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

• 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
• 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

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的预防性和治疗性非病毒基因转移策略，其采用纳米技术在体内靶向肺细胞。肺特别适合于这些治疗策略，因为与环境的直接接触提供了吸入施用cDNA和siRNA缀合的纳米复合物的入口。人类甲型和B型流感病毒以及禽H5 N1流感病毒的耐药株的出现，对一类或两类已批准的抗病毒药物具有大流行潜力，这突出了开发新型抗病毒策略的重要性。R21相的主要目的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）。这些纳米复合物将被工程化，使得它们可以被摄取并在大气道中表达生物活性（即，支气管）上皮细胞，很少或没有不良的细胞或肺反应。最近合成的siRNA/cDNA构建体具有抑制流感毒力因子NS 1的翻译以及通过激活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'端的三磷酸（PPP）部分而发生的。我们将优先将GNR-5 ′ PPP-NS 1 siRNA或其对应物cDNA纳米复合物施用至体内气管和支气管上皮，从而增加治疗的安全性。这些纳米技术方法的扩展也可以应用于治疗其他感染性和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-NS 1 siRNA和cDNA-nanoplex靶向体内大气道上皮细胞的治疗功效。除了评估流感病毒从呼吸道的清除外，R33阶段还将专门检查5 'PPP-NS 1 siRNA或cDNA-纳米复合物刺激先天性抗病毒免疫的能力，从而导致炎性细胞因子环境、适应性免疫应答和抗菌宿主防御的改变，以及预防或降低病毒诱导的呼吸道损伤和细菌清除受损的程度。我们预测，这些大气道上皮靶向纳米复合物将导致预防和治疗选择，可以预防或显著降低流感症状的发病率和严重程度，包括高致病性H5 N1“禽流感”和继发性细菌性肺炎的风险，这是继发于流感的主要死亡原因。我们的目标是在R33阶段完成时获得纳米颗粒介导的新型抗病毒预防和治疗策略，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：流感是美国和世界上人类的头号杀手之一，流感病毒（包括“禽流感”）的耐药株的出现要求我们开发新的预防和治疗方法来治疗这种疾病。本申请提出开发一种新的方法，使用气雾剂吸入器将基因（cDNA）或其直接信息（siRNA）转移到排列在肺的大气道上的细胞。将cDNA或siRNA连接到小的（纳米尺寸）金棒上将有助于将抗流感治疗传递到气道细胞内部，在那里它将起作用。这种治疗刺激了对流感病毒的免疫力，并干扰了病毒对呼吸道造成损害的能力。