Function of MVFR in Pseudomonas Aeruginosa Virulence

MVFR 在铜绿假单胞菌毒力中的作用

基本信息

批准号：
8528902
负责人：
LAURENCE G RAHME
金额：
$ 47.5万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-21 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8528902
关键词：
Acute Animal Model Animals Anthranilic Acids Anti-Infective Agents Antibiotic Resistance Antibiotics Attenuated Bacteria Bacterial Infections Benzamides Benzimidazoles Binding Biochemical Burn injury Bypass Cells Chemicals Chronic Communication DNA DNA Binding Data Development Emerging Communicable Diseases Feedback Figs - dietary Funding Generations Genes Glossary Goals Gram-Negative Bacteria Growth Health Hospitals Human Incidence Infection Intensive Care Units Laboratories Lead Ligand Binding Ligands Liquid Chromatography Magnetic Resonance Imaging Mammals Mediating Metabolism Microbe Modeling Molecular Genetics Molecular Target Molecular Weight Multi-Drug Resistance Mus Operon Oxides Pathogenesis Pathway interactions Play Population Population Density Process Production Property Protein Region Pseudomonas aeruginosa Public Health Refractory Regulation Regulon Relapse Research Resistance Role Signal Pathway Signal Transduction Spectrometry Structure Structure-Activity Relationship System Testing Text Therapeutic Virulence Virulence Factors Work absorption antimicrobial antimicrobial drug benzimidazole biodefense combat commensal microbes density drug discovery genetic analysis high throughput screening improved in vivo inhibitor/antagonist innovation insight intercellular communication iron oxide killings liquid chromatography mass spectrometry mutant novel novel strategies pathogen pressure prevent promoter public health relevance quorum sensing small molecule transcription factor

项目摘要

DESCRIPTION (provided by applicant): New approaches to antimicrobial drug discovery are urgently needed to combat untreatable infections caused by antibiotic resistant or persistent, antibiotic tolerant, bacteria. The long-term goal of this proposal is to develop novel therapy options that may prevent or reduce the complications of human bacterial acute and chronic or relapsing infections, and that could serve as alternatives or adjuncts to antibiotics. To achieve a paradigm shift in antimicrobial therapy, we propose to develop inhibitors of bacterial signaling pathways that control bacterial virulence and antibiotic tolerance mechanisms. Population density-dependent signaling, generally referred to as quorum sensing (QS), is one such mechanism. QS regulates multiple aspects of virulence. It is important for the development of acute infections and as recently discovered for the formation of antibiotic-tolerant cell populations, a process underlying pathogen persistence in chronic infections. This renewal application will test the hypothesis that novel inhibitors of QS, previously identified by us, can lead to the development of highly potent compounds with anti-infective activity in vivo. We will test this hypothesis by employing Pseudomonas aeruginosa, a recalcitrant Gram-negative bacterium that defies eradication by antibiotics and exemplifies current problematic pathogens in hospitals and intensive care units. In the last funding cycle, we demonstrated that P. aeruginosa pathogenesis can be disrupted in vivo by pharmacologically interfering with the multiple virulence factor regulator (MvfR) regulon, a component of QS circuitry that controls virulence and, as we recently discovered, the formation of antibiotic tolerant cells. We elucidated the mechanism of MvfR regulon activation and identified several small chemical compounds that inhibit the MvfR transcription factor and/or interfere with MvfR regulon activity in vivo. Using these chemical compounds, we will test our hypothesis through three Specific Aims: 1) To study the mechanisms of action of the candidate compounds using biochemical, mass spectrometric, and molecular genetic analyses. 2) To improve the most potent QS inhibitors through structure-activity relationship (SAR) studies. 3) To validate the in vivo efficacy of SAR-improved inhibitors in attenuating P. aeruginosa infection in suitable animal models. QS signaling circuits are evolutionarily conserved and play central roles in modulating virulence mechanisms in many different human pathogens. Therefore, by selectively interfering with QS, our data should yield paradigmatic insights that will be generally relevant for the development of new classes of anti-infectives that could limit development of multi-drug resistance and antibiotic tolerance in bacterial pathogens, while preserving beneficial commensal bacteria. Moreover, the study and inhibition of mechanisms involved in antibiotic tolerance could have a major impact on elucidating this unresolved phenomenon, as well as enabling the discovery of the first probe compounds targeted against antibiotic tolerant cells.

