Nanomicellar antiviral strategies for RSV infection

该计划的主要目标是进一步发展和翻译生物学的最新发现， 呼吸道合胞病毒（RSV）是一种重要的病原体，估计感染了6400万人 全球每年约有20万人因此死亡。在美国，RSV导致> 11，000例死亡， 老年人每年目前还没有针对RSV感染的有效治疗方法或疫苗。目前只有 高危婴儿接受基于抗体的预防治疗，这种治疗费用昂贵，但效果一般 减少住院治疗。因此，一个广泛适用、有效和廉价的方法， 预防或治疗RSV细支气管炎或肺炎仍然是一个迫切的未满足的需求。 该开发和测试针对RSV感染的新型预防和/或治疗的提议是 受到以下发现的启发。i）磷脂胶束纳米颗粒（PMN）平台 开发了一种鼻内给药时主要将有效载荷递送至肺部的药物。（二）A 诱饵短七肽重复序列（HR）2肽被鉴定，其有效地抑制RSV-细胞 核聚变iii）发现人类间充质细胞对RSV高度易感。后者帮助 在建立用于抗RSV药物筛选的新型3D支架中， 小鼠肺支架（nMLS），通过去细胞化，然后用 所需的人细胞，如hMSC和上皮细胞，然后感染支架中的细胞， 与RSV iv）通过组合以下物质来创建稳健的免疫受损小鼠模型： 环磷酰胺治疗与高粘液生成菌株RSV-L19 F感染。这些 发展导致了一种假设，即RSV靶向PMN（RTPMN），结合HR 2D 抗融合肽和质粒编码的抗RSV-NS 1和/或RSV-P基因的siRNA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 (SAs)将检验这些假设。 在SA#1中，多功能和智能RTPMN将用编码 核心中的RSV-NS 1和RSV-P基因的siRNA和表面上的抗融合HR 2D肽 并在体外进行表征。在SA#2中，将检查RTPMN的生物分布、药物毒性 和PK/PD特性。SA#3将检查RTPMN的预防和治疗潜力- HR 2D-psiNS 1-P在体外3D肺支架和免疫功能低下的细胞中对抗RSV感染 小鼠研究其作用机制。 这些高度创新的多学科翻译研究的结果预计将 增加对人源化小鼠肺模型和模型中RSV病理学的理解 使用成年免疫缺陷小鼠。成功完成抗- 基于RSV PMN的药物和治疗有望为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.