Document text
Principal Investigator: Diane Bimczok
Organization: MONTANA STATE UNIVERSITY - BOZEMAN
Fiscal Year: 2021
Award: $462,348
Funding agency: National Institute of Biomedical Imaging and Bioengineering
Summary
The emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly infectious
human respiratory tract pathogen that has the capacity to infect many different organ systems, including the
gastrointestinal (GI) tract. The overall goal of this project is to elucidate the role of the GI mucosa in SARS-
CoV-2 infection and transmission. Specifically, we seek to identify mucosal immune mechanisms involved in
COVID-19 pathogenesis and compare epithelial responses to SARS-CoV-2 in the gut of humans and bats.
Investigating bat cells is important, because bats are considered the original hosts for SARS-CoV-2, but do not
develop overt disease upon infection. Our recently developed GOFlowChip platform, which integrates 3-
dimensional GI organoids and immune cells on a millifluidic chip with luminal and basolateral flow capacity, is
ideally suited to probe the mechanisms involved in SARS-CoV-2 infection of the gut mucosa. Here, we
propose to utilize the GOFlowChip to elucidate mechanisms of viral infection, replication and spread involved
in SARS-CoV-2 infection of the GI tract. Specifically, we seek to (1) Define how SARS-CoV-2 infection of the
GI epithelium contributes to viral spread. (2) Determine to what extent GI mucosal immune mechanisms
regulate SARS-CoV-2 infection and spreading in the GI tract. (3) Compare GI epithelial responses to SARS-
CoV-2 between bats and humans. For Specific Aim 1, we will optimize the GOFlowChip for use under BSL-3
laboratory conditions, followed by SARS-CoV-2 infection experiments that will elucidate susceptibility and viral
replication dynamics in different gut compartments. Specific Aim 2 will leverage our organoid-mononuclear
phagocyte (MNP) co-culture system to elucidate whether MNP-dependent transport mechanisms impact viral
invasion of the GI mucosa, and we will screen SARS-CoV-2 reactive patient sera for their ability to modulate
gut infection. For Specific Aim 3, we will establish organoid lines from bat GI tissues in order to compare
epithelial responses to SARS-CoV-2 in bats and humans using RNA sequencing in order to understand
differences in viral pathogenicity between humans and bats. The proposed work is directly integrated with our
existing project, because we will use out GOFlowChip to investigate how SARS-CoV-2 interacts with epithelial
cells and immune system components to cross the GI barrier. Our research is technologically innovative,
because we are using gut organoid-immune cell co-cultures in a fully contained tissue chip design to study a
BSL-3 level pathogen. Our project is conceptually innovative, because it is the first to compare gut organoids
from Jamaican fruit bats to human organoids for their ability to sustain SARS-CoV-2 infection and to mount
cellular antiviral responses. The proposed research is significant because it will yield crucial information on the
role of the GI mucosa in COVID-19 pathogenesis that may inform future strategies to identify viral carriers,
prevent viral transmission, and design novel antiviral treatments and vaccines.
