Development of Gene-Silencing Therapeutics for Pseudomonas aeruginosa

项目摘要/摘要 革兰氏阴性病原体对许多抗菌药的抗药性越来越强。此外，这条管道 因为新的抗生素规模很小，迫切需要新的治疗方法。这在病人身上可能尤其成问题。 患有慢性感染或免疫功能受损的人。多重耐药铜绿假单胞菌 已被疾病控制和预防中心确定为严重威胁。铜绿假单胞菌致病原因 在各种临床环境和宿主中与医疗保健相关的感染，但在 囊性纤维化(CF)患者。我们对使用称为PPMO的反义分子作为 这些感染的潜在治疗方法。这些分子阻断信使rna，防止形成 目标蛋白。我们已经证明，PPMO可以被用来靶向对以下过程至关重要的基因 假单胞菌的生长，如acpP、lpxC或rpsJ。我们的研究表明，阻断这些蛋白质对于 假单胞菌在体外生长。我们还表明，这些PPMO提高了感染的小鼠的存活率 假单胞菌。对于这个项目，我们建议在一个更大的集合中进一步描述我们的主要PPMO 假单胞菌分离株，对抗生素敏感和多重耐药。此外，还将进行功效研究 在肺炎和血流感染的两种模型中都进行了检测。这一过程将产生2-4个PPMO 将接受进一步的临床前测试，包括毒性、耐药性、药效学和药动学 学习。在拟议项目结束时，一名首席PPMO将完成必要的临床前测试 用于向FDA提交IND。这是一种开发新型抗生素的创新方法，特别是对P. 铜绿假单胞菌可能有助于扩大用于治疗的有效抗生素的日益减少的类别 这些严重的、危及生命的感染。

PROJECT SUMMARY/ABSTRACT Gram-negative pathogens are becoming increasingly resistant to many antimicrobials. Furthermore, the pipeline for new antibiotics is small and new therapies are urgently needed. This can be especially problematic in patients who suffer from chronic infections or are immunocompromised. Multidrug-resistant Pseudomonas aeruginosa has been identified by the Centers for Disease Control and Prevention as a serious threat. P. aeruginosa causes healthcare associated infections in a variety of clinical settings and hosts, but is particularly devastating in patients with cystic fibrosis (CF). We have been interested in using antisense molecules called PPMOs as potential therapeutics in these infections. These molecules block messenger RNA and prevent the formation of the target protein. We have demonstrated that PPMOs can be used to target genes that are essential for Pseudomonas to grow, such as acpP, lpxC or rpsJ. We showed that blocking these proteins are essential for Pseudomonas to grow in vitro. We also showed that these PPMOs improve survival in mice that were infected with Pseudomonas. For this project, we propose to further characterize our lead PPMOs in a larger collection of Pseudomonas isolates, both antibiotic-sensitive and multidrug-resistant. In addition, efficacy studies will be performed in both models of pneumonia and bloodstream infection. This process will result in 2-4 PPMOs that will undergo further pre-clinical testing including toxicity, resistance, pharmacodynamic and pharmacokinetic studies. By the end of the proposed project, a lead PPMO will have undergone the needed pre-clinical testing for IND submission to the FDA. This innovative approach to developing novel antibiotics, particularly for P. aeruginosa, could help expand the increasingly shrinking classes of effective antibiotics that are used to treat these severe, life-threatening infections.