A molecular toolbox for hypothesis testing in the dermatophytes

用于皮肤癣菌假设检验的分子工具箱

• 批准号: 7763172
• 负责人: Theodore C. White
• 金额: $ 28.63万
• 依托单位: SEATTLE BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-03 至 2011-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7763172
• 关键词: Address; Animals; Area; Arthrodermataceae; Aspergillus; Candida; Cells; Child; Childhood; Complex; Cryptococcus; DNA; Depressed mood; Detection; Development; Diagnosis; Diagnostic; Exhibits; Expressed Sequence Tags; Funding; Gene Deletion; Gene Expression; Genotype; Goals; Growth; Head; Healthcare Systems; Human; Individual; Infection; Infection prevention; Institutes; Larva; Methods; Microarray Analysis; Modeling; Molds; Molecular; Moths; Mutagenesis; Mycoses; Nail plate; Nonhomologous DNA End Joining; Pain; Pathogenicity; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Phenotype; Population; Prevention; Prevention strategy; Prophylactic treatment; Research; Research Personnel; Resistance; Rhizobium radiobacter; Rodent; Route; Scalp structure; Skin; Study models; Surface; Symptoms; Techniques; Testing; Tinea; Tinea Capitis; Tinea Pedis; Trichophyton; United States; Vaccines; Virulence; Waxes; cDNA Arrays; economic impact; fungus; genome sequencing; homologous recombination; improved; keratinocyte; novel; overexpression; pathogen; public health relevance; theories; tool

DESCRIPTION (provided by applicant): Dermatophytes are molds, closely related to Aspergillus and the dimorphic fungi. These molds cause infections within the skin and nails, including athlete's foot, the most common fungal infection worldwide. In the United States, the economic impact of athlete's foot alone on the health care system is estimated to exceed $400 million a year. Similarly, fungal infection of the head is a frequent pediatric infection in major urban centers in the U.S. and worldwide, and is especially common in socio-economically depressed populations. In a recent study, 22-50% of children in a U.S. daycare exhibited symptoms of dermatophytic scalp infection. The dermatophytes remain under-studied and under-funded. Recent genome sequencing and EST analysis of the dermatophytes will enable researchers to generate hypotheses concerning the dermatophytes with the ultimate goal to develop methods for the detection (diagnostics), treatment (drugs), and prevention (vaccines or prophylaxis) of dermatophyte infections. A standard method for testing hypotheses is to alter the genotype of the fungal cell and then assess the resulting phenotype. However, the tools for altering the genotype and assessing the phenotype are severely limited for dermatophytes. The objective of this proposal is to develop a molecular toolbox that will allow researchers to test hypotheses relating to dermatophyte pathogenicity. The specific aims of the proposal are as follows: Aim 1 - To develop techniques for altering the dermatophyte genotype, using split markers to maximize homologous recombination, and Agrobacterium tumefaciens transformation for random mutagenesis and gene expression control. Aim 2 - To evaluate alternate host pathogen models for assessing dermatophyte phenotypes. Human skin "rafts" will be used to study the interactions of the fungus with differentiated keratinocytes. Dermatophyte infections will also be tested in Galleria mellonella, a wax moth larvae which is currently in use for other fungal infections. When completed, these aims will provide hypothesis-testing tools that are essential, in combination with genome sequences and expression analysis, to address questions of pathogenicity and virulence, with the ultimate goal of the development of diagnostics, treatments and prevention strategies for the dermatophytes. PUBLIC HEALTH RELEVANCE: Dermatophytes are pathogenic fungi responsible for a variety of skin and nail infections, including athlete's foot and tinea capitis, a common fungal infection of the head in patients in socio- economically depressed areas. It is estimated that dermatophytes are the most common fungal infection worldwide. This proposal will develop a set of molecular tools that can be used in combination with the soon-to-be-completed genome sequences to address important questions about dermatophyte prevention, treatment and diagnosis.

