Nanomicellar antiviral strategies for RSV infection

该计划的主要目标是进一步发展和转化生物学中的最新发现 呼吸道合胞病毒，一种感染约100万人的重要病原体 每年在全球范围内造成约20万人死亡。在美国，RSV导致11,000人死亡 每年都有老年人。目前还没有针对RSV感染的有效治疗方法或疫苗。目前，只有 高危婴儿接受以抗体为基础的预防，费用昂贵，效果适中 在减少住院方面。因此，一种广泛适用、有效和廉价的方法 预防或治疗呼吸道合胞病毒毛细支气管炎或肺炎仍然是一项尚未得到满足的迫切需要。 这项关于开发和测试一种新的预防和/或治疗RSV感染的建议是 灵感来自以下发现。I)磷脂胶束纳米颗粒(PMN)平台 被开发出来，当被鼻腔给药时，它主要将有效载荷输送到肺部。Ii)A 鉴定了一种能有效抑制呼吸道合胞病毒细胞的诱骗短七重复序列(HR)2肽 核聚变。Iii)人间充质细胞对呼吸道合胞病毒高度敏感。后者得到了帮助 在建立一种用于抗RSV药物筛选的新型3D支架中，它包括创建一个裸露的 小鼠肺支架(NMLS)脱细胞再细胞化的实验研究 想要的人类细胞，如hMSCs和上皮细胞，然后感染支架中的细胞 用呼吸道合胞病毒。IV)建立了一个健壮的免疫功能低下的小鼠模型 环磷酰胺治疗高粘液性RSV-L19F株感染。这些 研究进展导致假设RSV靶向PMN(RTPMN)结合HR2D 抗RSV-NS1和/或RSV-P基因的siRNAs，以及编码siRNAs的质粒可以 提供安全、有效和廉价的抗RSV预防和/或治疗。三个具体目标 美国国家航空航天局(SAS)将检验这些假设。 在SA#1中，多功能和智能的RTPMN将用编码的质粒合成 RSV-NS1和RSV-P基因核心和表面反融合HR2D多肽的siRNAs 并在体外进行表征。在SA#2中，将检查RTPMN的生物分布，pharm-tox 和PK/PD属性。SA#3将检查RTPMN的预防和治疗潜力- HR2D-psiNS1-P体外抗呼吸道合胞病毒感染及免疫抑制的实验研究 对小鼠的作用机制进行研究。 这些极具创新性的多学科翻译研究的结果有望 在人源化小鼠肺模型和模型中增加对RSV病理的理解 使用成年免疫功能低下的小鼠。抗病毒药物临床前配方研制工作圆满完成 基于RSV PMN的预防和治疗有望为IND驱动的研究铺平道路 和临床试验。

The major goal of this program is to further develop and translate recent discoveries in biology of respiratory syncytial virus (RSV), an important pathogen that infects an estimated 64 million people and causes ~200, 000 deaths globally every year. In the US, RSV causes >11,000 deaths of elderly annually. There is no effective treatment or vaccine against RSV infection. Currently, only high-risk infants receive antibody-based prophylaxis, which is expensive and moderately effective in reducing hospitalization. Therefore, a broadly applicable, effective and inexpensive approach to prevent or treat RSV-bronchiolitis or -pneumonia remains an urgent unmet need. This proposal to develop and test a novel prophylaxis and/or therapy against RSV infection was inspired by the following discoveries. i) A platform of phospholipid micellar nanoparticles (PMN) was developed, which when given intranasally delivers payload predominantly to the lung. ii) A decoy short heptad repeat (HR)2 peptide was identified, which effectively inhibits the RSV-cell fusion. iii) Human mesenchymal cells were found to be highly susceptible to RSV. The latter aided in establishing a novel 3D scaffold for anti-RSV drug screens, which consisted of creating a naked mouse lung scaffold (nMLS) by decellularization followed by recellularization of the nMLS with desired human cells, such as hMSCs and epithelial cells and then infecting the cells in scaffold with RSV. iv) A robust immunocompromised mouse model was created by combining cyclophosphamide treatment with infection by a highly mucogenic strain, RSV-L19F. These developments have led to the hypothesis that a RSV-targeted PMN (RTPMN), combining HR2D anti-fusion peptide, and plasmid encoded siRNAs against RSV-NS1 and/or RSV-P gene can provide a safe, effective and inexpensive anti-RSV prophylaxis and/or therapy. Three specific aims (SAs) will test these hypotheses. In SA#1, multifunctional and smart RTPMNs will be synthesized with plasmids encoding siRNAs for RSV-NS1and RSV-P genes in the core and anti-fusion HR2D peptides on the surface and be characterized in vitro. In SA#2, RTPMNs will be examined for their biodistribution, pharm-tox and PK/PD properties. SA#3 will examine the prophylactic and therapeutic potential of RTPMN- HR2D-psiNS1-P against RSV infection in an in vitro 3D lung scaffold and in immunocompromised mice to investigate the mechanism of action. The results of these highly innovative multidisciplinary translational studies are expected to increase the understanding of RSV pathology in humanized mouse lung model and in a model using adult immunocompromised mice. A successful completion of preclinical formulation of anti- RSV PMN-based prophylactics and therapeutics is expected to pave the way to IND-driven studies and clinical trials.