Acute respiratory failure (ARF) requiring mechanical ventilation (MV) affects as many as 150,000 people per year in the United States alone, resulting in a greater incidence than either colon cancer or AIDS. Associated mortality rates are staggering (45-95%), and costs are estimated in the billions of dollars. Although potentially lifesaving, evidence has implicated MV in inducing lung injury (ventilator-induced lung injury, VILI) and contributing to multiple system organ dysfunction (MSOD). Despite a growing understanding of the complex pathophysiologic events associated with MV, ARF, and MSOD, the causes of VILI remain unclear. It is known that nucleotides are 1) released into the extracellular environment in response to shear stress, and 2) regulate numerous cellular processes, including cytokine release, monocyte and fibroblast proliferation, iNOS, adhesion molecule expression, ion transport, platelet degranulation, and apoptosis. Although these processes have individually been implicated in VILI, the central role of nucleotides in orchestrating these pathways has not been studied. We have observed that purine nucleotides are released during large-volume MV; furthermore, pharmacological manipulations of nucleotide levels implicate them in VILI- associated inflammation and alveolar fluid metabolism. We hypothesize that nucleotides play a central role in VILI, and to address this we propose three specific aims: 1) measure the basal and stimulated nucleotide concentrations in the alveolar and vascular compartments in response to selected airway stimuli, 2) determine the relationship of nucleotides to pulmonary and systemic inflammation during MV, and 3) examine the effect of nucleotides on transalveolar fluid dynamics. The long-term goal of this project will be to further our understanding of the mechanisms responsible for VILI. This proposal is designed for long-term scientific success, and benefits greatly from the intense supervision and oversight provided by an established investigator with a proven track-record of successful mentorship. A detailed 5-year plan is outlined, which frames innovative scientific research, appropriate didactic coursework, technical workshops, direct laboratory supervision, and ethical training in an environment with a strong academic tradition and commitment to training clinical scientists. The proposed research and career development plan will prepare the candidate to be an independent scientific investigator in the field of lung injury.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Clinical Investigator Award (CIA) (K08)
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Special Emphasis Panel (ZHL1-CSR-M (F2))
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Colombini-Hatch, Sandra
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University of North Carolina Chapel Hill
Schools of Medicine
Chapel Hill
United States
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Milano, Peter M; Douillet, Christelle D; Riesenman, Paul J et al. (2008) Intestinal ischemia-reperfusion injury alters purinergic receptor expression in clinically relevant extraintestinal organs. J Surg Res 145:272-8
Carson, Shannon S; Kress, John P; Rodgers, Jo Ellen et al. (2006) A randomized trial of intermittent lorazepam versus propofol with daily interruption in mechanically ventilated patients. Crit Care Med 34:1326-32
Robinson 3rd, William P; Douillet, Christelle D; Milano, Peter M et al. (2006) ATP stimulates MMP-2 release from human aortic smooth muscle cells via JNK signaling pathway. Am J Physiol Heart Circ Physiol 290:H1988-96
Douillet, Christelle D; Robinson 3rd, William P; Milano, Peter M et al. (2006) Nucleotides induce IL-6 release from human airway epithelia via P2Y2 and p38 MAPK-dependent pathways. Am J Physiol Lung Cell Mol Physiol 291:L734-46
Douillet, Christelle D; Robinson 3rd, William P; Zarzaur, Ben L et al. (2005) Mechanical ventilation alters airway nucleotides and purinoceptors in lung and extrapulmonary organs. Am J Respir Cell Mol Biol 32:52-8
Douillet, Christelle D; Suy, Sihong; Zarzaur, Ben L et al. (2005) Measurement of free and bound fractions of extracellular ATP in biological solutions using bioluminescence. Luminescence 20:435-41
Rich, Preston B; Douillet, Christelle D; Hurd, Harry et al. (2003) Effect of ventilatory rate on airway cytokine levels and lung injury. J Surg Res 113:139-45
Rich, Preston B; Douillet, Christelle D; Mahler, Simon A et al. (2003) Adenosine triphosphate is released during injurious mechanical ventilation and contributes to lung edema. J Trauma 55:290-7