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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要 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，为什么有些个体从最初的稳定状态发展到了一个新的阶段，其生物学机制最终危急情况尚未完全阐明。这项工作提出，免疫反应感染最早阶段的感染部位对随后的病理学起着决定性作用；即在治疗的最初几天内，呼吸道粘膜中的 1 型干扰素反应延迟或受到抑制感染允许病毒快速增殖，症状轻微，但最终导致高病毒载量和疾病后期出现的过度炎症反应。此外，拟议的项目将测试是否延迟的干扰素反应是先前记录的与年龄相关的功能障碍的结果。它还力求确定直接 SARS-CoV-2 介导的宿主剪接破坏是否也能抑制干扰素感染早期阶段的信号传导。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来自最初对 SARS-CoV-2 感染呈阴性但后来变成阴性的个体作为 IRB 批准的一部分 (#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。这些样本中存在的转录组将被分析以测量白细胞募集和激活的基因和亚型表达，包括通过干扰素通过难以捉摸的感染早期阶段和整个疾病过程可以推断出信号传导。 6 岁患者和晚期患者的纵向差异表达和差异剪接路径将评估年龄，以确定免疫反应（特别是干扰素信号）导致病毒载量更快增加，以及随后的症状严重程度。此外，从 RNA 富含新生前体 mRNA 的测序文库，将测量内含子保留等剪接缺陷，确定病毒介导的剪接破坏是否也会导致早期干扰素的抑制或延迟反应允许病毒快速增殖。 COVID-19 是一种公共卫生威胁，符合我们的使命 NIAID 的研究结果可以为 SARS-CoV-2 的病理生理学提供机制见解感染，以潜在地确定预防或治疗严重疾病的新的或更有效的目标。此外，拟议的项目为申请人提供了一个极好的培训机会，以获取知识和免疫学、病毒学/冠状病毒生物学、人类 RNA 剪接机制和生成分析技能在 Rustem Ismagilov 博士、Akiko Iwasaki 博士的指导下，对 RNA 测序数据进行解释和解释 Mitch Guttman 博士是上述领域的专家。