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Resistance-gene-guided genome mining for novel antifungal compounds

抗性基因引导的新型抗真菌化合物基因组挖掘

基本信息

批准号：
10368095
负责人：
BERL Ray OAKLEY
金额：
$ 19.14万
依托单位：
UNIVERSITY OF KANSAS LAWRENCE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-08 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10368095
关键词：
Acquired Immunodeficiency Syndrome Africa South of the Sahara Amphotericin B Anabolism Antifungal Agents Area Aspergillus Aspergillus nidulans Asthma Biological Candida albicans Candida auris Catalytic Domain Cell Wall Cessation of life Computers Cryptococcus neoformans Databases Disease Elongation Factor Engineering Enzymes Fungal Genome Fungi Model Gene Cluster Gene Expression Genes Genome Growth Human Immune Infrastructure Institutes Intravenous Joints Learning Logic Lovastatin Malaria Mammals Medical Metabolism Mining Mycoses Natural Resistance Nature Organ Transplantation Organism Pathogenicity Pathway interactions Patients Penicillins Persons Pneumonia Process Production Proteins Renaissance Research Resistance Role Source Structure System Systemic infection Testing Therapeutic Time Translations Tuberculosis Variant base cancer transplantation efficacy evaluation fungus gene product genome sequencing in silico inhibitor metabolome novel novel strategies pathogen pathogenic fungus resistance gene success

项目摘要

PROJECT SUMMARY Approximately 1.5 million people die each year, worldwide, from systemic fungal infections. The high death toll highlights the limitations of current treatments and, in addition, there is a growing problem with resistance to existing antifungals. There is, thus, a compelling need for new and better antifungal agents. Surprisingly, fungi are a good source of antifungals, the echinocandins being a prime example. The antifungals produced by fungi are secondary metabolites (SMs)--compounds that are not essential for viability but confer a selective advantage to the producing organism, often by inhibiting growth of its competitors. The fact that they often inhibit important biological activities has made them a prime source of medically valuable compounds. Fungal SM biosynthetic pathways are encoded by clusters of coordinately regulated genes. Genome projects have revealed that there are vastly more SM gene clusters than known SMs, and, thus, that the greater fungal secondary metabolome (all SMs made by all 1,000,000+ species of fungi) is much larger than was previously thought. To exploit the diversity of fungal SMs efficiently, we need to be able to identify clusters that produce compounds with desired activities. A promising approach to this end, called resistance-gene-directed genome mining, takes advantage of the fact that some SM biosynthetic gene clusters (BGCs) contain a gene that has no role in SM biosynthesis but, rather, encodes a resistant form of the protein inhibited by the compound produced by the cluster. Expression of this gene makes the producing organism resistant to the compound it produces. We have developed an approach, and computer application, that has allowed us to identify SM BGCs in the Joint Genomes Institute database that we predict will produce compounds that inhibit two proteins that are essential for fungal growth. They are FKS1, the catalytic subunit of an enzyme required for fungal cell wall biosynthesis, and YEF3, a translation elongation factor found only in fungi. These proteins are not present in mammals and inhibiting their activities should not be toxic to humans. Because expressing SMs in their native organisms is usually very difficult, we propose to express representative BGCs predicted to produce FKS1 and YEF3 inhibitors in the model fungus Aspergillus nidulans, which we have engineered for heterologous expression of fungal SMs. We propose several approaches, building on our successes in this area, that we will evaluate for efficacy. Heterologously expressed compounds will be purified and their structures will be determined. They will be tested for activity against the pathogenic fungi Candida albicans and Cryptococcus neoformans as well as wild-type A. nidulans which is a useful surrogate for pathogenic species of Aspergillus.

