Coccidioidomycosis:Genome Comparison, Selection and Transcription.

描述（由申请人提供）：我们的长期目标是开发预防和治疗球孢子菌病的方法，这是一种由真菌粗球孢子菌和波氏球孢子菌引起的健康人的全身性真菌疾病。合作研究人员是约翰泰勒和加里科尔，实验室主任，研究球孢子菌分子发育和进化生物学，分别。我们的目标是在这个建议是集中的比较基因组学和球孢子菌的基因表达的工具，以确定重要的致病性和毒力的基因。我们的两个发现支持了我们的建议：1）已知对球孢子菌致病性重要的基因的破坏降低了其在哺乳动物中的毒力，2）球孢子菌在五个地理种群中显示出遗传变异。每个/球孢子菌属物种的完整注释基因组及其非致病性亲属Uncinocarpus reesii的基因组的可用性是支持我们项目的主要支柱。我们可以通过在基因组中寻找致病基因，同时考虑自然变异，大大增加治疗或预防的靶点数量。我们将通过小鼠毒力试验来评估我们对这些基因的假设。我们的具体目标是：鉴定与Uncinocarpus相比球孢子菌属特有的基因，鉴定在球孢子菌属物种之间和与Uncinocarpus相比显示阳性选择的基因，以及鉴定在体外和体内在腐殖和寄生阶段之间具有显著转录差异的球孢子菌属基因。从这一处于正选择下并在寄生阶段上调的独特基因的干部中，我们将鉴定一组推定的致病性基因，我们将通过评估野生型菌株、其中目标基因已被敲除的WT菌株和目标基因被替换的KO菌株在小鼠中的毒力来测试这些基因。通过现有的合作，我们的实验室已经研究并发表了关于球孢子菌致病基因的自然选择，并评估了在腐殖和寄生阶段的转录。与公共卫生的相关性：近10%的美国人生活在球孢子菌病流行的地区，大约5%的感染者发展为危及生命的疾病。症状感染赋予长期免疫力，因此接种疫苗是可能的。同样地，当致病性基因被禁用时，毒力降低，这表明这些基因产物的药物靶向应该是有效的。

DESCRIPTION (provided by applicant): Our long-term objective is to develop methods to prevent and treat coccidioidomycosis, a systemic fungal disease of otherwise healthy humans caused by the fungi Coccidioides immitis and Coccidioides posadasii. The collaborating investigators are John Taylor and Garry Cole, directors of laboratories that study Coccidioides molecular developmental and evolutionary biology, respectively. Our goal in this proposal is to focus the tools of comparative genomics and gene expression on Coccidioides to identify genes important to pathogenicity and virulence. Two of our findings underlie our proposal: 1) disruption of genes known to be important to the pathogenicity of Coccidioides reduces its virulence in mammals, and 2) Coccidioides shows genetic variation among five geographic populations. The availability of complete, annotated genomes of each /coccidioides species and the genome of their non-pathogenic relative, Uncinocarpus reesii, is a major pillar of support for our project. We can greatly increase the number of targets for therapy or prevention by searching the genome for pathogenicity genes, while accounting for natural variation. We will evaluate our hypotheses about these genes by virulence tests in mice. Our specific aims are to: identify genes unique to Coccidioides as compared to Uncinocarpus, identify genes that show positive selection between Coccidioides species and compared to Uncinocarpus, and identify genes in Coccidioides with significant transcription differences between the saprobic and parasitic phases, both in vitro and in vivo. From this cadre of unique genes which are under positive selection and upregulated during the parasitic phase, we will identify a pool of putative pathogenicity genes that can we will test by assessing virulence in mice of the wild-type strain, the WT strain in which the gene of interest has been knocked out, and the KO strain with the gene of interest replaced. Via existing collaboration, our laboratories have studied and published on natural selection of Coccidioides pathogenicity genes and assessed transcription in the saprobic and parasitic phases. Relevance to public health: Nearly 10% of Americans live in areas endemic to coccidioidomycosis, and approximately 5% of those infected develop life-threatening disease. Symptomatic infections confer long term immunity, so vaccination is possible. Likewise, virulence is reduced when pathogenicity genes are disabled, which indicates that pharmaceutical targeting of these gene products should be efficacious.