描述（由申请人提供）：迫切需要进行抗菌药物发现的新方法，以应对由抗生素耐药或持续性，抗生素耐受性的细菌引起的不可治疗的感染。该提案的长期目标是开发新的治疗选择，以防止或减少人类细菌急性，慢性或复发感染的并发症，并可以用作抗生素的替代或辅助性。为了实现抗菌治疗的范式转移，我们建议开发控制细菌毒力和抗生素耐受性机制的细菌信号传导途径的抑制剂。总体密度依赖性信号传导通常称为Quorum Sensing（QS），就是一种机制。 QS调节毒力的多个方面。这对于急性感染的发展很重要，并且由于最近发现的抗生素耐耐药细胞群体而言，这是慢性感染中病原体持久性的过程。这种更新应用将检验以下假设：我们以前鉴定出的QS的新型抑制剂可以导致在体内具有抗感染活性的高效化合物的发展。我们将通过采用铜绿假单胞菌来检验这一假设，这是一种顽固的革兰氏阴性细菌，抗生素消除了根除，并体现了医院和重症监护病房中当前的有问题病原体。在最后一个融资周期中，我们证明了铜绿假单胞菌的发病机理可以通过药理学干扰多重毒力因子调节剂（MVFR）调节剂来破坏体内，这是控制毒力的QS电路的一个成分，并且正如我们最近发现的那样，正如我们最近发现的那样，抗生素耐受性细胞的形成。我们阐明了MVFR调节活化的机理，并鉴定了几种抑制MVFR转录因子和/或干扰MVFR调节活性的小型化合物。使用这些化合物，我们将通过三个特定目的测试我们的假设：1）使用生化，质谱和分子遗传分析研究候选化合物的作用机理。 2）通过结构活性关系（SAR）研究改善最有效的QS抑制剂。 3）验证在合适的动物模型中，验证SAR-改良抑制剂在减弱铜绿假单胞菌感染中的体内功效。 QS信号通电在进化上是保守的，并且在调节许多不同人类病原体中的毒力机制中起着核心作用。因此，通过选择性干扰QS，我们的数据应产生范式的见解，这些见解通常与新的抗诊所的开发有关，这些抗探视剂可能会限制细菌病原体中多药耐药性和抗生素耐受性的发展，同时保留有益的儿童细菌。此外，对抗生素耐受性涉及的机制的研究和抑制可能对阐明这种未解决的现象产生重大影响，并能够发现针对抗生素耐受细胞的第一个探针化合物。

项目成果

期刊论文数量（15）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（1）

A quorum sensing small volatile molecule promotes antibiotic tolerance in bacteria.

群体感应小挥发性分子可促进细菌的抗生素耐受性。

DOI：
10.1371/journal.pone.0080140
发表时间：
2013
期刊：
PloS one
影响因子：
3.7
作者：
Que,Yok-Ai;Hazan,Ronen;Strobel,Benjamin;Maura,Damien;He,Jianxin;Kesarwani,Meenu;Panopoulos,Panagiotis;Tsurumi,Amy;Giddey,Marlyse;Wilhelmy,Julie;Mindrinos,MichaelN;Rahme,LaurenceG
通讯作者：
Rahme,LaurenceG

Considerations and caveats in anti-virulence drug development.

DOI：
10.1016/j.mib.2016.06.001
发表时间：
2016-10
期刊：
Current opinion in microbiology
影响因子：
5.4
作者：
Maura D;Ballok AE;Rahme LG
通讯作者：
Rahme LG

A method for high throughput determination of viable bacteria cell counts in 96-well plates.

DOI：
10.1186/1471-2180-12-259
发表时间：
2012-11-13
期刊：
BMC microbiology
影响因子：
4.2
作者：
Hazan R;Que YA;Maura D;Rahme LG
通讯作者：
Rahme LG

Molecular Insights into Function and Competitive Inhibition of Pseudomonas aeruginosa Multiple Virulence Factor Regulator.

DOI：
10.1128/mbio.02158-17
发表时间：
2018-01-16
期刊：
mBio
影响因子：
6.4
作者：
Kitao T;Lepine F;Babloudi S;Walte F;Steinbacher S;Maskos K;Blaesse M;Negri M;Pucci M;Zahler B;Felici A;Rahme LG
通讯作者：
Rahme LG

Assessing Pseudomonas aeruginosa Persister/antibiotic tolerant cells.