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.

项目概要/摘要 革兰氏阴性病原体对目前使用的抗菌药物的耐药性越来越强。 新抗生素的研发渠道很窄，因此迫切需要新的疗法。 患有慢性感染的患者会遇到问题 固有耐药的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菌复合体（Bcc）。 在多种情况下都会引起疾病，但对囊性纤维化 (CF) 患者的破坏力尤其严重。 慢性铜绿假单胞菌或 Bcc 感染可导致进行性肺功能衰退以及 Bcc 我们一直对感染、肺功能迅速恶化（称为“cepacia 综合征”）感兴趣。 使用称为 PPMO 的反义分子作为这些感染的潜在疗法。 我们已经证明 PPMO 可用于靶向。 这些病原体生长所必需的基因，特别是 acpP 基因（编码酰基蛋白） 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。 我们还表明，这种 PPMO 可以提高感染伯克霍尔德氏菌的小鼠的存活率。 在这个项目中，我们建议在这些病原体中寻找可以用以下方法阻断的其他重要基因靶标： PPMO，并进行研究以评估最有希望的化合物提高小鼠存活率的能力 在该项目的第一阶段，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，其中包括对发育至关重要的基因。 细胞壁和相关结构（脂多糖和肽聚糖生物合成）以及脂肪 此外，我们将尝试阻断在抗生素中发挥作用的基因。 该项目的第二阶段将研究耐药性，看看我们是否能够恢复现有抗生素的活性。 我们将在小鼠感染模型中评估主要 PPMO 候选药物。 肺部感染模型，评估化合物的全身和肺部递送。 根据这项研究的结论，我们希望针对两种最具破坏性的疾病找到有希望的新疗法 这项工作不仅对 CF 中的病原体具有重要意义，而且可能会推动进一步的临床开发。 这些感染主要针对CF，但也针对其他医学上重要的革兰氏阴性病原体。

项目成果

AcrAB-TolC Inhibition by Peptide-Conjugated Phosphorodiamidate Morpholino Oligomers Restores Antibiotic Activity in Vitro and in Vivo.

肽缀合的磷酸二酰胺吗啉代低聚物对 AcrAB-TolC 的抑制可恢复体外和体内的抗生素活性。

• DOI: 10.1021/acsinfecdis.9b00123
• 发表时间: 2019-05-23
• 期刊: ACS infectious diseases
• 影响因子: 5.3
• 作者: Carolyn R. Sturge;Christina F Felder;R. Pifer;Christine Pybus;R. Jain;B. Geller;D. Greenberg
• 通讯作者: D. Greenberg

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

肽缀合的磷酰二胺吗啉代低聚物在体外和体内均保留针对多重耐药铜绿假单胞菌的活性。

• 发表时间: 2021
• 影响因子: 6.4
• 作者: Moustafa, Dina A;Wu, Ashley W;Zamora, Danniel;Daly, Seth M;Sturge, Carolyn R;Pybus, Christine;Geller, Bruce L;Goldberg, Joanna B;Greenberg, David E
• 通讯作者: Greenberg, David E