Immunotherapy for acute lung injury secondary to influenza

项目摘要/摘要 人类甲型和乙型流感病毒以及禽流感H5N1病毒耐药株的出现 具有大流行潜力的病毒，到唯一被批准的抗病毒药物，强调了开发 新的抗病毒策略。我们设计了阳离子纳米颗粒之间的静电络合物(即， 在IAV感染期间体内靶向呼吸道上皮细胞的阴离子RNA。这些纳米网络 在体内诱导针对IAV的抗病毒生物活性，对细胞或肺组织几乎或没有不良影响 回应。纳米复合体通过5‘-干扰素刺激早期的I型干扰素细胞反应。 三磷酸(PPP)-细胞内传感器RIG-I的RNA结合。此外，5‘PPP-NS1shRNA Nanopex配方抑制IAV毒力因子NS1的翻译，NS1抑制RIG-I和宿主 细胞RNA成熟。肺非常适合抗病毒纳米复合体策略，因为它提供了一个入口， 吸入给药的生物活性纳米网络。我们已经证明，这一策略在体内抑制IAV 复制治疗和避免“干扰素悖论”，特别是，减少IAV肺损伤和IAV 细菌从肺部清除的障碍。目前提案的重点是优化治疗方案 5‘PPP-NS1shRNA纳米复合体在体外和体内的作用。此外，我们建议携带 FDA一篇题为《抗病毒产品开发：进行和 向该机构提交病毒学研究报告。这包括体外和体内实验，以评估 治疗效果(抗病毒活性)、药代动力学、药物相互作用和病毒的发展 抵抗。此外，由于5‘PPP-NS1shRNA纳米复合体调节免疫反应，FDA 建议检查对免疫系统的作用可能导致的意外不良影响。 因此，我们还将识别被改变的特定免疫系统组件，以及评估 IAV感染的免疫介导的并发症包括呼吸道损伤的严重性增加， 以及与IAV相关的继发性细菌性肺炎的风险，这是流感病例死亡的主要原因。 具体地说，我们将检测5‘PPP-NS1shRNA纳米网络刺激先天抗病毒免疫的能力， 从而改变炎症细胞的渗透，炎性细胞因子环境，适应性免疫反应， 以及减少呼吸损伤和IAV相关的细菌清除障碍。除了……之外 评估IAV从呼吸道的清除情况，我们预测纳米网络结构将减少 具有抗药性的季节性和大流行性流感的发病率和症状严重程度 致病性H1N1猪源IAV病毒(S-OIV)和H5N1“禽流感”。这些纳米技术方法 还有可能治疗其他感染性(即埃博拉)或非感染性肺损伤。我们的提案是专门设计的 生产一种新的抗病毒纳米复合体制剂，用于第一阶段临床试验，作为研究性新药。

PROJECT SUMMARY/ABSTRACT The emergence of drug-resistant strains of human influenza A (IAV) and B viruses, as well as avian H5N1 virus with pandemic potential, to the only approved antiviral agents underscores the importance of developing novel antiviral strategies. We have engineered electrostatic complexes between cationic nanoparticles (i.e., chitosan) and anionic RNA that target airway epithelial cells in vivo during an IAV infection. These nanoplexes induce antiviral bioactivity directed against IAV in vivo with little or no untoward cellular or pulmonary responses. The nanoplex constructs stimulate early type I interferon (IFN) cellular responses through 5’- triphosphate (PPP)-RNA binding of the intracellular sensor, RIG-I. Additionally, the 5’PPP-NS1shRNA nanoplex formulation suppresses the translation of the IAV virulence factor, NS1, which inhibits RIG-I and host cell RNA maturation. The lung is well suited for an antiviral nanoplex strategy since it provides a portal for inhalation administration of bioactive nanoplexes. We have demonstrated that this strategy inhibits in vivo IAV replication therapeutically and avoids the “IFN paradox”, specifically, decreasing IAV lung injury, and IAV impairment of bacterial clearance from the lung. The focus of the current proposal is to optimize the therapeutic action of the 5’PPP-NS1shRNA nanoplex formulation in vitro and then in vivo. Additionally, we propose to carry out experiments recommended by the FDA article entitled “Antiviral Product Development: Conducting and Submitting Virological Studies to the Agency.” This includes in vitro and in vivo experiments to assess therapeutic efficacy (antiviral activity), pharmacokinetics, drug-drug interactions, and development of viral resistance. Additionally, because the 5’PPP-NS1shRNA nanoplex modulates the immune response, the FDA recommends examining possible unintended adverse effects resulting from actions on the immune system. Thus, we will also identify the specific immune system components that are altered, as well as assess the immune-mediated complications of an IAV infection including increased severity of the respiratory tract injury, and the risk of IAV-associated secondary bacterial pneumonia, a major cause of death in influenza cases. Specifically, we will examine the ability of 5’PPP-NS1shRNA nanoplexes to stimulate innate antiviral immunity, thereby changing infiltration of inflammatory cells, inflammatory cytokine milieu, adaptive immune responses, as well as decrease respiratory injury and IAV-associated impairment of bacterial clearance. In addition to assessing the clearance of IAV from the respiratory tract, we predict that the nanoplex construct will reduce the morbidity and severity of symptoms of influenza from drug resistant seasonal and pandemic strains, the highly pathogenic H1N1 swine-origin IAV virus (S-OIV) and H5N1 “bird flu”. These nano-technological approaches can also potentially treat other infectious (i.e., Ebola) or non-infectious lung injuries. Our proposal is designed to produce a novel antiviral nanoplex formulation for Phase 1 clinical trials as an Investigational New Drug.