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（由申请人提供）：皮肤植物是霉菌，与曲霉和二态真菌密切相关。这些霉菌引起皮肤和指甲内的感染，包括运动员脚，这是全球最常见的真菌感染。在美国，仅运动员脚对医疗保健系统的经济影响估计每年超过4亿美元。同样，头部的真菌感染是美国和全球主要城市中心的经常感染，在社会经济抑郁症中尤为常见。在最近的一项研究中，美国日托中有22-50％的儿童表现出皮肤哲学头皮感染的症状。皮肤植物仍然研究不足和资金不足。对皮肤植物的最新基因组测序和EST分析将使研究人员能够以最终目标产生有关皮肤植物的假设，以开发检测方法（诊断），治疗（药物）和预防（预防疫苗或预防）（疫苗或预防性）皮肤病感染。测试假设的标准方法是改变真菌细胞的基因型，然后评估所得的表型。但是，改变基因型和评估表型的工具受到皮肤植物的严重限制。该提案的目的是开发一个分子工具箱，该工具箱将使研究人员能够检验与皮肤植物致病性有关的假设。该提案的具体目的如下：目标1-开发用于改变皮肤植物基因型的技术，使用拆分标记来最大程度地提高同源重组，以及tumefaciens农杆菌的随机诱变和基因表达对照。目标2-评估用于评估皮肤植物表型的替代宿主病原体模型。人皮肤“筏”将用于研究真菌与分化角质形成细胞的相互作用。皮肤植物感染也将在Mellonella Galleria Mellonella（一种蜡蛾幼虫）中进行测试，目前正在用于其他真菌感染。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假设测试工具，以解决致病性和毒力问题，以及开发诊断，治疗和预防策略的最终目标。公共卫生相关性：皮肤植物是致病性真菌，负责各种皮肤和指甲感染，包括运动员的脚和Tinea Capitis，这是社会经济抑郁症地区患者的头部常见真菌感染。据估计，皮肤植物是全球最常见的真菌感染。该建议将开发一组分子工具，这些工具可以与即将完成的基因组序列结合使用，以解决有关预防皮肤植物的重要问题。

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