Endothelial Biology Core:
The Endothelial Cell Biology Core provides in vitro studies of pulmonary endothelial cell function. The research goals focus on identifying signaling pathways and post-translational modifications that result in cytoskeletal rearrangements and changes in endothelial barrier regulation. The Core uses a wide variety of biochemical and proteomic techniques, such as Western blots, immunofluorescence, immunoprecipitation, yeast two-hybrid, ELISA, RNA interference, and mass spectrometry, as well as emerging techniques such as atomic force microscopy, microRNA, and nanoparticle encapsulation for targeted delivery of drugs/agents. In particular, the Core specializes in measuring endothelial barrier permeability using two well-established assays, FITC-dextran clearance assay and Electrical Cell-substrate Impedance Sensing (ECIS) assay (www.appliedbiophysics.com). The Core also specializes in in vitro studies of ventilator-induced lung injury (VILI) by cyclic stretching cultured lung endothelial cells using Flexcell apparatus (www.flexcellint.com).
Animal & Transgenic Core:
The Animal and Transgenic Core provides the capacity to conduct in vivo studies of endothelial barrier function using preclinical models (mice, rats, dogs) of acute lung injury (ALI) or acute respiratory distress syndrome (ARDS), asthma, pulmonary hypertension, congestive heart failure, radiation pneumonitis, and lung cancer. The research focus is to understand the cellular and molecular basis of lung vascular dysfunction including vascular remodeling, endothelial barrier permeability in response to inflammatory agonists, such as LPS and airborne particulate matter, as well as ventilator-induced lung injuries (VILI). The Animal Core employs state of-the-art evaluation of pulmonary pathophysiology, interventions, imaging, data analysis, and interpretation, in order to provide insight into the efficacy and mechanisms of clinically relevant management approaches and to facilitate the translation of basic research to the clinical arena. Potential therapeutic treatments include the delivery of drugs such as S1P, FTY720, and simvastatin, as well as the delivery of targeted protein silencing using nanoparticle-encapsulated siRNA. In addition, the Transgenic Core generates novel transgenic mice as directed by the needs of the studies and cares for the genetically engineered mice on a daily basis. The transgenic mice collection includes knockout mice (nmMLCK, TLR4), heterozygous mice (PBEF), endothelial-specific overexpressing mice (nmMLCK-EC), and endothelial-specific overexpressing mice crossed with knockout mice (MLCK-EC/MLCK-KO combo).
The Genomics Core provides expert assessment of in silico studies of inflammatory lung injury models with identifying genetic variations among different populations and species that link to lung disease susceptibility and severity. The Core utilizes techniques such as real-time RT-PCR, microarrays, SNP discovery, and single loci association and haplotype analysis. The Core has identified several candidate genes for acute respiratory distress syndrome (ARDS) particularly using orthologous gene-expression profiling in multi-species models including samples from pathophysiological-stretched cultured human endothelial cells, murine and canine lung tissues from in vivo lung injury models, as well as human patient samples. The Core also focuses on identification of SNPs and haplotypes in the candidate genes from Caucasian and African-American cohorts, utilizing knowledge that African-American patients are genetically predisposed to be susceptible to developing ARDS. Through case-control association studies, the Core has identified genetic polymorphisms associated with susceptibility to and severity of ARDS and asthma. The Genomics Core has dynamic interactions with both the Animal and Endothelial Biology Cores as the identification of candidate genes provide novel targets to silence or inhibit for in vitro experiments as well as to for the generation of transgenic mice for in vivo experimentations.