Chlipoplex Nanoparticle Prophylactics for RSV Infection

描述（由申请人提供）：本提案的总体目标是开发一种对人类友好且更便宜的预防呼吸道合胞病毒（RSV）感染和其他呼吸道病毒的方法，该方法使用含pDNA的壳聚糖纳米颗粒经鼻给药，该方法允许ifn从头表达。在发现呼吸道合胞病毒45年后，目前还没有有效的疫苗或治疗方法可用于对抗呼吸道合胞病毒感染。目前RSV的预防包括每月使用RSV融合蛋白人源化抗体（Palivizumab, Synagis TM）进行治疗，该抗体仅对高危婴儿有中等效果，价格较高，并且其在成人和老年人中的应用尚不清楚。该建议的重点是预防的新概念，并基于发现鼻内给药编码ifn - γ (plfn - γ)的质粒DNA（pDNA）导致小鼠模型中RSV感染的显着减少。此外，外源性ifn - γ /可防止RSV感染人上皮细胞，并显著减少已建立的感染。此外，plfn - γ和plfn - β联合使用比单独使用更能显著降低上皮细胞的RSV感染。壳聚糖纳米颗粒可促进ifnγ基因在小鼠体内的转移。Transgenex Therapeutics （TGT）是一家初创的生物技术公司，正在开发一种叫做脂质体的生物聚合物，它可以保护pDNAs，并将它们靶向到病毒复制的上皮细胞、巨噬细胞或树突状细胞（dc）上。综上所述，这些发展导致了一种可行的假设，即脂质体将有效地递送pdna，而不会对鼻黏膜产生任何明显的不良影响，并提供有效的保护，防止病毒感染。这些假设将在本提案中根据以下具体目标进行检验。目标# 1。筛选和鉴定最有效的RSV感染plFN(s)。计划开发对人类友好的非整合pDNA载体，用于多种ifn，无论是单独的还是组合的，并在人类细胞系中测试它们的抗病毒特性。目标# 2。开发和测试脂质体以优化plfn的传递和表达。计划在模拟人类鼻黏膜的三维气液间期（ALl）培养物中开发含有pIFNs （CLIP）的脂质体，并测试其减毒RSV感染的有效性。目标# 3。在小鼠中测试所选的CLIP。我们计划在RSV感染小鼠模型中对选择的前两种clip进行测试，并研究它们在小鼠体内的抗病毒活性与剂量、安全性和免疫效应的关系。预计本文提出的研究结果将增强我们对CLIP作为预防RSV感染药物的潜力的理解，并将有希望的CLIP进入I期临床试验。

DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop a human-friendly and less-expensive method of prophylaxis for respiratory syncytial virus (RSV) infection and other respiratory viruses using intranasal administration of chitosan nanoparticles containing pDNA, which allows de novo expression of IFNs. Forty-five years after its discovery, no effective vaccine or treatment is currently available against RSV infection. Current prophylaxis for RSV comprises a monthly treatment with a humanized antibody to RSV fusion protein (Palivizumab, Synagis TM) that is only moderately effective in high risk infants, more expensive and its application to adults and elderly is unclear. This proposal focuses on a new concept for prophylaxis and is based on the discovery that intranasal administration of plasmid DNA(s) (pDNA) encoding IFN-gamma, (plFN-gamma) results in a significant decrease of RSV infection in a mouse model. Also, exogenous IFN-gamma/ prevents RSV infection of human epithelial cells and significantly decreases established infection. Also, a combination of plFN-gamma and plFN-beta induces more significant decrease in RSV infection of epithelial cells than either of them alone. Chitosan naoparticles have been developed that facilitate IFNgamma gene transfer in mice. The Transgenex Therapeutics (TGT), a start-up biotech company is developing biopolymers, called chlipoplexes, which protect the pDNAs and target them to the epithelial cells, macrophages or dendritic cells (DCs), where the virus replicates. Together, these developments have led to the working hypothesis that chlipoplexes will deliver effectively the pDNAs without any significant adverse effects in the nasal mucosa and provide significant protection against viral infections. These hypotheses will be tested in this proposal under the following specific aims. Aim #1. To screen and identify the most effective plFN(s) for RSV infection. It is planned to develop human-friendly, nonintegrating pDNA vectors for various IFNs, either singly or in combination and test their antiviral properties in human cell lines. Aim #2. To develop and test chlipoplexes for optimized delivery and expression of plFNs. It is planned to develop chlipoplexes containing pIFNs (CLIP) and test their effectiveness to attenuate RSV infection in the three dimensional air-liquid interphase (ALl) cultures, which mimics the nasal mucosa of humans. Aim #3. To test the selected CLIP(s) in mice. It is planned to test the selected top two CLIPs using the mouse model of RSV infection and investigate their antiviral activity in relation to dosage, safety, and immunological effects in mice. It is anticipated that the results of the studies proposed herein will enhance our understanding of the potential of CLIP as agents for prophylaxis against RSV infection and will move the promising CLIP(s) into phase I clinical trials.