项目概要全球每年约有 150 万人死于全身性真菌感染。死亡人数居高不下强调了当前治疗的局限性，此外，耐药性问题日益严重现有的抗真菌药物。因此，迫切需要新的、更好的抗真菌剂。令人惊讶的是，真菌是抗真菌药物的良好来源，棘白菌素就是一个很好的例子。真菌产生的抗真菌剂是次生代谢物 (SM)——对生存能力不是必需的但具有选择性优势的化合物通常通过抑制其竞争对手的生长来影响生产生物体。事实上，他们经常抑制重要的生物活性使它们成为具有医学价值的化合物的主要来源。真菌SM生物合成通路由协调调节的基因簇编码。基因组计划表明，有 SM 基因簇比已知的 SM 多得多，因此，更大的真菌次级代谢组（所有 SM 由所有 1,000,000 多种真菌产生）比之前想象的要大得多。为了利用为了有效地识别真菌 SM 的多样性，我们需要能够识别产生具有所需化合物的簇活动。为此，一种有前途的方法被称为抗性基因指导的基因组挖掘，它利用了一些 SM 生物合成基因簇 (BGC) 含有在 SM 生物合成中没有作用的基因相反，它编码被簇产生的化合物抑制的蛋白质的抗性形式。该基因的表达使生产生物体对其产生的化合物产生抗性。我们有开发了一种方法和计算机应用程序，使我们能够识别联合作战中的 SM BGC 我们预测基因组研究所数据库将产生抑制两种必需蛋白质的化合物用于真菌生长。它们是 FKS1，真菌细胞壁生物合成所需的酶的催化亚基， YEF3，一种仅在真菌中发现的翻译延伸因子。这些蛋白质不存在于哺乳动物中抑制它们的活动不应对人类有毒。因为在其天然生物体中表达 SM 通常非常困难，我们建议表达预测产生 FKS1 和 YEF3 抑制剂的代表性 BGC 在模型真菌构巢曲霉中，我们对其进行了改造，用于真菌 SM 的异源表达。基于我们在这一领域取得的成功，我们提出了几种方法，我们将评估这些方法的有效性。异源表达的化合物将被纯化并确定其结构。他们将接受考验针对致病性真菌白色念珠菌、新型隐球菌以及野生型 A 的活性。构巢菌，它是曲霉菌致病菌的有用替代品。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Manipulation of the Global Regulator mcrA Upregulates Secondary Metabolite Production in Aspergillus wentii Using CRISPR-Cas9 with In Vitro Assembled Ribonucleoproteins.

DOI：
10.1021/acschembio.2c00456
发表时间：
2022-10-21
期刊：
ACS CHEMICAL BIOLOGY
影响因子：
4
作者：
Yuan, Bo;Keller, Nancy P.;Oakley, Berl R.;Stajich, Jason E.;Wang, Clay C. C.
通讯作者：
Wang, Clay C. C.

Conversion of Polyethylenes into Fungal Secondary Metabolites.

DOI：
10.1002/anie.202214609
发表时间：
2023-01-23
期刊：
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
影响因子：
16.6
作者：
Rabot, Chris;Chen, Yuhao;Bijlani, Swati;Chiang, Yi-Ming;Oakley, C. Elizabeth;Oakley, Berl R.;Williams, Travis J.;Wang, Clay C. C.
通讯作者：
Wang, Clay C. C.

Computer-aided, resistance gene-guided genome mining for proteasome and HMG-CoA reductase inhibitors.

DOI：
10.1093/jimb/kuad045
发表时间：
2023-02-17
期刊：
Journal of industrial microbiology & biotechnology
影响因子：
3.4
作者：
通讯作者：

Identification of the chaA and fwA Spore Color Genes of Aspergillus nidulans.

构巢曲霉 chaA 和 fwA 孢子颜色基因的鉴定。

DOI：
10.3390/jof10020104
发表时间：
2024
期刊：
Journal of fungi (Basel, Switzerland)
影响因子：
0
作者：
Oakley,CElizabeth;BartonJr,ThomasS;Oakley,BerlR
通讯作者：
Oakley,BerlR

A heterologous expression platform in Aspergillus nidulans for the elucidation of cryptic secondary metabolism biosynthetic gene clusters: discovery of the Aspergillus fumigatus sartorypyrone biosynthetic pathway.

DOI：
10.1039/d3sc02226a
发表时间：
2023-10-18
期刊：
CHEMICAL SCIENCE
影响因子：
8.4
作者：
Lin, Shu-Yi;Oakley, C. Elizabeth;Jenkinson, Cory B.;Chiang, Yi-Ming;Lee, Ching-Kuo;Jones, Christopher G.;Seidler, Paul M.;Nelson, Hosea M.;Todd, Richard B.;Wang, Clay C. C.;Oakley, Berl R.
通讯作者：
Oakley, Berl R.