Development of Nanotherapeutics for RSV-induced Lung Disease

RSV 引起的肺部疾病纳米疗法的开发

• 批准号: 8598008
• 负责人: Shyam S Mohapatra
• 金额: --
• 依托单位: JAMES A. HALEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-10-01 至 2015-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8598008
• 关键词: 3-Dimensional; A549; Address; Adult; Affect; Aging; Antiviral Agents; Apoptosis; Apoptotic; Area; Attenuated; Binding; Biology; Bronchiolitis; CCL4 gene; Caspase; Cell Culture Techniques; Cell Death; Cell Membrane Permeability; Cell Survival; Cells; Cessation of life; Chitosan; Chronic lung disease; Complex; Cyclophosphamide; Data; Development; Dissociation; Doctor of Philosophy; Effectiveness; Elderly; Encapsulated; Epithelial Cells; Fibroblasts; Fluorescein-5-isothiocyanate; Fluorescence Microscopy; Funding; Genes; Genetic; Goals; Grant Review; Host Defense; Hour; Human; Hybrids; Immune; Immune response; Immune system; Immunocompromised Host; In Vitro; Inbred BALB C Mice; Infant; Infection; Influenza; Intercellular adhesion molecule 1; Interferons; Knowledge; Label; Laboratories; Lead; Leadership; Link; Lipids; Lung; Lung Inflammation; Lung diseases; Membrane Potentials; Metabolic; Methods; Mitochondria; Modeling; Molecular Medicine; Motivation; Mucous Membrane; Mus; Natural Immunity; Nature; Nonstructural Protein; Palivizumab; Pathogenesis; Peptide Signal Sequences; Peptides; Physiological; Physiology; Plasmids; Play; Pneumonia; Postdoctoral Fellow; Principal Investigator; Process; Production; Prophylactic treatment; Proteins; Publications; RNA Helicase; RNA Interference; Recombinants; Regulation; Reporting; Research; Research Personnel; Research Project Grants; Respiratory Syncytial Virus Infections; Respiratory Syncytial Virus Vaccines; Respiratory syncytial virus; Role; Safety; Scientist; Signal Pathway; Signal Transduction; Signaling Protein; Small Interfering RNA; Structure-Activity Relationship; System; Technology; Testing; Therapeutic; Therapeutic Agents; Time; Tissue Model; Translations; Treatment Efficacy; Tretinoin; Veterans; Viral; Virus; Virus Diseases; Virus Replication; Work; accomplished suicide; attenuation; base; career; caspase-9; cytochrome c; effective therapy; efficacy testing; fetal; in vivo; mitochondrial membrane; mouse model; nanoparticle; nanotherapeutic; novel; novel therapeutics; pathogen; preclinical study; premature; prevent; professor; programs; promoter; protein structure function; receptor; red fluorescent protein; respiratory; response; skills; targeted delivery; vector control; viral RNA

DESCRIPTION (provided by applicant): The primary goal of this proposal is to investigate the mechanism whereby respiratory syncytial virus (RSV) nonstructural protein 1 (NS1) modulates innate immunity and cell survival through the mitochondrial system following RSV infection, and to combine this knowledge with nanoparticle technology to develop new ways to prevent and treat RSV infections. Veterans commonly have some form of chronic lung disease and are especially susceptible to respiratory viral infections like those caused by RSV and influenza. RSV is an opportunistic human respiratory pathogen that causes bronchiolitis in infants and pneumonia in immunocompromised adults and the elderly with an estimated 64 million infections and 166,000 deaths annually worldwide. RSV is becoming more and more of a serious threat to aging veterans. Despite progress, the precise nature of the RSV-induced innate immune response remains unclear. Our preliminary evidence shows that the NS1 protein associates with mitochondria and is found in complex with the mitochondrial antiviral signaling (MAVS) protein in RSV-infected A549 epithelial cells and inhibits the signaling of retinoic acid-inducible gene-I (RIG-I) and MAVS occurring via the caspase activation and recruitment domain (CARD). This interferes with IFN-2 production which inhibits the innate immune response. NS1 also blocks programmed cell death (apoptosis) of infected cells and prevents loss of mitochondrial membrane potential, thus promoting cell (and viral) survival. These data have led to the hypothesis that RSV utilizes NS1 to attenuate RIG-I- MAVS-induced antiviral IFN-2 production and to modulate mitochondrial function for increasing cell survival. The study of the precise mechanisms underlying these processes is expected to lead to the discovery of new targets for preventing or limiting RSV infection. To test these hypotheses, the following specific aims are proposed. In the first aim, it is planned to examine the role of NS1 in attenuating RIG-I/MAVS signaling during RSV infection. This includes analysis of the role of NS1 in regulating MAVS, LGP2 and RIG-1 expression and in attenuation of IFN-2 response during RSV infection, mitochondrial localization of NS1 protein and structure-function relationships of NS1- MAVS interaction. The second aim will focus on determining whether the NS1- MAVS interaction is required to prevent premature apoptosis and intrinsic cell death. This also includes addressing whether the NS1-MAVS complex is associated with anti- or pro-apoptotic factors and modulates their expression on the mitochondrial membrane. In the third aim, it is planned to develop and test targeted chlipid (chitosan-lipid) nanoparticles as a therapeutic agent for RSV-induced lung disease. These chlipids encapsulate a plasmid that encodes siRNAs for LGP2 and NS1 and a peptide for MAVS (referred to as pLMNS1), each of which individually has been shown to significantly down- regulate RSV replication in human cells. Lastly, in the fourth aim it is proposed to evaluate targeted nanoparticle-encapsulated pLMNS1 for treating RSV disease in a mouse model. The proposed research will be conducted by an excellent group of investigators with a proven track record in RSV disease, apoptosis and nanoparticle technology. The results are expected to lead to the discovery of novel targets and to the initiation of preclinical studies of these targets against RSV lung disease.

描述（由申请人提供）：