Terms: <3-D><3-Dimensional><3D><Address><Alimentary Canal><Antigen-Presenting Cells><Antigens><Apical><Apoptosis><Apoptosis Pathway><Assay><Autoregulation><Bacteria><Bacterial Antigens><Bacterial Translocation><Bioassay><Biologic Assays><Biologic Models><Biological><Biological Assay><Biological Function><Biological Models><Biological Process><Biology><Biomedical Engineering><Biotech><Biotechnology><Blood monocyte><C pylori><C. pylori><Cadherin-1><Campylobacter pylori><Cell Body><Cell Communication><Cell Interaction><Cell-Extracellular Matrix><Cell-to-Cell Interaction><Cells><Co-culture><Cocultivation><Coculture><Coculture Techniques><Collaborations><Complex><Dendrites><Dendritic Cells><Development><Digestive Tract><Disease><Disorder><Drops><Drug Delivery><Drug Delivery Systems><E coli><E-Cadherin><E. coli><ECM><Engineering><Epithelial><Epithelial Calcium-Dependent Adhesion Protein><Epithelial Cells><Epithelial-Cadherin><Escherichia coli><Extracellular Matrix><Fc Receptor><FcRn><FcRn neonatal transfer protein><GI Tract><GI microbiota><Gastrointestinal Tract><Gastrointestinal microbiota><Gastrointestinal tract structure><Gut Epithelium><H pylori><H pylory><H. pylori><H. pylory><Health><Helicobacter pylori><Homeostasis><Human><Human Experimentation><Human Microbiome><Image><Immune><Immune Cell Activation><Immune Tolerance><Immune system><Immunes><Immunity><Immunologic Tolerance><Immunology><Intestinal><Intestines><Investigation><Investigators><Liquid substance><Marrow monocyte><Microbe><Microbiology><Microbiomics><Microfluidics><Model System><Modeling><Modern Man><Mononuclear><Movement><Mucosa><Mucosal Immunity><Mucosal Tissue><Mucous Membrane><O element><O2 element><Occluding Junctions><Organoids><Oxygen><Phagocytes><Phagocytic Cell><Physiological Homeostasis><Population><Position><Positioning Attribute><Programmed Cell Death><RNA Seq><RNA sequencing><RNAseq><Reproducibility><Research><Research Personnel><Researchers><Sampling><Sentinel><Side><Sorting - Cell Movement><Stomach><Surface><System><Testing><Therapeutic><Thick><Thickness><Tight Junctions><Time><Uvomorulin><Veiled Cells><Walking><Work><Zonula Occludens><accessory cell><adaptive immunity><alimentary tract><allergic/immunologic body system><allergic/immunologic organ system><amebocyte><antibody receptor><base><beneficial flora><beneficial microbes><beneficial microflora><beneficial microorganism><bio-engineered><bio-engineers><bioengineering><body movement><bowel><cell type><clinical applicability><clinical application><deliver vaccines><design><designing><develop a vaccine><development of a vaccine><developmental><digestive canal><enteric microbial community><enteric microbiota><field based data><field learning><field study><field test><fluid><fluid flow><gastric><gastric organoids><gastroid><gastrointestinal><gastrointestinal epithelium><gastrointestinal homeostasis><gastrointestinal microbial flora><gut commensal><gut community><gut flora><gut microbe community><gut microbes><gut microbial community><gut microbial composition><gut microbial consortia><gut microbial species><gut microbiota><gut microbiotic><gut microflora><host microbe association><host microbe relationship><host-microbe interactions><host-microbial interactions><host-microorganism interactions><human tissue><human-associated microbiome><imaging><immune activation><immune system tolerance><immune unresponsiveness><immunogen><immunological paralysis><in vitro Model><inhibitor><inhibitor/antagonist><innovate><innovation><innovative><intestinal flora><intestinal homeostasis><intestinal microbes><intestinal microbiota><intestinal microflora><intestinal tract microflora><liquid><matrigel><mechanosensing><mechanotransduction><member><microbe pathogen><microbial><microbial antigen><microbial colonization><microbial consortia><microbial flora><microbial pathogen><microbiome><microbiome research><microbiome science><microbiome studies><microbiota><microflora><microorganism><microorganism antigen><migration><monocyte><mucosal vaccine><multispecies consortia><neonatal Fc receptor><neutralizing antibody><new vaccines><next generation vaccines><novel><novel vaccines><organ on a chip><organ on chip><pathogenic microbe><prototype><recruit><residence><residential building><residential site><response><shear stress><sorting><stomach organoid><temporal measurement><temporal resolution><therapeutic agent development><therapeutic development><three dimensional><time measurement><tissue culture><tool><transcriptome sequencing><transcytosis><uptake><vaccine delivery><vaccine development><vaccine formulation><µfluidic>