Reactive oxygen species generation following cytokine stimulation.
The Lamb lab is focused on defining the biological impact of superoxide produced by NADPH oxidase 1(Nox1) following cytokine (TNFa, IL-1b, TLR4) receptor activation in vascular smooth muscle cells. We are particularly interested in the mechanisms by which anion conductances, including the ClC-3 Cl-/H+anti-porter and the swelling-induced anion channel (IClswell or LRRC8), support superoxide production within “signaling” endosomes that form in response to cytokine receptor endocytosis. We employ a multi-faceted approach to understanding all aspects of this system including; 1) the biophysical properties of ClC-3 and LRRC8 ion currents, 2) the nature of the intracellular vesicles where Nox1 generates superoxide, and 3) the regulatory influences (particularly MAP kinases) that control this signaling pathway. Our long term goal is to develop selective anti-inflammatory agents that can impair cytokine signaling and control vascular inflammation via inhibition of anion transport. Techniques employed include patch-clamp recording of ion currents, localization and quantitation of superoxide production, confocal microscopic imaging of labelled ion channels and intracellular vesicular proteins as well as a wide variety of basic molecular biologic assays.