Evaluation of a PPMO Inhibitor of Lassa Virus Infection and Disease in a Guinea Pig Model

在豚鼠模型中评价拉沙病毒感染和疾病的PPMO抑制剂 摘要： 拉沙热（Lassa fever，LF）是由拉沙病毒（Lassa virus，LASV）感染引起的一种严重的人类疾病 在西非大部分地区，每年造成数千人死亡。最近暴发的LF的特点是 与以前的疫情相比，病例数量增加，地理足迹扩大， 一场迅速发展的公共卫生危机此外，输入非流行国家的LF病例数为 此外，人们还担心LASV可能被用作生物战剂。目前还 没有许可的疫苗或特异性抗病毒治疗药物来干预LASV感染和治疗 LF和目前的治疗方案疗效不佳。因此，有一个迫切和紧急的医疗需要， 寻找新的抗病毒化合物来治疗LASV。肽缀合的磷酰二胺吗啉代寡聚物 （PPMO）是一类单链核酸类似物，其可以容易地进入细胞并干扰细胞内的蛋白质。 通过互补RNA的空间位阻产生病毒蛋白。PPMO已表现出相当大的 在体外和体内对许多RNA病毒，包括几种沙粒病毒的抗病毒功效。我们 我建议合成两个PPMO靶向LASV基因组末端的高度保守RNA序列 基因组将在非细胞毒性剂量范围内检测每种PPMO对几种遗传性疾病的抗病毒活性。 不同的LASV菌株在体外表现出最高抗病毒效力的PPMO将被推进到 在LASV疾病的豚鼠模型中进行测试。动物体重、体温、外观和行为，沿着 将监测几种组织中的死亡率和病毒滴度。本研究旨在评估 PPMO降低严重LASV疾病小动物模型的发病率和死亡率。如果成功， 该项目将产生一种抑制剂，可立即用作研究试剂，用于对抗任何菌株。 LASV，并作为进一步开发作为抗人类LASV感染的治疗剂的候选物。

Evaluation of a PPMO Inhibitor of Lassa Virus Infection and Disease in a Guinea Pig Model ABSTRACT: Lassa fever (LF) is caused by infection with Lassa virus (LASV), and is currently a major human disease problem in much of West Africa, causing thousands of deaths annually. Recent outbreaks of LF have been characterized by an increased number of cases and larger geographic footprint compared to previous outbreaks, and constitute a burgeoning public health crisis. In addition, the number of LF cases imported into non-endemic countries is growing, and there is also concern that LASV could be used as an agent of biological warfare. There is as yet no licensed vaccine or specific antiviral therapeutic to intervene against LASV-infections and treat patients with LF, and current treatment options have poor efficacy. There is therefore an immediate and critical medical need for new antiviral compounds to address LASV. Peptide-conjugated phosphorodiamidate morpholino oligomers (PPMO) are a class of single-stranded-nucleic acid analogs that can enter cells readily and interfere with the production of viral proteins through steric blocking of complementary RNA. PPMO have shown considerable antiviral efficacy in vitro and in vivo against a number of RNA viruses, including several arenaviruses. We propose to synthesize two PPMO targeting highly conserved RNA sequences in the genomic termini of the LASV genome. Each PPMO will be tested in a non-cytotoxic dose-range for antiviral activity against several genetically- disparate strains of LASV. The PPMO exhibiting the highest antiviral efficacy in vitro will then be advanced to testing in a guinea pig model of LASV disease. Animal weight, temperature, appearance and behavior, along with mortality and viral titer in several tissues will be monitored. This study is designed to evaluate the ability of PPMO to reduce morbidity and mortality in a small-animal model of of severe LASV-disease. If successful, the project will produce an inhibitor that would be immediately useful as a research reagent against any strain of LASV and as a candidate for further development as a therapeutic agent against LASV infections in humans.