Document text
Principal Investigator: Joe G. N. Garcia
Organization: UNIVERSITY OF FLORIDA
Fiscal Year: 2024
Award: $297,844
Funding agency: National Heart Lung and Blood Institute
ABSTRACT:
The global COVID-19 pandemic has dramatically highlighted the critical role of lung vascular inflammation and
multi-organ endothelial cell (EC) permeability in ARDS mortality and the unprecedented COVID-19-ARDS vas-
cular endotype (1). This A1 Project #2 application remains focused on the critical role of eNAMPT (extracellular
nicotinamide phosphoribosyltransferase) in driving lung vascular inflammation and multi-organ endothelial
cell (EC) permeability, events that are central to increasing ARDS mortality. We initially identified eNAMPT as a
novel ARDS and ventilator-induced lung injury (VILI) therapeutic target utilizing genomic–intensive approaches
and cellular and preclinical studies of excessive mechanical stress/VILI. We showed eNAMPT is a novel ARDS
biomarker with plasma eNAMPT levels increasing in response to viral/bacterial infection and exposure to me-
chanical ventilation. Importantly, utilizing conditional EC–specific Nampt KO mice, we have recently shown that
EC contributions to ARDS pathobiology via eNAMPT secretion into the circulation, thereby driving pre-
clinical ARDS inflammatory lung injury and severity. eNAMPT produces these injurious effects by functioning as
a damage-associated molecular pattern protein (DAMP) and master regulator of evolutionarily-conserved inflam-
matory cascades via novel ligation of the Toll–like receptor 4 (TLR4). Our exciting data in mouse, rat and porcine
ARDS/VILI models have validated the efficacy of the eNAMPT-neutralizing humanized mAb (ALT-100) in re-
ducing eNAMPT- and LPS-induced TLR4 activation, NFκB-driven cytokine production, lung permeability and
inflammatory lung injury. To further interrogate eNAMPT as an ARDS therapeutic target, SA #1 will extend prior
studies which showed ROS-generating ARDS stimuli (hypoxia, hyperoxia, mechanical stress, cytokines) to in-
duce NAMPT expression and characterize the role of three key transcription factors (hypoxia-inducible factors
HIF1a/2a, NRF2), NAMPT/TLR4 promoter SNPs, and DNA methylation in genetic/epigenetic regulation of
NAMPT/TLR4 promoter activities. SA #2 will mechanistically explore novel regulation of TLR4- and mechanical
stress-stimulated eNAMPT secretion, a key event to initiation of inflammatory cascade activation, via extracellu-
lar vesicle formation, inflammasome activation, and ABC transporters. Utilizing Core B's CRISPR/Cas9- gener-
ated EC lines, SA #3 will dissect the structure/function mechanisms involved in eNAMPT-TLR4 binding and
increases in EC permeability with specific focus on MAP kinase effector p90rsk, Akt1 nitration, and UCHL1
activity in EC cytoskeletal-driven barrier dysfunction. Finally, utilizing Core C expertise, SA #4 will optimize eN-
AMPT ALT-100 mAb dosing, define the therapeutic window for ALT-100 delivery, and define potential ALT-100
mAb synergy with other PPG therapeutic modalities utilizing preclinical rat and porcine ARDS/VILI models. This
high integration of Project #2 with each PPG Project will elucidate EC secretion of eNAMPT and eNAMPT/TLR4
participation in ARDS/VILI pathobiology, and will accelerate ALT-100 mAb as an actionable strategy to attenuate
inflammatory EC permeability and reduce ARDS/VILI mortality.
