Host Responses to Coccidioides by Human Airway Epithelium

人体气道上皮对球孢子菌的宿主反应

• 批准号: 10616716
• 负责人: Jatin M Vyas
• 金额: $ 21万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-02 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10616716
• 关键词: Address; Animals; Antifungal Antibiotics; Apical; Architecture; Attenuated; CRISPR screen; Cell Communication; Cell Culture Techniques; Cell Line; Cell secretion; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Coccidioides; Coccidioides immitis; Coccidioides posadasii; Complication; Critical Pathways; Data; Direct Costs; Disease; Dissection; Distal; Environment; Epithelial Cells; Epithelium; Extravasation; Foundations; Future; Gene Expression Profile; Genetic Transcription; Host Defense; Human; Human body; IL8 gene; Immune; Immune response; Immunity; Immunocompetent; Immunocompromised Host; Immunologics; Individual; Infection; Inflammation Mediators; Inflammatory; Inhalation; Innate Immune Response; Investigation; Knowledge; Life Cycle Stages; Lung; Lung infections; Mediating; Methodology; Modeling; Monitor; Mus; Mycoses; Natural Resistance; Pathogenicity; Pathway interactions; Patients; Phagocytosis; Play; Proteins; Response Elements; Sampling; Signal Induction; Signal Pathway; Signal Transduction; Site; Soil; Sputum; Surface; System; T cell response; Tissues; Transgenic Organisms; United States; Work; airway epithelium; biosafety level 3 facility; chemokine; cytokine; experimental study; fungus; genome-wide; in vivo; innovation; insight; knockout gene; lentiviral-mediated; mortality; mouse model; new technology; novel; pathogen; pathogenic fungus; recruit; response; single-cell RNA sequencing; stem cells; transcriptome; transcriptomics; vaccine candidate

PROJECT SUMMARY Invasive fungal infections represent a major threat to immunocompromised patients and despite the availability of anti- fungal antibiotics, mortality rates remain as high as 50%. In the human host, the airway epithelium is the first point of contact upon inhalation of fungal conidia. As an immunologically active tissue, the airway epithelium may participate in active phagocytosis and coordinated immune cell recruitment. Soil fungi, including Coccidioides, do not need a human host to propagate, but upon disruption can enter the human body and lead to infection if not cleared. The fungal dimorph Coccidioides is native to arid regions in the southwestern U.S. and can establish infections in both immunocompetent and immunocompromised individuals. Unfortunately, little is known with regards to the initial response of human airway epithelium to Coccidioides. Currently, the Δcps1 strain of Coccidioides is being investigated as a potential vaccine candidate. Leveraging new technologies that permit the isolation of human airway stem cells, we demonstrated that primary differentiated human airway epithelial cells recapitulate the complexity of human airways. Our preliminary data showed infection at the apical surface of human airway epithelial cells by C. posadasii (Δcps1) elicits pro-inflammatory signals by the epithelium. Lastly, we demonstrated effective gene knockout in primary human airway epithelial cells using non-viral bulk nucleofection-based CRISPR strategy. Thus, our primary objective is to decipher the transcriptional and secretomal signatures critical to mount a successful response to Coccidioides in the human airway epithelium. To address our primary objective, we propose the following two specific aims: [1] determine the host response of human airway epithelium to both WT and the vaccine candidate C. posadasii (Δcps1), and [2] identify key signaling pathways to elicit an immune response to WT and the vaccine candidate C. posadasii (Δcps1) by human airway epithelium. This work will identify essential components of the human airway epithelium to mediate a protective response to these fungal infections and will provide the necessary foundation for future experiments to dissect the key pathways responsible for an effective host response to C. posadasii.

项目概要 尽管有抗真菌药物可用，但侵袭性真菌感染对免疫功能低下的患者构成了主要威胁。 真菌抗生素的死亡率仍高达50%。在人类宿主中，气道上皮是第一点 吸入真菌分生孢子后发生接触。气道上皮作为免疫活性组织，可能参与 主动吞噬作用和协调免疫细胞募集。土壤真菌，包括球孢子菌，不需要人类宿主 传播，但一旦受到破坏，如果不清除，可能会进入人体并导致感染。真菌二形体 球孢子菌原产于美国西南部的干旱地区，可以在免疫功能正常的人和有免疫力的人中引起感染。 免疫功能低下的个体。不幸的是，我们对人类呼吸道的最初反应知之甚少 上皮细胞为球孢子菌。目前，球孢子菌的 Δcps1 菌株正在作为一种潜在的疫苗进行研究 候选人。利用允许分离人类气道干细胞的新技术，我们证明了主要 分化的人类气道上皮细胞概括了人类气道的复杂性。我们的初步数据显示 C. posadasii (Δcps1) 对人气道上皮细胞顶端表面的感染可通过以下方式引发促炎信号： 上皮。最后，我们证明了使用非病毒在原代人气道上皮细胞中有效的基因敲除 基于批量核转染的 CRISPR 策略。因此，我们的主要目标是破译转录和分泌 这些特征对于在人类气道上皮细胞中对球孢子菌做出成功反应至关重要。为了解决我们的首要问题 出于目的，我们提出以下两个具体目标： [1] 确定人类气道上皮对两者的宿主反应 WT 和候选疫苗 C. posadasii (Δcps1) 以及 [2] 确定了引发免疫反应的关键信号通路 通过人气道上皮细胞将其转化为 WT 和候选疫苗 C. posadasii (Δcps1)。这项工作将确定必要的 人类气道上皮的成分介导对这些真菌感染的保护性反应，并将提供 为未来实验剖析负责宿主对艰难梭菌有效反应的关键途径奠定了必要的基础。 波萨达西。