A genome-scale census of virulence factors in the major mould pathogen of human lungs

人类肺部主要霉菌病原体毒力因子的基因组规模普查

• 批准号: MR/M02010X/1
• 负责人: Elaine Bignell
• 金额: $ 94.29万
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FM02010X%2F1
• 关键词: genome; scale; census; virulence; factors

Infectious diseases caused by fungi are a worldwide problem causing at least as many deaths as malaria and tuberculosis. For infections caused by spore-forming moulds there is only one available class of oral drug, the azoles. Azole resistance amongst clinical mould isolates is increasing and one possible cause of this problem is the widespread use of azoles in the environment as agricultural fungicides. There is therefore a desperate need to develop new antifungal treatments.Fungal particles (or spores), which are continually present in the air that we breathe, are able to survive in the human lung. Usually, this occurs when the immune system of the infected individual is not functioning optimally. Always, it involves the germination of the fungal spore (rather like a plant seed) and growth of long fungal cells called hyphae. Hyphae penetrate and dissolve lung tissues by secreting enzymes. The major fungal pathogen in the air we breathe is called Aspergillus fumigatus and people affected by cancer, or requiring organ transplants, are particularly at risk of fatal infections caused by it. Unfortunately our lack of understanding of A. fumigatus, and the technical difficulties encountered when working with it, has limited progress in this field. We do not fully understand why Aspergillus is able to survive inside the lung environment. We urgently need more basic information about this pathogen so we can begin to design future therapies. When scientists try to figure out how cellular processes work, a first port of call is often transcription factors. Transcription factors control the activity of many genes simultaneously so they command great power over cellular processes. If a transcription factor is important for a particular process, one will then look at which genes it regulates and this will give clues to the biology underlying that process. If a transcription factor is not involved then it, and all of the genes it regulates, can be eliminated from the investigation. By asking which transcription factors control the ability of A. fumigatus to cause disease, we can achieve a global view of the regulatory network which drives infection. We intend to couple this approach to a state of the art DNA sequencing technology to gain maximal insight on A. fumigatus pathogenicity, with minimal usage of mice. We have searched the A. fumigatus genome for transcription factor genes and created a collection of 401 A. fumigatus mutants, each one lacking a transcription factor. To ask which of the transcription factors is important for mammalian lung infection we will infect mice with the collection of mutants and see which are unable to survive inside the lung. We have devised a method which will allow us to test many mutants at the same time. This involves infecting all of the mutants into a single mouse, and after sacrifice, sequencing the DNA extracted from the lung to tell us which mutants have managed to survive. We will use this method to reveal all of the transcription factors driving pathogenicity (P-TFs) in A. fumigatus. Thus the lowest possible number of animals will be utilised. When we have identified all of the P-TFs we will then work to identify the genes which are regulated by each P-TF. In order to do this we will use two approaches. First we will chemically fix the transcription factor to A. fumigatus DNA to identify the binding sites of each transcription factor. Second we will look at gene expression in mutants lacking P-TFs and work out which genes become deregulated. The success of this investigation will 1) Redress the meagre progress made in recent decades in defining the molecular basis of pathogenicity of Aspergillus fumigatus 2) Lead us directly to the fungal processes we need to target with new drugs 3) Hugely reduce the number of animal infection studies performed by other scientists on A. fumigatus 4) Help us to optimise high throughput manipulation of A. fumigatus for future drug screens.

