Identification of specific metabolites in mycolactone producing mycobacteria and Buruli ulcer infection: diagnostic biomarkers through metabolomic

产生分枝内酯的分枝杆菌和布鲁里溃疡感染中特定代谢物的鉴定：通过代谢组学诊断生物标志物

• 批准号: MR/P024416/1
• 负责人: Julian Griffin
• 金额: $ 28.45万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FP024416%2F1
• 关键词: Identification; specific; metabolites; mycolactone; producing

Buruli ulcer is a severe, slow progressing skin infection which destroys tissue and is caused by bacteria found in the environment called Mycobacterium ulcerans. The disease is classified as a neglected tropical disease despite being described in over 30 countries including Ghana, Togo, Benin, Côte d'Ivoire, Cameroon, Mexico, Papua New Guinea, South East Asia and Australia, and mostly affects individuals in rural communities living in infected areas. The World Health Organisation's Global Buruli Ulcer Initiative has identified the development of simple diagnostic tools as one of the priority areas in the control of the disease. Metabolomics is a growing field in analytical science and biology which seeks to identify novel biomarkers that can distinguish between diseased and non-diseased individuals. In a pilot study, funded under the Cambridge-Africa Partnership for Research Excellence (CAPREx), we sought to determine the best types of sample for the identification of metabolites from the bacteria that can be found only in Buruli ulcer patients. We collected tissue biopsy, swabs and fine needle aspirates from 28 Buruli ulcer confirmed patents and 21 patients with tropical ulcers that were not Buruli ulcer. The samples were pre-processed at the Department of Biochemistry, Cell and Molecular Biology at the University of Ghana, to extract the required components for gas chromatography-mass spectrometry (GC-MS) at the University of Cambridge. We identified tissue biopsy and fine needle aspirates as the best specimen for the identification of metabolites in preference to lesion swabs. Interestingly, metabolites identified in both groups of patients included cadaverine, putrescine, pinitol, palmitate, naphthalene, chlopyrifos and oxaspiro. The former two metabolites are interesting because they classify all the samples as containing degenerating tissue. The fatty acid palmitate is a common metabolite present in human tissue. Pinitol and the later three identifies metabolites are chemical residues that may have been present in the treatment poultice used as unorthodox remedies by the patients. In this proposal, we are seeking to further this pilot study, building on our observations and collecting the tissue biopsies and fine needle aspirates from a larger patient sample group to allow us to perform a more detailed analysis of the compounds found within the bacteria and also within infected tissue. We also intend to expand the scope of the biological samples from affected individuals by including urine samples in our analysis which may provide a more convenient biofluid to sample. To gain more insight into the novelty of the biomarkers we identify, we will include another phase of experimentation which involves the characterization of the compounds found in M. ulcerans and other similar bacteria strains that infect people in Ghana including M. pseudishottsii, M. liflandii and M. marinum DL. Data generated from this project will be important in the identification of new diagnostics and also give us insight to the biology of the mycobacterium during infection which may lead to better prospects for future treatments.

布鲁里溃疡是一种严重的，缓慢进展的皮肤感染，破坏组织，是由环境中发现的称为溃疡分枝杆菌的细菌引起的。尽管在加纳、多哥、贝宁、科特迪瓦、喀麦隆、墨西哥、巴布亚新几内亚、东南亚和澳大利亚等30多个国家都有描述，但这种疾病被列为一种被忽视的热带疾病，主要影响生活在受感染地区的农村社区的个人。世界卫生组织的全球布鲁里溃疡倡议已将开发简单的诊断工具确定为控制该疾病的优先领域之一。代谢组学是分析科学和生物学中一个不断发展的领域，旨在鉴定可以区分患病和非患病个体的新型生物标志物。在剑桥-非洲卓越研究伙伴关系（CAPREx）资助的一项试点研究中，我们试图确定用于鉴定仅在布鲁里溃疡患者中发现的细菌代谢物的最佳样品类型。我们收集了28例布鲁里溃疡确诊患者和21例非布鲁里溃疡的热带溃疡患者的组织活检、拭子和细针抽吸物。加纳大学生物化学、细胞和分子生物学系对样本进行了预处理，以提取剑桥大学气相色谱-质谱法（GC-MS）所需的成分。我们确定组织活检和细针抽吸物是鉴定代谢物的最佳标本，而不是病变拭子。有趣的是，在两组患者中鉴定的代谢物包括尸胺、腐胺、松醇、棕榈酸酯、萘、毒死蜱和氧杂螺。前两种代谢物是有趣的，因为它们将所有样品分类为含有退化组织。脂肪酸棕榈酸酯是人体组织中常见的代谢物。Pinitol和后三种确定的代谢物是可能存在于患者用作非正统疗法的治疗膏药中的化学残留物。在这项提案中，我们正在寻求进一步开展这项试点研究，建立在我们的观察基础上，并从更大的患者样本组中收集组织活检和细针抽吸物，以使我们能够对细菌内和感染组织内发现的化合物进行更详细的分析。我们还打算扩大受影响个体的生物样本的范围，将尿液样本纳入我们的分析中，这可能会提供更方便的生物流体样本。为了更深入地了解我们鉴定的生物标志物的新奇，我们将包括另一个实验阶段，该阶段涉及M中发现的化合物的表征。溃疡和其他类似的细菌菌株感染加纳的人，包括M。pseudishottsii、假丝毛孢M. liflandii和M.海百合从该项目产生的数据将在识别新的诊断方法中发挥重要作用，并使我们了解感染期间分枝杆菌的生物学，这可能会为未来的治疗带来更好的前景。

项目成果

How to bridge metabolomics and genomics.

如何连接代谢组学和基因组学。

• DOI: 10.1016/j.biocel.2017.11.001
• 发表时间: 2017
• 期刊: The international journal of biochemistry & cell biology
• 作者: Atzori L