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.

项目总结/摘要 革兰氏阴性病原体对目前使用的抗菌剂的耐药性越来越强。此外，委员会认为， 新抗生素的生产线很窄，迫切需要新的治疗方法。这可能是特别 对慢性感染的患者来说是个问题。固有耐药革兰氏阴性菌的例子 病原体是铜绿假单胞菌和洋葱伯克霍尔德氏菌复合体（Bcc）。这些病原体 在各种情况下引起疾病，但在囊性纤维化（CF）患者中尤其具有破坏性。 慢性铜绿假单胞菌或Bcc感染可导致进行性肺功能下降，以及Bcc 感染后，肺功能迅速恶化称为“洋葱综合征”。我们一直对 使用称为PPMO的反义分子作为这些感染的潜在治疗剂。这些分子阻断了 信使RNA并阻止蛋白质的形成。我们已经证明，PPMOs可以用于靶向 这些基因是这些病原体生长所必需的，特别是基因acpP（编码蛋白质酰基 载体蛋白）。我们发现阻断这种蛋白质对于伯克霍尔德氏菌和假单胞菌的生长至关重要 体外我们还表明，这种PPMO提高了感染伯克霍尔德氏菌的小鼠的存活率。为 在这个项目中，我们建议在这些病原体中找到其他可以被阻断的必需基因靶点。 PPMO，并进行研究，以评估最有前途的化合物改善小鼠存活率的能力 感染的模型。在该项目的第一阶段，我们将针对已在其他研究中显示的途径， 革兰氏阴性病原体对生长至关重要。这些基因将包括在以下发育过程中重要的基因： 细胞壁和相关结构（脂多糖和肽聚糖生物合成）以及脂肪 酸生物合成（即酰基载体蛋白）。此外，我们将尝试阻断在抗生素中发挥作用的基因， 看看我们是否能恢复现有抗生素的活性。该项目的第二阶段将 在小鼠感染模型中评估PPMO候选药物。我们将使用PMOs作为治疗方法， 肺部感染模型，评估化合物的全身和肺部递送。由 在这项研究的结论中，我们希望有有希望的新疗法来治疗两种最具破坏性的疾病。 CF中的病原体，可以推进到进一步的临床开发。这项工作不仅对 这些感染CF，但对于其他医学上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