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: 10689662
• 负责人: Paul Stephen Keim
• 金额: $ 154.27万
• 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/10689662
• 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.

整体部分 标题：早期体内表达的抗原及其在毒力、免疫反应和疫苗中的作用 球孢子菌病。 概括 球孢子菌病，又称谷热病（VF），是由两种不同的真菌引起的重要真菌病。 球孢子菌属物种，导致区域性重要死亡率甚至更高发病率。我们有 组建了一个团队来定义人类和动物对 VF 的免疫反应发生的变化，并使用 这些知识可用于设计新疫苗和诊断测试。这项工作将利用我们详细的 感染早期阶段球孢子菌基因表达模式的观察。我们假设 一些球孢子菌早期基因是毒力因子，对于引起疾病至关重要。研究项目1 将通过使用 CRISPR-Cas9 技术的基因敲除和蜡虫毒力测试来测试其作用 (Galleria) 和小鼠 VF 模型。关键毒力因子将成为诊断和疫苗目标。此外 针对蜡虫和小鼠模型，我们开发了一种非人类灵长类动物（猪尾猕猴）， 与人类的 VF 非常相似。在人类和脊椎动物模型中，T 细胞的作用不能被 研究项目 2 将使用重点深度 DNA 测序来识别 T 细胞类别 响应早期表达的球孢子菌基因而发育的受体（TCR）。我们将生成TCR 来自跨越病原体流行区的三个临床地点的患者的序列。 TCR 来自患者的库将用于识别新的诊断特征（例如公共 TCR），并且还可以帮助 确定对可作为疫苗开发目标的关键抗原的免疫反应。因此，TCR 和 早期毒力基因代表了疫苗设计的优秀候选基因，将在研究中进行探索 项目 3 使用基于核酸 (NA) 的疫苗（RNA 和 DNA），可以快速测试大量抗原 通过对小鼠进行免疫来抵抗感染。 DNA 疫苗将基于黄金交付 纳米颗粒和基因枪，而 mRNA 采用自我复制 RNA 分子 (repRNA) 和脂质 无机纳米颗粒（LION）。两者都是经过验证的技术，正在进入其他领域的临床试验。 疾病。我们在 NA 疫苗小鼠研究中的目标是确定最佳抗原和递送方式 在 NHP 模型中进行疫苗测试并确定其免疫保护机制。这部作品仅 通过七个不同机构（包括三个临床中心）的研究人员的综合努力， 因为没有任何一个机构具备必要的专业知识和基础设施。虽然我们将生成 关于球孢子菌和 VF 的基础知识，我们还将在以下方面取得转化进展： 预防和诊断疾病。