1. Lung Injury and nucleotides. (Dr. Christelle Douillet, Dr. Preston Rich)
We developed a model of ventilated rats, in which different ventilatory protocols are applied, before assessing lung and systemic injuries. Using drugs, we also manipulate the extracellular nucleotide economy in the lungs or vascular compartment and identify the effects of nucleotides on ventilated animals. We determine lung injury and multiple organ failure by different methods, including histology, biochemistry, molecular biology (i.e. measuring gene expression by real-time PCR) and evaluating edema, inflammation, fibrosis, apoptosis etc. We determine if these outcomes are dependent on the nucleotide economy.
We use a model of ventilated mice, which are wild-type or purinoceptor knock-outs. We evaluate the role of nucleotides and purinoceptors by comparing lung injury and MOF (as evaluated in sub-project 1.1) in the different types of mice.
We use cell culture models of airway epithelia (tracheal, bronchial, and small airways), pneumocytes, and lung smooth muscle cells to determine the effects of nucleotides on lung cell physiology. 3 types of cellular function are studied: cytokine expression and secretion, extracellular matrix regulation, and apoptosis. We determine dose and time-responses curves of several nucleotides, agonist profiles, the type of receptor involved, and the regulation between expression/translation/secretion, etc. Particular emphasis is placed on defining the intracellular signaling pathways involved in those nucleotide effects. Cells are also submitted to strain and pressure (mimicking ventilation) by Flexercell Strain Unit, in presence or not of nucleotide economy modifiers, and cellular function (as above) studied.
In patients with lung injury (ARDS, Acute Lung Injury, pulmonary contusion, pneumonia, or inhalation injury. IRB approved), we will assess broncho-alveolar lavage nucleotides and compare nucleotide concentrations and profiles with the different types of lung injury. We will also correlate the nucleotide data with other outcomes (cell death, inflammation, edema markers, purinoceptor expression in lavage cells, etc).
2. Aortic aneurysm and nucleotides
We use human aortic smooth muscle cells in culture to determine the effect of nucleotides on the regulation on extracellular matrix. We determine dose and time-response curves of several nucleotides, agonist profiles, the type of receptor involved, signaling pathways, and expression/translation/secretion regulations of matrix metalloproteinases and their tissue inhibitors (TIMPs).
We collect aortic tissue (IRB approved) from patient aneurysms. We determine tissue nucleotide concentrations and profiles, purinoceptor content and expression, extracellular matrix, metalloproteinase and TIMP content and expression. We will compare these data to those from normal aortic tissue (collected during CABG) to determine if nucleotides are involved in the regulation of extracellular matrix in vivo. .
3. Traumatic brain and spinal cord injury and nucleotides
Human CSF samples will be collected (IRB approved) in order to elucidate the presence and role of extracellular nucleotides in traumatic brain and spinal cord injuries. The nucleotide profiles will then be correlated with markers of cellular injury, oxidative damage, inflammation, and EPO production. In addition, a cell culture model will be used to further establish the role of extracellular nucleotides in the expression EPO in brain and spinal cord injury.
4. Evaluation of traumatic injuries with highly sensitive infrared thermography.
In animal models, we assess highly sensitive infrared thermography as a diagnostic tool for detecting open- or closed-chest pneumothorax, hemothorax, open- or closed-abdomen hemoperitoneum, and stroke.
- Tissue Culture Facility (Lineberger Comprehensive Cancer Center)
- Immunotechnology Core (Center for GastroIntestinal Biology and Disease)
- Microscopy Services Laboratory Core Facility
- Histopathology Core facility
- General Clinical Research Center
- UNC-CH Center for Bioinformatics
- Flow Cytometry Core Facility
- Cystic Fibrosis/Pulmonary Research and Treatment Center
- Dr. Richard C. Boucher (Dept of Medicine)
- Dr. Eduardo Lazarowski (Dept of Medicine)
- Dr Wanda O’Neal (Dept of Medicine)
- Dr Stephen Tilley (Dept of Medicine)
- Dr. Larry Katz (Dept of Emergency Medicine)
- Dr. Cam Patterson (Dept of Medicne, Carolina Center for Cardiovascular Research)
- Dr. Craig Selzman (Dept of Surgery, Division of Cardiothoracic Surgery)
- Dr. Blair Keagy (Dept of Surgery, Division of Vascular Surgery)
- Dr. Mark Farber (Dept of Surgery, Division of Vascular Surgery)
- Dr. Matt Ewend (Dept of Surgery, Division of Neurosurgery)
- Dr. Eldad Hadar (Dept of Surgery, Division of Neurosurgery)