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.

项目摘要/摘要（研究项目1） 在全球范围内对人类健康的影响越来越大，鲜为人知 关于大多数新兴真菌病原体的病毒机制。球球菌Immitis和C. posadasii 导致球虫病（又称山谷热），这是由于最近迅速增加而引起的特别关注 在过去20年中，疾病事件的报告增加了10倍以上。 dist的中心 Ease Control and Fecention（CDC）报告说，治疗山谷发烧的平均成本为每位患者50,000美元， 包括广泛住院和治疗方案。这些的晚期基因组和遗传分析 真菌病原体可以帮助我们提高我们对致病性机制的理解，支持鉴定 - 新的生物标志物目标的特征，代表开发的基础框架 诊断工具，治疗策略和通过疫苗预防疾病。目前，很少有特定的viru- 在球虫毒剂中已经研究了透镜机制。使这些调查复杂化的是缺乏 体内数据，因为体外培养物的生物学和基因表达分析并未复制感染性疾病 quence。我们已经通过对小鼠中两个球虫种类的详细转录分析来修复了这一差距 感染五天后肺。基于我们的转录分析和现有基因组数据，以及计算机 功能预测，我们已经确定了26种推定的抗原/病毒因子。这些球虫属属。 tr 脚本在体内条件下高度表达，预计为小的分泌蛋白质。我们的 工作假设是这些蛋白质将是抗原和潜在的病毒因素。我们将测试这个问题 通过创建CRISPR-CAS9基因删除菌株来进行放大。这些菌株将在两个芳族中测试病毒 MAL模型与动物核心合作。我们将进一步研究这些基因的功能 球虫种类，特别是对寄生结构（Spherule），病毒，，病毒的影响 以及使用纳米串在体内的宿主和病原体反应。基因缺失菌株的RNASEQ分析 到父菌株将用于比较转录网络。对这些数据的分析将提供新的信息 瞄准器的视力为基因功能，并有效地将基因缺失菌株的数量翻了一番。 Gen，这将增加我们对真菌发病机制的理解。这些数据将由 其他两个研究项目以识别T细胞表位和同源T细胞受体以理解 宿主免疫反应和可能的晚期诊断测试，并确定核的目标 酸性疫苗。编程对早期感染病毒因素的免疫反应具有很大的潜在 tial可为山谷发烧提供受保护的免疫力。总的来说，该提议代表了aD的独特配对 在重要但被理解的真菌病原体中，基因组数据分析和功能评估。