Terms: <ABC Transport Protein><ABC Transporter Protein><ABC Transporters><ARDS><ATP-Binding Cassette Transporters><Acceleration><Acute Respiratory Distress><Acute Respiratory Distress Syndrome><Adult ARDS><Adult RDS><Adult Respiratory Distress Syndrome><Attenuated><Automobile Driving><Bacterial Infections><Basal Transcription Factor><Basal transcription factor genes><Binding><Biological Markers><Blood Plasma><Blood Vessels><COVID associated ARDS><COVID associated acute respiratory distress syndrome><COVID crisis><COVID epidemic><COVID induced ARDS><COVID induced acute respiratory distress syndrome><COVID pandemic><COVID related ARDS><COVID related acute respiratory distress syndrome><COVID-19 associated ARDS><COVID-19 associated acute respiratory distress syndrome><COVID-19 crisis><COVID-19 epidemic><COVID-19 era><COVID-19 global health crisis><COVID-19 global pandemic><COVID-19 health crisis><COVID-19 induced ARDS><COVID-19 induced acute respiratory distress syndrome><COVID-19 infection><COVID-19 pandemic><COVID-19 period><COVID-19 public health crisis><COVID-19 related ARDS><COVID-19 related acute respiratory distress syndrome><COVID-19 virus infection><COVID-19 years><COVID-19/ARDS><COVID-19/acute respiratory distress syndrome><COVID/ARDS><COVID/acute respiratory distress syndrome><COVID19 associated ARDS><COVID19 associated acute respiratory distress syndrome><COVID19 induced ARDS><COVID19 induced acute respiratory distress syndrome><COVID19 infection><COVID19 related ARDS><COVID19 related acute respiratory distress syndrome><COVID19/ARDS><COVID19/acute respiratory distress syndrome><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cas nuclease technology><Cell Communication and Signaling><Cell Line><Cell Signaling><Cell secretion><CellLine><Cellular Matrix><Cellular Secretion><Circulation><Clinical Treatment Moab><Closure by Ligation><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Common Rat Strains><Cytoskeletal System><Cytoskeleton><DNA Methylation><Da Nang Lung><Data><Deubiquitination><Dimerization><Dose><Dysfunction><Endothelial Cells><Endothelium><Event><Exposure to><FK506 Binding Protein 12-Rapamycin Associated Protein 1><FKBP12 Rapamycin Complex Associated Protein 1><FRAP1><FRAP1 gene><FRAP2><Family suidae><Functional disorder><General Transcription Factor Gene><General Transcription Factors><Genetic><Genomics><HIF 1 alpha><HIF-1alpha><HIF1-Alpha><HIF1A><HIF1A gene><HIF1α><Homolog of Drosophila TOLL><Hyperoxia><Hypoxia><Hypoxia Inducible Factor><Hypoxic><Inflammasome><Inflammatory><Injury><Intracellular Communication and Signaling><KO mice><Knock-out Mice><Knockout Mice><Ligation><Lung><Lung Respiratory System><Lung damage><MAP kinase><MOP1><Mechanical Stress><Mechanical ventilation><Mechanistic Target of Rapamycin><Mediating><Mice><Mice Mammals><Mitogen-Activated Protein Kinases><Modality><Modeling><Molecular><Molecular Interaction><Monoclonal Antibodies><Murine><Mus><NMN pyrophosphorylase><Null Mouse><Organ><Oxygen Deficiency><PARK5><Parkinson disease 5 gene><Pathway interactions><Pattern><Permeability><Physiopathology><Pigs><Plasma><Plasma Serum><Predisposition><Production><Protein Dimerization><Proteins><RAFT1><Rat><Rats Mammals><Rattus><Regulation><Reticuloendothelial System, Serum, Plasma><Role><SARS-CoV-2 associated ARDS><SARS-CoV-2 associated acute respiratory distress syndrome><SARS-CoV-2 epidemic><SARS-CoV-2 global health crisis><SARS-CoV-2 global pandemic><SARS-CoV-2 induced ARDS><SARS-CoV-2 induced acute respiratory distress syndrome><SARS-CoV-2 infection><SARS-CoV-2 pandemic><SARS-CoV-2 related ARDS><SARS-CoV-2 related acute respiratory distress syndrome><SARS-CoV-2/ARDS><SARS-CoV-2/acute respiratory distress syndrome><SARS-CoV2 infection><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 infection><Severe acute respiratory syndrome