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Principal Investigator: ANNE E CRESS
Organization: UNIVERSITY OF FLORIDA
Fiscal Year: 2024
Award: $378,584
Funding agency: National Heart Lung and Blood Institute
ABSTRACT:
The elevated ARDS mortality observed in the COVID-19 pandemic has highlighted the contribution of excessive
mechanical stress produced by mechanical ventilation in promoting lethal increases in lung vascular permea-
bility. Integrin β4 (ITGB4 gene) and kindlin-2 (FERMT2 gene) are essential adhesion molecules in endothelial
cell (EC) focal adhesions (FAs), structures critical for mechano-sensing, for bidirectional signaling between
the EC cytoskeleton and the cell-matrix interface, and for EC barrier regulation. The mechanistic basis for dy-
namic FA coordination during inflammatory EC barrier dysfunction and subsequent barrier restoration is a fun-
damental question that remains unresolved. We speculate that coordinate control of FA structures requires
the dynamic interactions of integrin β4 (ITGB4) and kindlin-2 with key PPG cytoskeletal effectors (nmMLCK,
cortactin, Dock1, lamellipodin, paxillin) to efficiently assemble functional FAs during EC barrier responses (pe-
ripheral cytoskeletal remodeling, lamellipodial formation, gap closure). ITGB4 is a unique mechano-sensing,
laminin-binding integrin, and kindlin-2 is a multi-domain mechano-sensing adapter FA protein that recruits struc-
tural and signaling molecules to FAs in concert with cytoskeletal rearrangement. We speculate that these EC
responses are highly influenced by ITGB4 and kindlin-2 post-translational modifications (PTMs) and coding pol-
ymorphisms (SNPs). As reactive oxygen species (ROS) is an important stimulus for FA dynamics and loss of
EC barrier integrity, with Core B, SA #1 will characterize the role of three ROS-sensing transcription factors
(NRF2, HIF1α/HIF2α), ITGB4/FERMT2 SNPs and DNA methylation in genetic/epigenetic regulation of
ITGB4/FERMT2 expression and the influence on generation of the unique ITGB4 alternatively-spliced, barrier-
regulatory variant, Integrin β4E (ITGB4E), that we identified as involved in mechano-sensing and EC barrier
regulation. With Core D, SA #2 will conduct in depth structure/function studies including 3D live cell imaging of
mutant ITGB4 and kindlin-2 fusion proteins (SNPs, PTMs) to characterize ITGB4/kindlin-2 function in spatially-
specific EC cytoskeletal rearrangements driving EC barrier-disruption and barrier-restoration. SA #3 will examine
the functionality of ITGB4/kindlin-2 interactions within lamellipodia with known and novel FA-binding cytoskeletal
partners, including cortactin, Dock1, lamellipodin and highly novel interactions with nmMLCK (Project #1).
Finally, utilizing elegant rat and porcine models of LPS/VILI (Core C), SA #4 will assess the therapeutic utility of
the SMURF inhibitor A01, or SRI-38832 to augment kindlin-2 expression as cargo in TySIPonate-conjugated
liposomes (TySIPosomes, Project #4). Project #3 studies will determine the structure/function and molecular
basis for the dynamic FA control by ITGB4 and kindlin-2 and yield important insights into functional relevance of
this unique FA signaling axis in restoration of EC barrier function. Our highly translational approaches will also
provide actionable EC barrier-regulatory strategies that restore the integrity of the injured pulmonary circulation
while yielding insights into ITGB4/FERMT2 variant participation in ARDS health disparities.
