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.

项目摘要 Covid-19的发病率和死亡率是夸张的炎症反应导致严重组织的结果和器官损坏。但是，为什么某些人从最初稳定中发展的生物学机制最终尚未完全阐明危险条件。这项工作提出了免疫反应最早感染阶段的感染部位在随后的病理学上起着确定性的作用；即在呼吸道粘膜中的延迟或抑制1型干扰素反应在呼吸道粘膜中感染允许快速病毒增殖和较小的症状，但最终导致了高病毒载荷，并且后来在疾病中看到的夸张炎症反应。此外，拟议的项目将测试是否延迟干扰素反应是先前记录的与年龄相关功能障碍的结果。它也试图确定直接SARS-COV-2是否介导的宿主剪接破坏也可以抑制干扰素在感染的早期阶段发出信号。为此，该研究将利用一组独特的纵向配对来自最初对SARS-COV-2感染负阴性的个体的鼻拭子和唾液样本，但成为作为IRB批准的一部分，被前瞻性采样时被感染（每天两次）（＃20- 1026）申请人共同设计和共同领导者的共同传播研究（自2020年8月以来）在加利福尼亚理工学院。这些样品中存在的转录组将进行分析以测量基因和同工型表达，白细胞募集和激活，包括通过干扰素可以通过难以捉摸的感染早期阶段和疾病的全部病程来推断信号。年轻时6岁的患者的纵向差分表达和差剪接路径将评估年龄以确定与年龄相关的免疫反应差异（特别是干扰素）信号传导）导致病毒负荷和随后的症状严重程度的更快增加。另外，来自RNA 将富含新生前MRNA的测序文库，将测量剪接缺陷，例如内含子保留，确定病毒介导的剪接破坏是否也导致抑制或延迟的早期干扰素允许快速病毒增殖的反应。 Covid-19是公共卫生威胁，并且与任务一致在NIAID中，这项研究的结果可以提供有关SARS-COV-2的病理生理学的机械见解感染，有可能识别预防或治疗严重疾病的新颖或更有效的靶标。此外，拟议项目为申请人提供了极好的培训机会，以获取知识和免疫学，病毒学/冠状病毒生物学，人RNA剪接机制和生成分析的技能 Rustem Ismagilov博士，Akiko Iwasaki博士和米奇·古特曼（Mitch Guttman）博士是上述领域的专家。