Pursuing Antivirulence of MRSA in Penicillium sp. with droplet surface-sampling-liquid microjunction-probe

寻求青霉属 MRSA 的抗毒力。

• 批准号: 9314217
• 负责人: Diana Kao
• 金额: $ 3.05万
• 依托单位: UNIVERSITY OF NORTH CAROLINA GREENSBORO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9314217
• 关键词: Adjuvant; Aggressive behavior; Anabolism; Antibiotics; Attention; Attenuated; Bacteria; Binding; Data; Defect; Development; Diagnosis; Drug Targeting; Drug resistance; Environment; Epigenetic Process; Evaluation; Excision; Faculty; Goals; Growth; Hospitals; In Situ; Infection; Infectious Skin Diseases; Investigation; Lead; Liquid substance; Mass Spectrum Analysis; Methicillin Resistance; Methods; Milk Thistle; Monitor; Morphology; Pathway interactions; Patients; Pattern; Penicillium; Periodicity; Pharmaceutical Preparations; Production; Regulator Genes; Research; Resolution; Sampling; Series; Signal Transduction; Skin Tissue; Soft Tissue Infections; Staphylococcus aureus; Stress; Surface; System; Techniques; Testing; Variant; Virulence; analog; attenuation; cell growth; data acquisition; drug candidate; drug testing; endophytic fungi; fungus; improved; in vivo; methicillin resistant Staphylococcus aureus; microorganism; mouse model; pathogen; programs; quorum sensing; response; success

PROJECT SUMMARY More potent drugs that seek to eradicate the more drug resistant forms of Methicillin-resistant Staphyloccocus aureus (MRSA), a devastating skin infection, only promotes further drug resistance. An alternate treatment is necessary, and antivirulence could be that treatment. By inhibiting antivirulence factors in the accessory gene regulator (agrA), the quorum sensing pathway is quenched. This suppresses aggression of the pathogen, which allows the host to clear the infection. ω-Hydroxyemodin has shown success inhibiting MRSA skin and soft tissue infections in an in vivo mouse model through this pathway by binding to agrA, the response regulator. Because further testing is necessary before a drug can reach hospitals, a greater supply as well as alternate analogues is necessary. This project seeks to not only increase the supply of ω-hydroxyemodin from Penicillium restrictum, a known producer of the compound, but also examine the other faculties of the microorganism that may produce analogues that show equal or greater activity than ω-hydroxyemodin. By changing the culture conditions and retaining the same fungus, the biosynthetic pathway for polyhydroxyanthraquinones will remain but the types of polyhydroxyanthraquinones produced will change. Profiles of each condition will be rapidly accessed with droplet-liquid microjunction-surface sampling probe (droplet-LMJ-SSP). The profiles can be compared not only to determine optimal growing conditions for the production of the key active compound but also significant differences in metabolite profile to merit further investigation. These compounds will be tested for inhibition of MRSA and characterized via high resolution mass spectrometry and a suite of NMR techniques. This study combines the one strain-many compounds approach to stress and change the secondary metabolite production of the fungus, droplet-liquid microjunction- surface sampling probe to examine the secondary metabolites in situ, and chemometrics and mass defect filtering to characterize and analyze secondary metabolite profiles.

项目摘要 寻求根除耐甲氧西林葡萄球菌的耐药性更强的药物 金黄色葡萄球菌（MRSA），一种破坏性的皮肤感染，只会促进进一步的耐药性。另一种治疗方法是 必要的，和抗病力可能是治疗。通过抑制辅助基因中的抗毒力因子 调节子（agrA），群体感应途径被淬灭。这抑制了病原体的侵略， 让宿主清除感染ω-羟基大黄素已显示出成功抑制MRSA皮肤， 通过与agrA结合，通过该途径在体内小鼠模型中进行软组织感染， 调节器因为在药物到达医院之前需要进行进一步的测试，所以更多的供应以及 替代的类似物是必要的。该项目不仅旨在增加ω-羟基大黄素的供应， 限制青霉菌，一种已知的化合物生产者，也检查了细胞的其他功能。 可以产生类似物的微生物，所述类似物显示出与ω-羟基大黄素相等或更大的活性。通过 改变培养条件并保留相同的真菌， 多羟基蒽醌将保留，但产生的多羟基蒽醌的类型将改变。 每个条件下的轮廓将快速访问与液滴-液体微界面-表面采样探针 （液滴-LMJ-SSP）。可以比较所述分布，不仅以确定所述植物的最佳生长条件， 关键活性化合物的生产，但代谢物谱的显著差异，值得进一步研究。 调查将测试这些化合物对MRSA的抑制作用，并通过高分辨率进行表征 质谱法和一套核磁共振技术。这项研究结合了一个菌株-许多化合物 胁迫和改变真菌次级代谢产物产生的方法，液滴-液体微连接- 表面取样探针原位检测次级代谢产物，化学计量学和质量亏损 过滤以表征和分析次级代谢产物谱。