描述(申请人提供)：皮肤真菌是霉菌，与曲霉菌和二相性真菌关系密切。这些霉菌会导致皮肤和指甲内的感染，包括全球最常见的真菌感染--脚气。在美国，仅脚气对医疗保健系统的经济影响估计就超过每年4亿美元。同样，在美国和世界各地的主要城市中心，头部真菌感染是一种常见的儿科感染，在社会经济低迷的人群中尤其常见。在最近的一项研究中，美国一家托儿所的22%-50%的儿童表现出皮肤真菌头皮感染的症状。对皮肤真菌的研究仍然不足，资金也不足。最近对皮肤真菌的基因组测序和EST分析将使研究人员能够产生关于皮肤真菌的假设，最终目标是开发皮肤真菌感染的检测(诊断)、治疗(药物)和预防(疫苗或预防)的方法。检验假设的标准方法是改变真菌细胞的基因型，然后评估由此产生的表型。然而，改变基因和评估表型的工具对于皮肤真菌来说是非常有限的。这项提议的目标是开发一个分子工具箱，使研究人员能够测试与皮肤真菌致病性有关的假说。该提案的具体目的如下：目标1-开发改变皮肤真菌基因型的技术，使用分裂标记最大限度地进行同源重组，并利用农杆菌转化进行随机突变和基因表达调控。目的2-评价皮肤癣菌表型评估的交替寄主病原体模型。人类皮肤“筏”将被用来研究这种真菌与分化的角质形成细胞的相互作用。皮肤真菌感染也将在Galeria mellonella中进行测试，这是一种蜡蛾幼虫，目前正用于其他真菌感染。完成后，这些目标将提供与基因组序列和表达分析相结合的基本假设检验工具，以解决致病性和毒性问题，最终目标是开发皮肤霉菌的诊断、治疗和预防战略。公共卫生相关性：皮肤真菌是导致各种皮肤和指甲感染的致病真菌，包括足癣和头癣，这是社会经济不景气地区患者头部的一种常见真菌感染。据估计，皮肤癣菌是世界上最常见的真菌感染。这项提议将开发一套分子工具，可以与即将完成的基因组序列结合使用，以解决有关皮肤真菌病预防、治疗和诊断的重要问题。

项目成果

Sequenced dermatophyte strains: growth rate, conidiation, drug susceptibilities, and virulence in an invertebrate model.

• DOI: 10.1016/j.fgb.2010.11.010
• 发表时间: 2011-03
• 期刊: FUNGAL GENETICS AND BIOLOGY
• 影响因子: 3
• 作者: Achterman, Rebecca R.;Smith, Adam R.;Oliver, Brian G.;White, Theodore C.
• 通讯作者: White, Theodore C.

Dermatophyte virulence factors: identifying and analyzing genes that may contribute to chronic or acute skin infections.

• DOI: 10.1155/2012/358305
• 发表时间: 2012
• 期刊: International journal of microbiology
• 影响因子: 3.4
• 作者: Achterman RR;White TC
• 通讯作者: White TC

Dermatophytes activate skin keratinocytes via mitogen-activated protein kinase signaling and induce immune responses.

• DOI: 10.1128/iai.02776-14
• 发表时间: 2015-04
• 期刊: Infection and immunity
• 影响因子: 3.1
• 作者: Achterman RR;Moyes DL;Thavaraj S;Smith AR;Blair KM;White TC;Naglik JR
• 通讯作者: Naglik JR

Discovery of cryptic polyketide metabolites from dermatophytes using heterologous expression in Aspergillus nidulans.

• DOI: 10.1021/sb400048b
• 发表时间: 2013-11-15
• 期刊: ACS SYNTHETIC BIOLOGY
• 影响因子: 4.7
• 作者: Yin, Wen-Bing;Chooi, Yit Heng;Smith, Adam R.;Cacho, Ralph A.;Hu, Youcai;White, Theodore C.;Tang, Yi
• 通讯作者: Tang, Yi