Terms: <3-D><3-Dimensional><3D><500-kDa HD1 protein><ARDS><Acceleration><Actins><Active Oxygen><Acute Respiratory Distress><Acute Respiratory Distress Syndrome><Address><Adhesion Molecule><Adhesion Plaques><Adhesions><Adult ARDS><Adult RDS><Adult Respiratory Distress Syndrome><Alternate Splicing><Alternative RNA Splicing><Alternative Splicing><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-inflammatory><Automobile Driving><Basal Transcription Factor><Basal transcription factor genes><Binding><Blood Vessels><CD104 Antigens><COVID crisis><COVID epidemic><COVID pandemic><COVID-19 crisis><COVID-19 epidemic><COVID-19 era><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 pandemic><COVID-19 period><COVID-19 public health crisis><COVID-19 years><CTTN><CTTN Gene><Cell Adhesion Molecule Gene><Cell Adhesion Molecules><Cell Body><Cell Communication and Signaling><Cell Junctions><Cell Signaling><Cell membrane><Cell-Matrix Adherens Junctions><Cells><Cellular Matrix><Cellular Mechanotransduction><Chimera Protein><Chimeric Proteins><Code><Coding System><Common Rat Strains><Complex><Cortactin Gene><Cytoplasmic Domain><Cytoplasmic Membrane><Cytoplasmic Tail><Cytoskeletal Gene><Cytoskeletal Proteins><Cytoskeletal System><Cytoskeleton><DNA Methylation><Da Nang Lung><Dysfunction><E3 Ligase><E3 Ubiquitin Ligase><EMS1><EMS1 gene><Endothelial Cells><Endothelium><Erinaceidae><FADK><FAK><FAK1><Family suidae><Focal Adhesions><Focal Contacts><Functional disorder><Fusion Protein><General Transcription Factor Gene><General Transcription Factors><Generations><Genes><Genetic><Genetic Polymorphism><Glycoprotein GP-2><HIF 1 alpha><HIF-1alpha><HIF1-Alpha><HIF1A><HIF1A gene><HIF1α><Hedgehogs><Inflammation><Inflammatory><Integrin Binding><Integrin beta(4)><Integrin beta4><Integrin β4><Integrins><Integrins Extracellular Matrix><Intercellular Junctions><Intracellular Communication and Signaling><Laminin><Liposomal><Liposomes><Lung><Lung Respiratory System><Lung damage><MOF syndrome><MOP1><Mechanical Signal Transduction><Mechanical Stress><Mechanical ventilation><Mechanosensory Transduction><Mediating><Modeling><Molecular><Molecular Interaction><Multiple Organ Dysfunction Syndrome><Multiple Organ Failure><Oxygen Radicals><PTK2><PTK2 gene><Peripheral><Phosphorylation><Physiopathology><Pigs><Plasma Membrane><Post-Translational Modification Protein/Amino Acid Biochemistry><Post-Translational Modifications><Post-Translational Protein Modification><Post-Translational Protein Processing><Posttranslational Modifications><Posttranslational Protein Processing><Pre-Clinical Model><Preclinical Models><Pro-Oxidants><Protein Modification><Protein Phosphorylation><Proteins><Pulmonary Circulation><Rat><Rats Mammals><Rattus><Reactive Oxygen Species><Regulation><Role><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 pandemic><SARS-coronavirus-2 epidemic><SARS-coronavirus-2 pandemic><Severe Acute Respiratory Syndrome CoV 2 epidemic><Severe Acute Respiratory Syndrome CoV 2 pandemic><Severe acute respiratory syndrome coronavirus 2 epidemic><Severe acute respiratory syndrome coronavirus 2 pandemic><Shock Lung><Signal Transduction><Signal Transduction Systems><Signaling><Signaling Molecule><Simvastatin><Site><Stiff lung><Stimulus><Structure><Suidae><Swine><Synvinolin><Therapeutic><Transcription Factor Proto-Oncogene><Transcription factor genes><Ubiquitin Protein Ligase><Ubiquitin-Protein Ligase Complexes><Ubiquitin-Protein Ligase E3><Variant><Variation><Vascular Permeabilities><Viral><biological signal transduction><cartilage link protein><cell adhesion protein><coronavirus disease 2019 crisis><coronavirus disease 2019 epidemic><coronavirus disease 2019 global health crisis><coronavirus disease 2019 global pandemic><coronavirus disease 2019 health crisis><coronavirus disease 2019 pandemic><coronavirus disease 2019 public health crisis><coronavirus disease crisis><coronavirus disease epidemic><coronavirus disease pandemic><coronavirus disease-19 global pandemic><coronavirus disease-19 pandemic><cortactin><denitration><disparity in health><driving><ems1 sequence (mammary tumor and squamous cell carcinoma-associated (p80/85 src substrate))><epigenetic regulation><health disparity><hemidesmosomal protein HD1><inhibitor><insight><integrin bound><intracellular skeleton><link protein><live cell image><live cell imaging><live cellular image><live cellular imaging><lung injury><mechanical respiratory assist><mechanically ventilated><mechanosensing><mechanotransduction><mortality><multiorgan failure><multiple organ system failure><mutant><nitration><novel><pathophysiology><paxillin><pharmacologic><pig model><piglet model><plasmalemma><plectin><polymorphism><porcine><porcine model><pp125FAK><pre-clinical><preclinical><promoter><promotor><pulmonary><pulmonary damage><pulmonary injury><pulmonary tissue damage><pulmonary tissue injury><recruit><response><restoration><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><social role><suid><swine model><three dimensional><transcription factor><translation strategy><translational approach><translational strategy><ubiquitin-protein ligase><vascular><wet lung>