Molecular pharmacology of lung with focus on effects of oxidative and nitrosative stress on pulmonary endothelium and smooth muscle.

Description of projects available to graduate students:
Dr. Pitt’s laboratory efforts are directed towards original studies on the molecular and cellular biology of lung. To date, this work has focused primarily on the role of oxidants and nitric oxide in affecting pulmonary endothelial and vascular smooth muscle cell function. Isolated primary cell cultures, genetically modified murine models and somatic gene transfer to lung have been used as model systems to identify the role of partially reduced oxygen and nitrogen species in the response of the lung to stress and injury.

Recent efforts have been directed towards understanding the role of S-nitrosylation of the metal binding intracellular protein, metallothoinein, on zinc homeostasis and endothelial cell function. These studies are aided by a number of microspectrofluorometric approaches including a green fluorescent protein modified chimera of metallothionein, suitable for fluorescence resonance energy transfer. This chimera contains an enhanced cyan fluorescent protein (ECYP) that serves as a donor for an enhanced yellow flourescence protein (EYFP) that are fused to the N- and C-termini respectively of human metallothionein IIa (hMTIIa). The result of these live cell fluorescence image studies has identified a unique role for metallothionein in NO mediated zinc release. NO mediated changes in labile zinc appear to be inhibitory in proapoptotic conditions in pulmonary endothelial cells contributing to the notion that S-nitrosylation of zinc sulfur clusters is an important component of NO signaling.

Techniques graduate student will learn:
isolation and culture of pulmonary endothelial cells; cell and organ imaging; somatic gene tranfer; transgenic mice; effect of oxidants and nitric oxide on perfused lungs and intact animals.