Immunotherapy for acute lung injury secondary to influenza

项目概要/摘要 人类甲型流感病毒（IAV）和B型流感病毒以及禽流感H5 N1病毒抗药株的出现 病毒与流行的潜力，唯一批准的抗病毒药强调了发展的重要性， 新的抗病毒策略。我们已经设计了阳离子纳米颗粒之间的静电复合物（即， 壳聚糖）和在IAV感染期间体内靶向气道上皮细胞的阴离子RNA。这些纳米复合物 在体内诱导针对IAV抗病毒生物活性，而很少或没有不利的细胞或肺 应答所述纳米复合物构建体通过5 '-IFN-γ刺激早期I型干扰素（IFN）细胞应答。 细胞内传感器RIG-I的三磷酸（PPP）-RNA结合。此外，5 'PPP-NS 1 shRNA 纳米复合物制剂抑制IAV毒力因子NS 1的翻译，NS 1抑制RIG-I和宿主细胞的增殖。 细胞RNA成熟。肺非常适合抗病毒纳米复合物策略，因为它提供了一个门户， 生物活性纳米复合物的吸入给药。我们已经证明，这种策略抑制体内IAV 复制治疗和避免“干扰素悖论”，特别是，减少IAV肺损伤， 从肺部清除细菌的损害。目前建议的重点是优化治疗方案， 5 ′ PPP-NS 1 shRNA纳米复合物制剂在体外然后在体内的作用。此外，我们还建议 FDA文章“抗病毒产品开发：进行和 向机构提交病毒学研究报告。”这包括体外和体内实验， 治疗效果（抗病毒活性）、药代动力学、药物间相互作用和病毒的发展 阻力此外，由于5 'PPP-NS 1 shRNA纳米复合物调节免疫反应，FDA 建议检查对免疫系统的作用可能产生的意外不良影响。 因此，我们还将确定被改变的特定免疫系统成分，并评估免疫系统的功能。 IAV感染的免疫介导的并发症，包括呼吸道损伤的严重程度增加， 以及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。 具体来说，我们将研究5 'PPP-NS 1 shRNA纳米复合物刺激先天性抗病毒免疫的能力， 从而改变炎性细胞的浸润、炎性细胞因子环境、适应性免疫应答 以及减少呼吸道损伤和IAV相关的细菌清除障碍。除了 通过评估IAV从呼吸道的清除，我们预测纳米复合物构建体将减少IAV在呼吸道中的释放。 耐药季节性和大流行性流感病毒株的发病率和症状严重程度， 致病性H1N1猪源IAV病毒（S-OIV）和H5 N1“禽流感”。这些纳米技术方法 也可以潜在地治疗其它感染性（即，埃博拉病毒）或非感染性肺损伤。我们的建议旨在 生产一种新型的抗病毒纳米复合物制剂，作为研究性新药用于1期临床试验。

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.