title: Imaging pulmonary NF-kappaB activation and therapeutic effects of MLN120B and TDZD-8.
abstract: NF-κB activation is a critical signaling event in the inflammatory response and has been implicated in a number of pathological lung diseases. To enable the assessment of NF-κB activity in the lungs, we transfected a luciferase based NF-κB reporter into the lungs of mice or into Raw264.7 cells in culture. The transfected mice showed specific luciferase expression in the pulmonary tissues. Using these mouse models, we studied the kinetics of NF-κB activation following exposure to lipopolysaccharide (LPS). The Raw264.7 cells expressed a dose-dependent increase in luciferase following exposure to LPS and the NF-κB reporter mice expressed luciferase in the lungs following LPS challenge, establishing that bioluminescence imaging provides adequate sensitivity for tracking the NF-κB activation pathway. Interventions affecting the NF-κB pathway are promising clinical therapeutics, thus we further examined the effect of IKK-2 inhibition by MLN120B and glycogen synthase kinase 3 beta inhibition by TDZD-8 on NF-κB activation. Pre-treatment with either MLN120B or TDZD-8 attenuated NF-κB activation in the pulmonary tissues, which was accompanied with suppression of pro-inflammatory chemokine MIP-1ß and induction of anti-inflammatory cytokine IL-10. In summary, we have established an imaging based approach for non-invasive and longitudinal assessment of NF-κB activation and regulation during acute lung injury. This approach will potentiate further studies on NF-κB regulation under various inflammatory conditions.
authors.affiliation: Caliper Life Sciences, Alameda, California, United States of America.
authors.forename: Dan
authors.initials: D
authors.lastname: Ansaldi
authors.forename: Eldad A
authors.initials: EA
authors.lastname: Hod
authors.forename: Fabio
authors.initials: F
authors.lastname: Stellari
authors.forename: Jae-Beom
authors.initials: JB
authors.lastname: Kim
authors.forename: Ed
authors.initials: E
authors.lastname: Lim
authors.forename: Mark
authors.initials: M
authors.lastname: Roskey
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Francis
authors.forename: Rajendra
authors.initials: R
authors.lastname: Singh
authors.forename: Ning
authors.initials: N
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002460:Cell Line; D054407:Chemokine CCL4 / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors; D000071679:Glycogen Synthase Kinase 3 beta; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors; D016753:Interleukin-10 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000633:toxicity; D008168:Lung / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D013830:Thiadiazoles / Q000494:pharmacology*
chemical_list: C494356:4-benzyl-2-methyl-1,2,4-thiadiazolidine-3,5-dione; D000893:Anti-Inflammatory Agents; D054407:Chemokine CCL4; D004791:Enzyme Inhibitors; D008070:Lipopolysaccharides; D016328:NF-kappa B; D013830:Thiadiazoles; D016753:Interleukin-10; D000071679:Glycogen Synthase Kinase 3 beta; D051550:I-kappa B Kinase; C496562:Ikbkb protein, mouse; D038362:Glycogen Synthase Kinase 3