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Principal Investigator: Deok-Ho Kim
Organization: CORNELL UNIVERSITY
Fiscal Year: 2024
Award: $237,913
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY
Blood vessels play a central role in maintaining host immunity by transporting immune cells to sites of infection.
During the process, blood vessels experience endothelial junction remodeling to control vascular permeability
and immune cell extravasation. Under infection, blood vessels become permeable and allow immune cells to
extravasate and kill pathogens in the interstitium. Once the infection is resolved, permeable vessels become less
permeable and limit the number of interstitial immune cells. However, sometimes in inflammation, the remodeling
is perturbed, resulting in prolonged, hyper-permeable blood vessels. This vascular dysfunction contributes to
immune diseases, such as chronic inflammation, lupus, and autoimmune disease. It is known that endothelial
cell alignment is crucial to maintain intact cell-cell adhesion and promote junction maturation. Despite the
significance of the cell alignment in functional endothelium, currently available high-throughput methods, such
as real-time cell analysis (RTCA) and trans-epithelial/trans-endothelial electrical resistance (TEER) systems with
randomly seeded cells have not successfully measured cell impedance or electrical resistance through the in
vivo-like controlled endothelial cell morphology, alignment, and matured cell-cell junctions. Furthermore, the
current technologies lack pericyte co-culture with endothelial cells. In this proposal, we will develop a high-
throughput, high-content functional screening assay capable of faster drug screening and mechanistic studies
on blood vessel barrier function and immune cell extravasation. To achieve our goals, we will establish a
nanopatterned IEA-based functional assay for high-throughput phenotype screening of pericyte-covered
endothelium. To establish the nanopatterned IEA-based assay, we will determine conditions for junctional
maturation of human dermal and lung microvascular endothelial cells with or without pericytes, focusing on (i)
degree of cell alignment; (ii) expression of adherens junctions, polarization, and basement membrane markers;
(iii) vascular barrier function (Aim 1.1). We will then assess vascular gene expression profiles related to vessel
stabilization and immune cell adhesion. We will next evaluate immune cell extravasation through the endothelium
in the non-inflammatory condition to determine immune cell behaviors in steady-state blood vessels (Aim 1.2).
Next, we will validate the utility of the system for inflammation-induced blood vessel dysfunction. To achieve this
aim, we will examine the endothelial barrier function and immune cell extravasation in five different categories of
inflammatory cytokines and various levels of substrate stiffness considering skin and lung microenvironments
(Aim 2.1). Lastly, we will identify potential targets and drugs to reverse vessel dysfunction by focusing on
abrogation of the cytokine effect and the stiffness effect, separately or in combination (Aim 2.2). In summary, our
system will constitute a significant improvement over existing technologies as it represents a novel high-
throughput screening tool for functionally matured blood endothelium and their interactions with immune cells.
Terms: <Address><Adherens Junction><Adhering Junction><Adhesive Junction><Adventitial Cell><Anchoring Junction><Animal Genetics><Animal Model><Animal Models and Related Studies><Animals><Assay><Autoimmune Diseases><Basement membrane><Bioassay><Biological Assay><Biology><Blood><Blood - brain barrier anatomy><Blood Reticuloendothelial System><Blood Vessels><Blood-Brain Barrier><COVID-19><CV-19><Categories><Cell Adhesion><Cell Body><Cell Communication><Cell Communication and Signaling><Cell Interaction><Cell Junctions><Cell Locomotion><Cell Migration><Cell Movement><Cell Signaling><Cell-Cell Adhesion><Cell-to-Cell Interaction><Cells><Cellular Adhesion><Cellular Migration><Cellular Morphology><Cellular Motility><Chronic><Clotting><Co-culture><Coagulation><Coagulation Process><Cocultivation><Coculture><Coculture Techniques><Communicable Diseases><Coronavirus Infectious Disease 2019><Deposit><Deposition><Dermal><Disease><Disorder><Drug Screening><Drugs><Dysfunction><Electric Resistance><Electrical Impedance><Electrical Resistance><Electrodes><Endothelial Cells><Endothelium><Epithelium><Experimental Models><Expression Signature><Extravasation><Functional disorder><Gene Expression><Gene Expression Profile><Goals><Hemato-Encephalic Barrier><High Throughput Assay><Human><Immune><Immune Diseases><Immune Disorders><Immune Dysfunction><Immune System Diseases><Immune System Disorder><Immune System Dysfunction><Immune System and Related Disorders><Immunes><Immunity><Immunodeficiency and Immunosuppression Disorders><Immunologic Diseases><Immunological Diseases><Immunological Dysfunction><Immunological System Dysfunction><Immunology><Impairment><Impedance><In Vitro><Infection><Infectious Disease Pathway><Infectious Diseases><Infectious Disorder><Inflammation><Inflammatory><Intercellular Junctions><Intracellular Communication and Signaling><Investigation><Leakage><Lesion><Lung><Lung Respiratory System><Lupus><Lymphatic><Measurement><Measures><Mediating><Medication><Methods><Modeling><Modern Man><Nature><Pattern><Pericapillary Cell><Pericytes><Perivascular Cell><Permeability><Pharmaceutical Preparations><Phenotype><Physiologic><Physiological><Physiopathology><Play><Process><Rapid screening><Reading><Role><Rouget Cells><Screening procedure><Sepsis><Signal Transduction><Signal Transduction Systems><Signaling><Site><Skin><Spillage><System><Systems Analyses><Systems Analysis><Technology><Therapeutic><Time><Vascular Diseases><Vascular Disorder><Vascular Permeabilities><autoimmune condition><autoimmune disorder><autoimmunity disease><biological signal transduction><blood infection><blood vessel disorder><bloodbrain barrier><bloodstream infection><cell behavior><cell morphology><cell motility><cellular behavior><clinical relevance><clinically relevant><coronavirus disease 2019><coronavirus disease-19><coronavirus infectious disease-19><cytokine><drug candidate><drug/agent><electric impedance><endothelial dysfunction><experience><gene expression pattern><gene expression signature><glomerular filtration><high throughput screening><iPS><iPSC><iPSCs><improved><in vitro Model><in vivo><induced pluripotent cell><induced pluripotent stem cell><inducible pluripotent stem cell><interstitial><knock-down><knockdown><lung microvascular endothelial cells><lung vascular endothelial cells><migration><model of animal><nano pattern><nanopattern><novel><pathogen><pathophysiology><personalization of treatment><personalized medicine><personalized therapy><personalized treatment><polarized cell><prevent><preventing><pulmonary><pulmonary microvascular endothelial cells><pulmonary vascular endothelial cells><screening><screening tools><screenings><social role><trafficking><transcriptional profile><transcriptional signature><vascular><vascular dysfunction><vasculopathy>