Benjamin Gaston, MD
Principal Investigator, TITUS Group Project 1
Chief, Division of Pediatric Pulmonology, and Allergy/Immunology
Department of Pediatrics
Case Western Reserve University
UH Rainbow Babies and Children’s Hospital
My laboratory studies cysteine modifications downstream of nitric oxide synthase (NOS) activation. These are relevant to cystic fibrosis (CF), asthma, pulmonary arterial hypertension and apnea. I have nearly two decades’ experience with S-nitrosothiol assays, and state-of-the-art technology for detecting cysteine modifications downstream of NOS activation. I also have extensive experience studying NOS in cell and molecular biology.
For nearly two decades, Dr. Gaston has studied S-nitrosothiol signaling in the airway epithelium. He has discovered that S-nitrosoglutathione (GSNO) concentrations are depleted in the asthmatic airway epithelium both in animal models and in humans (5,17). He and his coworkers, including Drs. Palmer and Zaman, also discovered that GSNO and other S-nitrosylating agents increase expression, maturation and function of DF508 cystic fibrosis (CF) transmembrane regulatory protein, potentially serving as a class of agents that could be corrector therapies for CF (18). Both of these observations have led to substantial translational projects regarding understanding the pathophysiology and treatment of asthma and CF.
More recently, Dr. Gaston’s group has collaborated closely with the laboratories of Drs. Palmer, Lewis and Periasamy to study the intracellular trafficking of S-nitrosothiols and the cellular localization of their actions and metabolism in airway epithelial cells. In particular, he, Dr. Lewis and Dr. Palmer have characterized the subcellular location of specific S-nitrosothiols, and of S-nitrosothiol metabolic enzymes. While these data reveal a novel signaling system of substantial potential relevance to respiratory therapeutics, many questions remain unanswered, particularly in the field of airway biology. For example, it is not known how NOS activation can target particular proteins for S-nitrosylation, nor where these reactions and/or formation of intermediate transnitrosation precursors are located in the airway epithelial cell.
To address these questions, Dr. Gaston will undertake three aims. He will test the hypotheses that: 1) specific proteins in normal human airway epithelial cells are S-nitrosylated by NOS; 2) protein S-nitrosylation occurs in specific subcellular locations in human airway epithelial cells; and 3) S-nitrosylation signaling is disordered in the human cystic fibrosis airway epithelium.
This project will involve extensive synergy with the Palmer and Lewis laboratories. It will extend observations, made previously in established cell culture lines, to primary pseudostratified columnar epithelial cell cultures from human subjects (Core B), and it will expand on observations regarding confocal colocalization of S-nitrosylated proteins and S-nitrosothiol metabolic proteins in epithelial cells (Core A). He will also, as overall PI, coordinate speakers and inter-institutional collaboration to establish the program as a center of excellence in S-nitrosothiol signaling in respiratory biology (Core C). At the conclusion of this project, we anticipate that we will have a detailed understanding the role of S-nitrosothiol localization, storage and transfer in regulating airway epithelial cell signaling reactions.