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，球孢子菌病是一种由球孢子菌和波萨达球孢子菌引起的健康人的全身性真菌疾病。合作研究者是约翰·泰勒和加里·科尔，他们分别是研究球虫分子发育和进化生物学的实验室主任。我们的目标是将比较基因组学和基因表达的工具集中在球虫身上，以确定对致病性和毒力重要的基因。我们的两个发现支持了我们的建议：1)已知对球虫致病性重要的基因的破坏降低了它在哺乳动物中的毒力，2)球虫在五个地理种群中表现出遗传变异。获得每个球虫物种的完整、带注释的基因组及其非致病性亲缘种Uncinocarpus reesii的基因组是支持我们项目的主要支柱。在考虑自然变异的同时，通过在基因组中搜索致病性基因，我们可以大大增加治疗或预防目标的数量。我们将通过小鼠毒力测试来评估我们关于这些基因的假设。我们的具体目标是：鉴定与Uncinocarpus相比，coccidiides特有的基因，鉴定与Uncinocarpus相比，coccidiides物种之间表现出正选择的基因，以及鉴定在体外和体内，coccidiides中具有显着转录差异的基因。从这些在寄生阶段处于正选择和上调的独特基因中，我们将确定一个假定的致病基因库，我们将通过评估野生型菌株、敲除感兴趣基因的WT菌株和替换感兴趣基因的KO菌株的小鼠毒力来测试这些基因库。通过现有的合作，我们的实验室已经研究并发表了球虫致病性基因的自然选择，并评估了球虫侵染期和寄生期的转录。与公共卫生相关：近10%的美国人生活在球虫病流行的地区，其中约5%的感染者发展为危及生命的疾病。有症状的感染会产生长期免疫力，因此接种疫苗是可能的。同样，当致病性基因失活时，毒力会降低，这表明药物靶向这些基因产物应该是有效的。

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.