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Experimental and natural SARS-CoV-2 infection of the human pancreas

人类胰腺的实验性和自然 SARS-CoV-2 感染

基本信息

批准号：
10319841
负责人：
DIRK HOMANN
金额：
$ 45.33万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
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：
2021-09-17 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10319841
关键词：
2019-nCoV ACE2 Address Animal Experiments Area Autopsy Beta Cell Biological Biomedical Research Blood Glucose COVID-19 COVID-19 mortality COVID-19 patient Cell physiology Cells Clinical Cohort Studies Cytometry Data Detection Diabetes Mellitus Endocrine Epidemiology Experimental Designs Flow Cytometry Foundations Future Generations Glucose Glucose tolerance test Goals Histologic Histology Human Immune Immunodeficient Mouse Immunologics Impairment In Situ In Vitro Infection Insulin Insulin-Dependent Diabetes Mellitus Intervention Investigation Islets of Langerhans Islets of Langerhans Transplantation Journals Letters Logistics Mediating Medicine Messenger RNA Metabolic Modeling Mus Nature New England Non-Insulin-Dependent Diabetes Mellitus Pancreas Pathologic Pathway interactions Pattern Physiological Plant Roots Plasma Population Predisposition Proteins Proteome Protocols documentation Publications Publishing Reporting Research Research Personnel Resolution Risk Assessment Role SARS-CoV-2 infection Sampling Series Severities Slice Stains Therapeutic Intervention Time Tissues Tropism Viral Viral Load result Viral Proteins Virus Diseases Work base blood glucose regulation case control cell type comorbidity demographics diabetogenic epidemiologic data experimental study high dimensionality human model humanized mouse in vitro Model in vivo influenza infection insight insulin secretion islet mRNA Expression mouse model non-diabetic patient registry prophylactic receptor response transcriptome virology

项目摘要

ABSTRACT The possibility that infection with SARS-CoV-2 may trigger diabetes emerged as a concern in mid 2020. We have addressed this issue by developing a research roadmap that identified complementary areas of investigation according to several broad topics in the realm of relevant basic biomedical research. Based on our recent publication about the expression of SARS-CoV-2 entry factors in the non-diabetic human pancreas, we have now developed a Research Strategy that will assess the principal capacity of pancreatic SARS-CoV-2 infection and its primary consequences in a series of in vitro, in vivo and ex vivo studies distributed across three Specific Aims: In Aim 1 we will define expression patterns of ACE2 and other viral entry factors in dispersed human pancreatic cell populations; will subject islets to in vitro SARS-CoV-2 infection and quantify infection patterns as well as alterations of single-cell proteomes and transcriptomes by flow cytometry, high-dimensional mass cytometry, and scRNAseq; will determine functional islet responses under conditions of viral infection by dynamic glucose-stimulated insulin secretion; will conduct mechanistic studies with targeted ACE2 blockade and by generation of ACE2-deficient beta-like cells; and will explore SARS-CoV-2 infection in the context of living pancreas slices that provide a more “physiological” experimental platform for in vitro studies. In Aim 2, we will use the robust “minimal mass” model of human islet transplantation into immunodeficient mice to delineate metabolic perturbations (blood glucose values, plasma insulin, intraperitoneal glucose tolerance test) accrued in the wake of an in vivo SARS-CoV-2 challenge. To address the considerable experimental and logistical challenges in these studies, we have developed a tiered approach that progressively refines experimental designs (including use of a recently generated, sequenced and characterized mouse-adapted [ma] SARS-CoV-2 strain) with the goal to imbue our models with escalating biological relevance. In Aim 3, we will draw on our access to COVID-19 and control autopsy cases to reveal the ex vivo viral burden of natural SARS-CoV-2 infection in the pancreas. These studies, which build on recently published pilot data as well as extensive preparatory studies to adjust our staining protocols to the demands of at times suboptimal tissue quality, will define SARS-CoV-2 protein and mRNA abundance, cellular association and distribution patterns across the COVID-19 pancreas. We will further leverage an established immunohistochemical multiplexing strategy to define the identity, quantity and distribution of major immune cell subsets throughout COVID-19 and control pancreata. Thus, the proposed work will generate an inclusive perspective on the cardinal viral and immunological components of the potentially altered pancreatic histology in COVID-19. Collectively, the proposed work addresses, and is expected to resolve at least in part, key aspects of the hypothesis that SARS-CoV-2 infection may precipitate diabetes onset. Together with emerging epidemiological data, it may therefore provide an important foundation for future risk assessment and the prioritization of prophylactic and/or therapeutic intervention strategies.

摘要