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: 10891793
• 负责人: Paul Stephen Keim
• 金额: $ 139.02万
• 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/10891793
• 关键词: Animal Model; Animals; Antibodies; Antigens; Basic Science; CRISPR/Cas technology; Clinical; Clinical Trials; Coccidioides; Coccidioides immitis; Coccidioides posadasii; Coccidioidomycosis; Communities; Complex; DNA; DNA Vaccines; DNA sequencing; Data; Development; Diagnostic; Diagnostic tests; Disease; Epitopes; Gene Expression; Gene Expression Profile; Genes; Goals; Human; Immune; Immune response; Immunization; Infection; Infrastructure; Insecta; Institution; Knock-out; Knowledge; Link; Lipids; Location; Macaca nemestrina; Messenger RNA; Modality; Modeling; Morbidity - disease rate; Mus; Nucleic Acid Amplification Tests; Nucleic Acid Vaccines; Nucleic Acids; Pathogenesis; Patients; Pattern; Peptides; Population; Prevention; Process; Protein Secretion; Proteins; RNA; RNA replication; RNA vaccination; RNA vaccine; Research; Research Personnel; Research Project Grants; Role; Serology test; T cell response; T-Cell Receptor; T-Lymphocyte; T-cell receptor repertoire; Technology; Testing; Therapeutic; Therapeutic Agents; Time; Translational Research; United States; Vaccination; Vaccine Antigen; Vaccine Design; Vaccines; Vertebrates; Virulence; Virulence Factors; Waxes; Work; clinical research site; companion diagnostics; desert fever; design; diagnostic signature; diagnostic strategy; diagnostic tool; disease diagnosis; gene gun; gene product; human disease; in vivo; insight; knockout gene; longitudinal analysis; mortality; mouse model; nano-string; nanoGold; nanoparticle; nonhuman primate; novel; novel diagnostics; novel vaccines; pathogen; prevent; respiratory; response; screening; serological marker; therapeutic evaluation; tool; transcriptome sequencing; vaccine candidate; vaccine development; vaccine evaluation; virulence gene

OVERALL Section Title: Early in vivo expressed antigens and their role in virulence, immune response, and vaccines for coccidioidomycosis. SUMMARY Coccidioidomycosis, also known as Valley Fever (VF), is an important fungal disease caused by two different Coccidioides species that results in regionally important mortality and even greater morbidity. We have assembled a team to define the changes that occur in human and animal immune responses to VF, and to use this knowledge for designing new vaccines and diagnostic tests. This work will capitalize upon our detailed observations of Coccidioides gene expression patterns during the earliest stages of infections. We hypothesize that some Coccidioides early genes are virulence factors and critical for causing disease. Research Project 1 will test their role through gene knockouts using CRISPR-Cas9 technology and virulence testing in wax worm (Galleria) and mouse VF models. Critical virulence factors will become diagnostic and vaccine targets. In addition to the wax worm and mouse models, we develop a non-human primate (pig-tailed macaques) that will more closely resemble VF in humans. In humans and vertebrate animal models, the role of T cells cannot be overemphasized and Research Project 2 will use focused deep DNA sequencing to identify classes of T cell receptors (TCR) that develop in response to early expressed Coccidioides genes. We will generate TCR sequences from patients at three clinical locations that span the endemic zones for the pathogen. The TCR repertoire from patients will be used to identify novel diagnostic signatures (e.g., public TCRs) and, also, help identify immune responses to key antigens that can be targeted for vaccine development. Hence, both TCR and early virulence genes represent excellent candidates for vaccine design that will be explored in Research Project 3 using nucleic acid (NA) based vaccines (RNA and DNA) that can rapidly test a large panel of antigens through the immunization of mice against infection. The DNA vaccine will be based upon delivery on gold nanoparticles and Gene Gun, while the mRNA employs self-replicating RNA molecules (repRNA) and a Lipid InOrganic Nanoparticle (LION). Both are proven technologies that are moving forward into clinical trials for other diseases. Our goal in the NA vaccine mouse studies is to identify the best antigens and delivery modality for vaccine testing in the NHP model and to define their immune mechanisms of protection. This work is only possible through the integrated efforts of investigators at seven different institutions, including three clinical sites, as no single institution has the requisite breadth of expertise and infrastructure. While we will generate fundamental knowledge about Coccidioides and VF, we will also make translational advances towards preventing and diagnosing the disease.

整个部分 标题：体内早期表达抗原及其在毒力，免疫反应和疫苗中的作用 球菌病。 概括 球虫菌病，也称为山谷发烧（VF），是由两种不同的真菌疾病 球虫下导致区域重要的死亡率甚至更大的发病率。我们有 组装一个团队来定义人类和动物免疫反应中发生的变化，并使用 这些知识用于设计新的疫苗和诊断测试。这项工作将利用我们的详细 在最早的感染阶段观察到球球菌基因表达模式。我们假设 某些球虫的早期基因是毒力因素，对于引起疾病至关重要。研究项目1 将使用CRISPR-CAS9技术和蜡蠕虫中的毒力测试通过基因敲除测试其角色 （Galleria）和鼠标VF模型。关键的毒力因子将成为诊断和疫苗靶标。此外 对于蜡蠕虫和鼠标型号，我们开发了一种非人类灵长类动物（猪尾猕猴），它将更多 在人类中非常类似于VF。在人类和脊椎动物模型中，T细胞的作用不能 过度强调，研究项目2将使用重点的深度DNA测序来识别T细胞类别 响应早期表达的球虫基因而发展的受体（TCR）。我们将生成TCR 来自病原体的特有区域的三个临床部位的患者的序列。 TCR 患者的曲目将用于识别新颖的诊断特征（例如，公共TCR），还可以帮助您提供帮助。 确定可以针对疫苗开发的关键抗原的免疫反应。因此，TCR和 早期毒力基因代表了将在研究中探索的疫苗设计的出色候选者 使用基于核酸（NA）的疫苗（RNA和DNA）的项目3，该疫苗可以快速测试一大批抗原 通过小鼠免疫感染。 DNA疫苗将基于黄金的递送 纳米颗粒和基因枪，而mRNA采用自我复制的RNA分子（retra）和脂质 无机纳米颗粒（狮子）。两者都是经过验证的技术，这些技术正在进行其他方面的临床试验 疾病。我们在NA疫苗小鼠研究中的目标是确定最佳的抗原和递送方式 NHP模型中的疫苗测试并定义其免疫机制。这项工作只是 通过研究人员在七个不同机构的综合努力，包括三个临床地点， 由于没有一个机构具有必要的专业知识和基础设施。虽然我们会生成 关于球虫剂和VF的基本知识，我们还将转化发展 防止和诊断疾病。