权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Intestinal organoid modeling of SARS-CoV-2-stimulated innate and adaptive immunity

SARS-CoV-2 刺激的先天性和适应性免疫的肠道类器官模型

基本信息

批准号：
10319858
负责人：
CALVIN J KUO
金额：
$ 39.36万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-15 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10319858
关键词：
2019-nCoV 3-Dimensional ACE2 Acute Air Antibodies Antiviral Response Apical B-Cell Antigen Receptor B-Lymphocytes Bacterial Infections Biological Models Biopsy COVID-19 mortality COVID-19 pandemic COVID-19 patient Cell Communication Cells Clinical Coculture Techniques Collaborations Colon Common Cold Communication Complex Convalescence Coronavirus Coronavirus Infections Data Diarrhea Epithelial Epithelial Cells Event Fostering Gluten Human Immune Immune checkpoint inhibitor Immune response Immune system Immunity Immunization Immunize Immunological Models In Vitro Incubators Infection Inflammation Innate Immune Response Intervention Intestines Investigation Liquid substance Lung Lung diseases Lung infections Malignant Neoplasms Medicine Methods Modeling Myelogenous Myeloid Cells Natural Immunity Nature Nodal Organ Organoids Pathogenesis Pathology Patients Peripheral Pharmacology Plasma Public Health Receptor Cell Reporting Respiratory Failure SARS-CoV-2 infection SARS-CoV-2 pathogenesis Serology Small Intestines Specificity Surface Surveys Suspension Culture Suspensions Symptoms System T-Cell Receptor T-Lymphocyte Technology Testing Therapeutic Time Tissue Model Tissue Preservation Tissues Tonsil Transcript Tumor-infiltrating immune cells Type 2 Angiotensin II Receptor Variant Virus Diseases Virus Replication Vomiting adaptive immune response adaptive immunity cell type co-infection cross reactivity cytokine release syndrome enteric infection global health health economics immunoregulation in vitro Model intestinal epithelium lymph nodes mesenteric lymph node network models novel novel coronavirus novel therapeutics reconstitution response single cell analysis single-cell RNA sequencing systemic inflammatory response therapeutic development therapeutic evaluation three dimensional cell culture tumor

项目摘要

ABSTRACT The COVID-19 pandemic, engendered by the novel coronavirus SARS-CoV-2, is a grave threat to public health, with lung infection and respiratory failure. However, the intestine is also targeted by SARS-CoV-2, as many patients present with GI symptoms and the SARS-CoV-2 receptor angiotensin-converting enzyme 2 (ACE2) is abundantly expressed by intestinal epithelium. COVID-19 mortality is strongly associated with systemic inflammation as in “cytokine storm”, fostering attempts at therapeutic immunomodulation, and raising a critical need for in vitro human systems modeling SARS-CoV-2-induced immune cell interactions with intestinal epithelium. Conventional 3D organoid cultures (“enteroids”) allow intestinal epithelial SARS-CoV-2 infection but unfortunately omit immune cells. Here we generate a holistic intestinal in vitro model of SARS-CoV-2 infection using air-liquid interface (ALI) organoids containing both epithelium and infiltrating immune cells en bloc without artificial reconstitution. The complex intestinal immune system of ALI intestinal organoids contains various innate and adaptive immune cells, highly diverse T cell receptor (TCR) and B cell receptor (BCR) repertoire, and plasma B cell-derived antibody transcripts. Importantly, immune components in these ALI organoids respond efficiently to epithelial damage and ALI intestinal organoids are highly susceptible to bacterial and viral infections. Here, we utilize this unique ALI organoid technology with integrated immune components to explore sequelae of SARS-CoV-2 infection. Aim 1 establishes and optimizes BSL3 SARS-CoV-2 infection of ALI intestinal organoids, exploiting a novel eversion method to relocate the apical aspect of ACE2-expressing cells to the external surface, allowing survey of SARS-CoV-2 infection of different regions of small intestine and colon. The time course of tissue-resident SARS-CoV-2-induced cross-talk between intestinal epithelium and immune cells is unknown, as hindered by lack of human experimental systems. Thus, Aim 2 performs a scRNA- seq and CyTOF study of SARS-CoV-2-induced immune responses within ALI organoids to (1) create a network model of the temporal propagation of immunity and bidirectional communication between epithelium and immune cells and (2) perform therapeutic testing against nodal vulnerabilities, correlating against clinical status (naïve, convalescent, immunized, cross-reactive coronavirus). Current epithelial organoid systems also do not allow study of adaptive immunity. Aim 3 thus recapitulates SARS-CoV-2 adaptive immune responses by co-culturing ALI intestinal organoids with newly developed human lymph node organoids. Within such SARS-CoV-2-infected co-cultures we correlate adaptive immune responses with clinical status including prior cross-reactive coronavirus infection and SARS-CoV-2 naïve, convalescent and ultimately immunized states. Overall, we leverage collaboration from Mark Davis (human LN culture) and Catherine Blish and Scott Boyd (SARS-CoV-2) to create human in vitro systems modeling SARS-CoV-2-induced innate and adaptive immunity, with relevance for pathogenesis investigations and therapeutics testing.

摘要