Gene silencing therapeutics for chronic infections in cystic fibrosis

囊性纤维化慢性感染的基因沉默疗法

• 批准号: 9248236
• 负责人: David Elihu Greenberg
• 金额: $ 36.08万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-03-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9248236
• 关键词: Acyl Carrier Protein; Anabolism; Animals; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Antisense Technology; Biological Assay; Burkholderia; Burkholderia cepacia complex; Carrier Proteins; Cell Wall; Chemistry; Chronic; Clinical; Cystic Fibrosis; Databases; Deterioration; Development; Disease; Drug resistance; Escherichia coli; Essential Genes; Future; Gene Silencing; Gene Targeting; Genes; Goals; Growth; Human; In Vitro; Infection; Lactamase; Lead; Lipopolysaccharides; Lung; Medical; Messenger RNA; Methods; Microbial Biofilms; Minimum Inhibitory Concentration measurement; Modeling; Morbidity - disease rate; Multi-Drug Resistance; Mus; Pathway interactions; Patients; Peptides; Peptidoglycan; Pharmaceutical Preparations; Phase; Physiological; Play; Predisposition; Proteins; Pseudomonas; Pseudomonas aeruginosa; Public Health; Resistance; Respiratory physiology; Role; Salmonella; Structure; Syndrome; Testing; Therapeutic; Therapeutic Agents; Work; antimicrobial; bacterial resistance; clinical development; cystic fibrosis patients; design; efficacy study; efflux pump; fatty acid biosynthesis; fighting; improved; in vivo; interest; mortality; mouse model; multi-drug resistant pathogen; novel; novel therapeutics; pathogen; phosphorodiamidate morpholino oligomer; resistance gene; resistance mechanism; response; therapeutic development; therapeutic target

PROJECT SUMMARY/ABSTRACT Gram-negative pathogens are becoming increasingly resistant to currently used antimicrobials. Furthermore, the pipeline for new antibiotics is slim and new therapies are urgently needed. This can be especially problematic in patients who suffer from chronic infections. Examples of inherently drug resistant Gram-negative pathogens are Pseudomonas aeruginosa and the Burkholderia cepacia complex (Bcc). These pathogens cause disease in a variety of settings, but are particularly devastating in patients with cystic fibrosis (CF). Chronic P. aeruginosa or Bcc infection can lead to both progressive pulmonary decline, as well as in Bcc infection, rapid deterioration in lung function referred to as “cepacia syndrome.” We have been interested in using antisense molecules called PPMOs as potential therapeutics in these infections. These molecules block messenger RNA and block the formation of protein. We have demonstrated that PPMOs can be used to target genes that are essential for these pathogens to grow, specifically the gene acpP (that encodes the protein acyl carrier protein). We showed that blocking this protein is essential for Burkholderia and Pseudomonas to grow in vitro. We also showed that this PPMO improves survival in mice that were infected with Burkholderia. For this project, we propose to find other essential gene targets in these pathogens that can be blocked with PPMOs, and perform studies to evaluate the most promising compounds ability to improve survival in mouse models of infection. In the first phase of the project, we will target pathways that have been shown in other Gram-negative pathogens to be essential for growth. These will include genes important in the development of the cell wall and associated structures (lipopolysaccharide and peptidoglycan biosynthesis) as well as fatty acid biosynthesis (i.e. acyl carrier protein). In addition, we will try to block the genes that play a role in antibiotic resistance to see if we can restore activity of currently existing antibiotics. The second phase of the project will be to evaluate lead PPMO candidates in mouse models of infection. We will use the PPMOs as treatments in pulmonary models of infection, assessing both systemic and pulmonary delivery of the compound. By the conclusion of this study, we hope to have promising novel treatments for two of the most devastating pathogens in CF that could advance to further clinical development. This work has implications not only for these infections in CF, but for other medically important Gram-negative pathogens.

项目摘要/摘要 革兰氏阴性病原体正越来越抗当前使用的抗菌药物。此外， 新抗生素的管道很苗条，迫切需要新的疗法。特别是 患有慢性感染的患者有问题。固有耐药革兰氏阴性的示例 病原体是铜绿假单胞菌和Burkholderia cepacia Complex（BCC）。这些病原体 在各种环境中引起疾病​​，但在囊性纤维化患者（CF）的患者中尤其是毁灭性的。 慢性铜绿假单胞菌或BCC感染可导致肺的进行性下降，也可以导致BCC 感染，肺功能的快速定义称为“肝综合征”。我们一直对 在这些感染中，使用称为PPMO的反义分子作为潜在的治疗。这些分子阻塞 Messenger RNA并阻止蛋白质的形成。我们已经证明PPMO可用于靶向 这些病原体生长至关重要的基因，特别是基因ACPP（它编码蛋白 载体蛋白）。我们表明，阻断该蛋白对于Burkholderia和Pseudomonas生长至关重要 体外。我们还表明，这种PPMO改善了被Burkholderia感染的小鼠的生存。为了 这个项目，我们建议在这些病原体中找到其他可能被阻止的基因靶标 PPMO，并进行研究以评估提高小鼠生存的最有希望的化合物的能力 感染模型。在项目的第一阶段，我们将针对其他已显示的途径 革兰氏阴性病原体对于生长至关重要。这些将包括在开发中很重要的基因 细胞壁和相关结构（脂多糖和辣椒生物合成）以及脂肪 酸性生物合成（即酰基载体蛋白）。此外，我们将尝试阻止在抗生素中发挥作用的基因 抵抗力查看我们是否可以恢复当前现有抗生素的活性。该项目的第二阶段将 要评估在感染小鼠模型中铅PPMO候选物。我们将使用PPMO作为治疗 感染的肺模型，评估化合物的全身和肺部递送。由 这项研究的结论，我们希望对最毁灭性的两个有望有新的治疗 CF中的病原体可以发展为进一步的临床发育。这项工作不仅对 这些感染在CF中，但对于其他重要的革兰氏阴性病原体。

项目成果

Peptide-Conjugated Phosphorodiamidate Morpholino Oligomers Retain Activity against Multidrug-Resistant Pseudomonas aeruginosa In Vitro and In Vivo.

• DOI: 10.1128/mbio.02411-20
• 发表时间: 2021-01-12
• 期刊: mBio
• 影响因子: 6.4
• 作者: Moustafa DA;Wu AW;Zamora D;Daly SM;Sturge CR;Pybus C;Geller BL;Goldberg JB;Greenberg DE
• 通讯作者: Greenberg DE