Early in vivo expressed antigens and their role in virulence, immune response, and vaccines for coccidioidomycosis

早期体内表达的抗原及其在球孢子菌病毒力、免疫反应和疫苗中的作用

• 批准号: 10356628
• 负责人: Bridget Marie Barker
• 金额: $ 26.32万
• 依托单位: NORTHERN ARIZONA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-24 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10356628
• 关键词: Adjuvant; Algorithms; Animal Model; Animals; Antibodies; Antigens; Attenuated; Bioinformatics; Biology; CRISPR/Cas technology; Candidate Disease Gene; Cellular Immunity; Centers for Disease Control and Prevention (U.S.); Coccidioides; Coccidioides immitis; Coccidioides posadasii; Coccidioidomycosis; Collaborations; Data; Data Analyses; Development; Diagnostic; Diagnostic tests; Disease; Disease Outcome; Foundations; Fungal Drug Resistance; Gene Deletion; Gene Expression Profiling; Gene Proteins; Gene Silencing; Gene Targeting; Generations; Genes; Genetic Transcription; Genomics; Goals; Growth; Health; Human; Immune; Immune response; Immunity; Immunization; In Vitro; Incidence; Infection; Investigation; Knock-out; Knowledge; Length of Stay; Life; Lung; Methods; Mus; Mycoses; Nucleic Acid Vaccines; PAWR protein; Parents; Pathogenesis; Pathogenicity; Patients; Play; Population; Production; Proteins; Reaction; Recombinant Proteins; Recombinants; Reporting; Research; Research Project Grants; Research Proposals; Role; Safety; Serology test; Sister; Structure; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Epitopes; Testing; Therapeutic; Vaccination; Vaccine Antigen; Vaccine Design; Vaccines; Virulence; Virulence Factors; Vision; adaptive immune response; base; cost; desert fever; diagnostic tool; disorder prevention; gene function; genetic analysis; genetic approach; genome analysis; genome sequencing; genomic data; immunogenic; immunogenicity; improved; in silico; in vivo; nano-string; novel; pathogen; pathogenic fungus; protein complex; response; reverse genetics; side effect; targeted biomarker; tool; transcriptome sequencing; treatment strategy; vaccine candidate; vaccine development; whole genome

Project Summary/Abstract (Research Project 1) Emerging and endemic mycoses have a growing impact on human health on a global scale and little is known about virulence mechanisms for most emerging fungal pathogens. Coccidioides immitis and C. posadasii cause coccidioidomycosis (aka Valley Fever), which is of specific concern due to recent rapid increases in US disease incidence with a more than a 10-fold increase of reports over the last 20 years. The Centers for Dis- ease Control and Prevention (CDC) report that the average cost for treating Valley fever is $50,000 per patient, including extensive hospital stays and treatment regimes. Advanced genomic and genetic analyses of these fungal pathogens can help improve our understanding of mechanisms of pathogenicity, support the identifica- tion of new biomarker targets, and represent the foundational framework that is necessary for development of diagnostic tools, treatment strategies and prevention of disease via vaccination. Currently, few specific viru- lence mechanisms have been investigated in Coccidioides. Complicating these investigations is the lack of in vivo data, as the biology and gene expression analysis of in vitro cultures does not replicate the infective se- quence. We have remedied this gap by detailed transcriptional analysis of both Coccidioides species in mouse lungs five days after infection. Based on our transcriptional analyses and existing genomic data, plus in silico functional predictions, we have identified 26 putative antigens/virulence factors. These Coccidioides spp. tran- scripts are highly expressed under in vivo conditions and are predicted to be small secreted proteins. Our working hypothesis is that these proteins will be antigenic and potentially virulence factors. We will test this hy- pothesis by creating CRISPR-Cas9 gene deletion strains. These strains will be tested for virulence in two ani- mal models in collaboration with the Animal Core. We will further investigate these genes’ functions in both Coccidioides species, specifically the effects on development of the parasitic structure (spherule), virulence, and host and pathogen response in vivo using nanoString. RNAseq analysis of gene deletion strains compared to parent strain will be used to compare transcriptional networks. Analysis of these data will provide new in- sights into gene functions, and effectively double the number of gene deletion strains investigated for this path- ogen, which will increase our understanding of fungal pathogenesis mechanisms. These data will be used by the other two Research Projects to identify T cell epitopes and the cognate T cell receptors for understanding the host immune response and possible advanced diagnostic tests, as well as to identify targets for nucleic acid vaccines. Programming the immune response toward early infection virulence factors has the great poten- tial to provide protective immunity to Valley Fever. In total, this proposal represents a unique pairing of ad- vanced genomic data analysis and functional assessments in an important, yet understudied, fungal pathogen.

项目概要/摘要（研究项目1） 新发和地方性真菌病在全球范围内对人类健康的影响越来越大，但人们知之甚少 关于大多数新兴真菌病原体的毒力机制。 引起球孢子菌病（又名谷热），由于最近在美国的快速增长而引起特别关注 过去 20 年，疾病发病率报告增加了 10 倍以上。 疾病预防控制中心 (CDC) 报告称，治疗山谷热的平均费用为每位患者 50,000 美元， 包括广泛的住院时间和治疗方案的高级基因组和遗传分析。 真菌病原体可以帮助提高我们对致病机制的理解，支持识别 新的生物标志物目标的重刑，并代表了发展所必需的基础框架 目前，很少有特定的病毒可以通过疫苗接种来预防疾病。 球孢子菌的作用机制已被研究，但这些研究的复杂性是缺乏。 体内数据，因为体外培养物的生物学和基因表达分析不能复制感染性序列 我们通过对小鼠中两种球孢子菌的详细转录分析弥补了这一空白。 根据我们的转录分析和现有的基因组数据，以及计算机模拟，感染后五天的肺部。 通过功能预测，我们已经鉴定出 26 种假定的抗原/毒力因子。 脚本在体内条件下高度表达，预计是小分泌蛋白。 工作假设是这些蛋白质将是抗原和潜在的毒力因子，我们将测试这种hy-。 通过创建 CRISPR-Cas9 基因缺失菌株的假设，这些菌株将在两种动物中进行毒力测试。 我们将与 Animal Core 合作进一步研究这些基因在 mal 模型中的功能。 球孢子菌属物种，特别是对寄生结构（小球）发育、毒力、 并使用nanoString RNAseq分析基因缺失菌株的体内宿主和病原体反应进行比较。 与亲本菌株的比较将用于比较转录网络。对这些数据的分析将提供新的信息。 着眼于基因功能，并有效地将为此途径研究的基因缺失菌株的数量加倍- ogen，这将增加我们对真菌发病机制的理解。 另外两个研究项目旨在识别 T 细胞表位和同源 T 细胞受体以供理解 宿主免疫反应和可能的高级诊断测试，以及确定核酸靶点 酸性疫苗对早期感染毒力因子的免疫反应具有巨大的潜力。 总体而言，该提案代表了一种独特的广告组合。 对一种重要但尚未得到充分研究的真菌病原体进行先进的基因组数据分析和功能评估。