防性和治疗性的非病毒基因转移策略，这些基因转移策略靶向使用纳米技术的体内肺细胞。肺部特别适合这些治疗策略，因为与环境的直接接触为吸入cDNA和siRNA偶联纳米封装提供了门户。人类流感和B病毒的抗药性菌株的出现，以及对一类或两种认可的抗病毒药物具有大流行潜力的鸟类H5N1流感病毒的出现，强调了开发新型抗病毒药策略的重要性。 R21相的主要目标是在阳离子纳米颗粒（即金纳米棒，GNR）和阴离子遗传材料（即cDNA或siRNA）之间构造静电复合物。这些纳米旋转将进行设计，以使它们可以在大型气道（即支气管）上皮细胞中被吸收并表达生物活性，而细胞或没有弱的细胞或肺部反应。刚刚合成的siRNA/cDNA构建体具有抑制流感毒力因子NS1的翻译以及通过激活RIG-I途径的独立刺激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'端的三磷酸（PPP）部分而发生的。我们将优先将GNR-5'PPP-NS1SIRNA或其对应物cDNA纳米旋转到体内的气管和支气管上皮，从而提高了治疗的安全性。这些纳米技术方法的扩展也可以用于治疗其他感染性以及非感染性急性肺损伤。 R33阶段的重点是证明使用5'ppp-ns1siRNA和cDNA-nanoplex靶向在流感之前和期间对大气道上皮细胞的靶向的治疗功效。 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细菌清除的损害。我们预测，这些大型气道上皮靶向的纳米旋转将导致预防性和治疗方法，这些选择可以预防或显着降低流感症状的发病率和严重程度，包括高度致病性的H5N1“鸟流感”，以及次生细菌性肺炎的风险，这是死亡的主要导致二次造成了造成二次造成疫苗。我们的目标是在完成与FDA一起研究的R33期后，将纳米颗粒介导的新型抗病毒预防性和治疗策略，作为该应用程序中提出的实验的结果。公共卫生相关性：流感是美国和世界人民的最大杀手之一，并且出现了抗药性流感病毒菌株（包括“鸟流感”），要求我们开发新的预防和治疗方法来解决这种疾病。该应用建议开发一种新的方法，将基因（cDNA）或其直接信息（siRNA）传递到使用气溶胶吸入器的肺部大气道的细胞。将cDNA或siRNA连接到小的（纳米尺寸）金棒将有助于将抗激素治疗提供在气道细胞内的内部。这种治疗刺激了对流感病毒的免疫力，并干扰了该病毒对气道造成损害的能力。