由真菌引起的传染病是一个世界性的问题，造成的死亡人数至少与疟疾和肺结核一样多。对于由孢子形成霉菌引起的感染，只有一类可用的口服药物，唑类。临床霉菌分离株中的唑类耐药性正在增加，并且该问题的一个可能原因是唑类在环境中作为农业杀真菌剂的广泛使用。因此，迫切需要开发新的抗真菌治疗方法。真菌颗粒（或孢子），不断存在于我们呼吸的空气中，能够在人类肺部存活。通常，这发生在受感染个体的免疫系统没有发挥最佳作用时。通常，它涉及真菌孢子的萌发（类似于植物种子）和称为菌丝的长真菌细胞的生长。菌丝通过分泌酶穿透并溶解肺组织。我们呼吸的空气中的主要真菌病原体被称为烟曲霉，受癌症影响的人，或需要器官移植的人，特别容易受到致命感染。烟曲霉毒素的存在以及在处理烟曲霉毒素时遇到的技术困难限制了这一领域的进展。我们不完全理解为什么曲霉菌能够在肺环境中存活。我们迫切需要更多关于这种病原体的基本信息，这样我们就可以开始设计未来的治疗方法。当科学家试图弄清楚细胞过程如何工作时，第一个调用的端口通常是转录因子。转录因子同时控制许多基因的活性，因此它们在细胞过程中拥有巨大的力量。如果一个转录因子对一个特定的过程很重要，那么我们就可以看看它调节的是哪些基因，这将为该过程的生物学基础提供线索。如果一个转录因子不参与，那么它和它调节的所有基因都可以从研究中排除。通过询问哪些转录因子控制A.通过研究烟曲霉菌引起疾病的机制，我们可以实现对驱动感染的监管网络的全球视野。我们打算将这种方法与最先进的DNA测序技术结合起来，以获得对A.烟曲霉致病性，小鼠使用最少。我们搜索了A。烟曲霉基因组转录因子基因，并建立了401 A.烟曲霉突变体，每一个都缺乏转录因子。为了了解哪些转录因子对哺乳动物肺部感染很重要，我们将用突变体的集合感染小鼠，看看哪些不能在肺内存活。我们发明了一种方法可以同时测试许多变种人。这包括将所有突变体感染到一只小鼠中，并在处死后，对从肺部提取的DNA进行测序，以告诉我们哪些突变体成功存活。我们将利用这种方法来揭示所有的转录因子驱动致病性（P-TF）在A。烟熏。因此，将使用尽可能少的动物。当我们鉴定了所有的P-TF后，我们将努力鉴定由每个P-TF调节的基因。为了做到这一点，我们将使用两种方法。首先，我们用化学方法将转录因子固定在A上。烟曲霉DNA，以确定每个转录因子的结合位点。其次，我们将研究缺乏P-TF的突变体的基因表达，并找出哪些基因变得失调。这项研究的成功将1）弥补近几十年来在确定烟曲霉致病性的分子基础方面取得的微薄进展2）直接引导我们找到我们需要用新药靶向的真菌过程3）减少其他科学家对A. 4）帮助我们优化A.用于未来的药物筛查。

项目成果

Exploring a novel genomic safe-haven site in the human pathogenic mould Aspergillus fumigatus.

探索人类致病霉菌烟曲霉中的一个新的基因组避风港位点。

• DOI: 10.1016/j.fgb.2022.103702
• 发表时间: 2022
• 期刊: FG & B
• 作者: Furukawa T

On the lineage of Aspergillus fumigatus isolates in common laboratory use.

• DOI: 10.1093/mmy/myaa075
• 发表时间: 2021-01-04
• 期刊: Medical mycology
• 影响因子: 2.9
• 作者: Bertuzzi M;van Rhijn N;Krappmann S;Bowyer P;Bromley MJ;Bignell EM
• 通讯作者: Bignell EM

Refining the pH response in Aspergillus nidulans: a modulatory triad involving PacX, a novel zinc binuclear cluster protein.

• DOI: 10.1111/mmi.13173
• 发表时间: 2015-12
• 期刊: Molecular microbiology
• 影响因子: 3.6
• 作者: Bussink HJ;Bignell EM;Múnera-Huertas T;Lucena-Agell D;Scazzocchio C;Espeso EA;Bertuzzi M;Rudnicka J;Negrete-Urtasun S;Peñas-Parilla MM;Rainbow L;Peñalva MÁ;Arst HN Jr;Tilburn J
• 通讯作者: Tilburn J

Anti-Aspergillus Activities of the Respiratory Epithelium in Health and Disease.

• DOI: 10.3390/jof4010008
• 发表时间: 2018-01-08
• 期刊: Journal of fungi (Basel, Switzerland)
• 作者: Bertuzzi M;Hayes GE;Icheoku UJ;van Rhijn N;Denning DW;Osherov N;Bignell EM
• 通讯作者: Bignell EM

Secondary metabolite arsenal of an opportunistic pathogenic fungus

• DOI: 10.1098/rstb.2016.0023
• 发表时间: 2016-12-05
• 期刊: PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES
• 影响因子: 6.3
• 作者: Bignell, Elaine;Cairns, Timothy C.;Keller, Nancy P.
• 通讯作者: Keller, Nancy P.