Longitudinal mucosal immune response to SARS-CoV-2 starting prior to infection

在感染前就开始对 SARS-CoV-2 进行纵向粘膜免疫反应

• 批准号: 10538180
• 负责人: Alexander Viloria Winnett
• 金额: $ 5.18万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10538180
• 关键词: 2019-nCoV; Accounting; Acute; Adult; Age; Antiviral Response; Autopsy; Biological; Biology; COVID-19; California; Cell Culture Techniques; Characteristics; Child; Clinical; Collaborations; Coronavirus; Data; Data Set; Defect; Development; Disease; Disease Progression; Elderly; Enrollment; Frequencies; Functional disorder; Generations; Genes; Genetic Transcription; Household; Human; Immune; Immune System Diseases; Immune response; Immunobiology; Immunology; Individual; Infection; Inflammatory; Inflammatory Response; Influenza; Institutes; Institutional Review Boards; Interferon Suppression; Interferon Type II; Interferons; Introns; Kinetics; Knowledge; Laboratories; Lead; Leukocytes; Libraries; Measurement; Measures; Mediating; Mentorship; Messenger RNA; Minor; Mission; Morbidity - disease rate; Mucosal Immune Responses; Mucous Membrane; National Institute of Allergy and Infectious Disease; Nonsense-Mediated Decay; Nonstructural Protein; Nucleic Acids; Obesity; Ontology; Organ; Pathology; Pathway interactions; Patients; Phase; Play; Prevention; Prophylactic treatment; Protein Isoforms; Public Health; RNA Splicing; Research Project Grants; Respiratory Mucosa; Risk Factors; Role; SARS-CoV-2 infection; SARS-CoV-2 negative; Saliva; Sampling; Severities; Severity of illness; Signal Transduction; Site; Specimen; Symptoms; Technology; Testing; Tissue Sample; Tissues; Transcript; Translations; Treatment Effectiveness; Viral; Viral Load result; Viral load measurement; Virus; Work; acute infection; age related; computerized tools; design; differential expression; high risk; immunoregulation; insight; interest; mRNA Precursor; mRNA sequencing; mortality; nasal swab; novel; prospective; protein transport; recruit; repository; respiratory; response; saliva sample; severe COVID-19; skills; temporal measurement; therapeutic target; training opportunity; transcriptome; transcriptome sequencing; transcriptomics; transmission process; virology

PROJECT SUMMARY Morbidity and mortality in COVID-19 is the result of an exaggerated inflammatory response causing severe tissue and organ damage. However, the biological mechanism for why some individuals progress from initially stable to ultimately critical condition has not been fully elucidated. This work proposes that the immune response at the site of infection during the earliest stage of infection plays a deterministic role on subsequent pathology; namely that a delayed or suppressed type 1 interferon response in the respiratory mucosa within the first few days of infection permits rapid viral proliferation with minor symptoms, but eventually leads to the high viral loads and exaggerated inflammatory response seen later in disease. Further, the proposed project will test whether a delayed interferon response is the result of previously documented age-related dysfunction. It also seeks to determine whether direct SARS-CoV-2 mediated disruption of host splicing can also suppress interferon signaling in the early stage of infection. To do this, the study will leverage a unique set of longitudinal paired nasal swab and saliva samples from individuals who are initially negative for SARS-CoV-2 infection, but become infected while being prospectively sampled with high frequency (twice per day) as part of an IRB-approved (#20- 1026) COVID-19 household transmission study that the applicant co-designed and co-leads (since August 2020) at the California Institute of Technology. The transcriptome present in these samples will be analyzed to measure gene and isoform expression from which leukocyte recruitment and activation, including through interferon signaling can be inferred, through the elusive early stage of infection and the full course of the illness. Longitudinal differential expression and differential splicing paths in patients as young as age 6, and of advanced age will be assessed to identify whether age-related differences in immune response (in particular, interferon signaling) lead to more rapid increases in viral load, and subsequent symptom severity. In addition, from an RNA sequencing library enriched for nascent pre-mRNAs, splicing defects such as intron retention will be measured, to identify whether virus-mediated disruption of splicing also leads to a suppressed or delayed early interferon response permissive of rapid viral proliferation. COVID-19 is a public health threat, and in line with the mission of the NIAID, the results of this study can provide mechanistic insights into the pathophysiology of SARS-CoV-2 infection, to potentially identify novel or more efficient targets for the prevention or treatment of severe disease. In addition, the proposed project offers an excellent training opportunity for the applicant to gain knowledge and skills in immunology, virology/coronavirus biology, mechanisms of human RNA splicing, and generation analysis and interpretation of RNA sequencing data, with mentorship from Dr. Rustem Ismagilov, Dr. Akiko Iwasaki, and Dr. Mitch Guttman, who are experts in the aforementioned fields.

项目摘要 COVID-19的发病率和死亡率是过度炎症反应导致严重组织损伤的结果。 和器官损伤然而，为什么有些人从最初的稳定进展的生物机制， 到最后的临界状态还没有完全阐明。这项工作提出，免疫反应在 在感染的最早阶段的感染部位对随后的病理学起决定性作用;即 在最初几天内，呼吸道粘膜中延迟或抑制的1型干扰素应答 感染允许病毒快速增殖，症状轻微，但最终导致高病毒载量， 在疾病后期观察到的过度炎症反应。此外，拟议的项目将测试 延迟的干扰素应答是先前记录的年龄相关功能障碍的结果。它还寻求 确定直接SARS-CoV-2介导的宿主剪接破坏是否也能抑制干扰素 在感染的早期阶段。为此，该研究将利用一组独特的纵向配对 鼻拭子和唾液样本，这些样本来自最初对SARS-CoV-2感染呈阴性的个体，但后来成为 感染，同时作为IRB批准的（#20- 1026）申请人共同设计和共同领导的COVID-19家庭传播研究（自2020年8月起） 在加州理工学院。将分析这些样本中存在的转录组，以测量 白细胞募集和激活的基因和同种型表达，包括通过干扰素 信号可以通过感染的早期阶段和疾病的整个过程来推断。 6岁及晚期乳腺癌患者的纵向差异表达和差异剪接途径 将评估年龄以确定是否存在免疫应答（特别是干扰素）的年龄相关差异 信号传导）导致病毒载量的更快速增加和随后的症状严重性。此外，从RNA 测序富集新生前mRNA的文库，将测量剪接缺陷如内含子保留， 确定病毒介导的剪接破坏是否也导致抑制或延迟早期干扰素 允许病毒快速增殖的反应。COVID-19是一种公共卫生威胁，符合使命 的NIAID，这项研究的结果可以提供机制的见解SARS-CoV-2的病理生理学 感染，以潜在地鉴定用于预防或治疗严重疾病的新的或更有效的靶标。 此外，建议的项目为申请人提供了一个极好的培训机会， 免疫学、病毒学/冠状病毒生物学、人类RNA剪接机制和世代分析技能 和RNA测序数据的解释，从博士Rustem Ismagilov，博士Akiko Iwasaki，和 博士Mitch Guttman是上述领域的专家。