Antimicrobial peptides from the microbiome as modulators of bacterial virulence

来自微生物组的抗菌肽作为细菌毒力的调节剂

• 批准号: RGPIN-2017-05322
• 负责人: Duperthuy, Marylise
• 金额: $ 1.89万
• 依托单位: Université de Montréal
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=649791
• 关键词: Antimicrobial; peptides; microbiome; modulators; bacterial

The gut microbiome has been intensively studied lately because of its fundamental role in human health. However, the molecular dialogue between the bacteria from the microbiome is still poorly understood. My research focus on understanding this dialogue in a cooperative or a competitive bacterial relationship. ******The bacteria from the microbiome are producing antimicrobial peptides (AMPs), which have an important role in regulating the microbial community. The aim of this proposal is to determine if AMPs produced by the microbiome can modulate the expression and secretion of proteins involved in virulence and resistance in V. cholerae. Therefore, our specific objectives are to: ***i) Identify and select bacteria from the microbiome that produce AMP(s) active against V. cholerae. Two approaches will be considered. A random approach consisting of the screening of a bacterial collection from the microbiome for their anti-Vibrio activity, followed by an identification of the producer and a purification of the AMPs. The second approach consist on the selection of the AMPs produced by bacteria from the microbiome based on the available literature, eventually followed by a cloning and purification.***ii) Determine how the AMPs from the microbiome affect the expression and secretion of proteins from V. cholerae. A global proteomic approach will be performed to identify the protein differentially expressed in presence of AMPs. A construction of the mutants of the genes coding for the identified effectors will follow for the identification of the role of the effector proteins in antimicrobial peptide resistance and in virulence using animal models.***iii) Identify the risk of developing a resistance to the AMPs from the microbiome, by cultivating different intestinal pathogens with low concentration of the AMPs from the microbiome.******This work will greatly improve our knowledge of bacteria-bacteria communication in the gut at the molecular level. The novelty resides in considering AMPs as molecules that can subvert the signalling pathways of the neighbouring bacteria, including pathogens, therefore modulating their ability to adapt to their environment.

肠道微生物组由于其在人类健康中的重要作用，最近得到了深​​入研究。然而，微生物组中细菌之间的分子对话仍然知之甚少。我的研究重点是理解合作或竞争细菌关系中的这种对话。 ******微生物群中的细菌正在产生抗菌肽 (AMP)，它在调节微生物群落方面发挥着重要作用。该提案的目的是确定微生物组产生的 AMP 是否可以调节霍乱弧菌毒力和耐药性相关蛋白质的表达和分泌。因此，我们的具体目标是：***i) 从微生物组中识别和选择能够产生对霍乱弧菌具有活性的 AMP 的细菌。将考虑两种方法。一种随机方法，包括从微生物组中筛选细菌集合的抗弧菌活性，然后鉴定生产者并纯化 AMP。第二种方法包括根据现有文献从微生物组中选择细菌产生的 AMP，最终进行克隆和纯化。***ii) 确定微生物组中的 AMP 如何影响微生物组中蛋白质的表达和分泌霍乱弧菌。将采用全局蛋白质组学方法来鉴定在 AMP 存在下差异表达的蛋白质。随后将构建编码已识别效应子的基因突变体，以便使用动物模型来鉴定效应蛋白在抗菌肽耐药性和毒力中的作用。***iii) 识别对抗菌肽产生耐药性的风险来自微生物组的 AMP，通过使用低浓度的来自微生物组的 AMP 培养不同的肠道病原体。******这项工作将极大地提高我们对肠道中细菌与细菌在分子水平上通讯的认识。新颖之处在于将 AMP 视为可以破坏邻近细菌（包括病原体）信号传导途径的分子，从而调节它们适应环境的能力。