coronavirus 2 pandemic><Severities><Shock Lung><Short interfering RNA><Signal Transduction><Signal Transduction Systems><Signaling><Small Interfering RNA><Stiff lung><Stimulus><Strains Cell Lines><Structure><Suidae><Susceptibility><Swine><TLR4><TLR4 gene><Therapeutic><Toll Homologue><Transcription Factor Proto-Oncogene><Transcription factor genes><UCHL1><UCHL1 gene><Ubiquitilation><Ubiquitination><Ubiquitinoylation><Ventilator><Ventilator-induced lung injury><Vesicle><Viral><attenuate><attenuates><bacteria infection><bacterial disease><bio-markers><biologic marker><biological signal transduction><biomarker><coronavirus disease 2019 associated ARDS><coronavirus disease 2019 associated acute respiratory distress syndrome><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 induced ARDS><coronavirus disease 2019 induced acute respiratory distress syndrome><coronavirus disease 2019 infection><coronavirus disease 2019 pandemic><coronavirus disease 2019 public health crisis><coronavirus disease 2019 related ARDS><coronavirus disease 2019 related acute respiratory distress syndrome><coronavirus disease 2019/ARDS><coronavirus disease 2019/acute respiratory distress syndrome><coronavirus disease associated ARDS><coronavirus disease associated acute respiratory distress syndrome><coronavirus disease crisis><coronavirus disease epidemic><coronavirus disease induced ARDS><coronavirus disease induced acute respiratory distress syndrome><coronavirus disease pandemic><coronavirus disease related ARDS><coronavirus disease related acute respiratory distress syndrome><coronavirus disease-19 global pandemic><coronavirus disease-19 pandemic><coronavirus disease/ARDS><coronavirus disease/acute respiratory distress syndrome><cultured cell line><cytokine><de-ubiquitinase><de-ubiquitinating enzyme><denitration><driving><efficacy validation><epigenetic regulation><extracellular><extracellular vesicles><hyperoxygenation><infected with COVID-19><infected with COVID19><infected with SARS-CoV-2><infected with SARS-CoV2><infected with coronavirus disease 2019><infected with severe acute respiratory syndrome coronavirus 2><injuries><intracellular skeleton><lung injury><lung vascular inflammation><mAbs><mTOR><mammalian target of rapamycin><mechanical respiratory assist><mechanically ventilated><monoclonal Abs><mortality><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><nicotinamide phosphoribosyltransferase><nitration><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><pathophysiology><pathway><pig model><piglet model><porcine><porcine model><pre-clinical><pre-clinical study><preclinical><preclinical study><promoter><promotor><pulmonary><pulmonary damage><pulmonary injury><pulmonary tissue damage><pulmonary tissue injury><pulmonary vascular inflammation><response><severe acute respiratory syndrome coronavirus 2 associated ARDS><severe acute respiratory syndrome coronavirus 2 associated acute respiratory distress syndrome><severe acute respiratory syndrome coronavirus 2 global health crisis><severe acute respiratory syndrome coronavirus 2 global pandemic><severe acute respiratory syndrome coronavirus 2 induced ARDS><severe acute respiratory syndrome coronavirus 2 induced acute respiratory distress syndrome><severe acute respiratory syndrome coronavirus 2 related ARDS><severe acute respiratory syndrome coronavirus 2 related acute respiratory distress syndrome><severe acute respiratory syndrome coronavirus 2/ARDS><severe acute respiratory syndrome coronavirus 2/acute respiratory distress syndrome><siRNA><social role><suid><swine model><synergism><therapeutic target><toll-like receptor 4><transcription factor><ubiquination><ubiquitin conjugation><ubiquitin isopeptidase><ubiquitin-specific isopeptidase><validate efficacy><vascular><ventilation induced lung injury><ventilator associated lung injury><wet lung>