Abstract
NF-κB is a family of transcription factors regulated through two distinct signaling cascades, the classical and the Noncanonical NF-κB pathways. Noncanonical NF-κB plays important roles in the immune system, as it is necessary for lymphoid organogenesis and B-cell survival and differentiation, as well as osteoclastogenesis. In the last few years, there has been an increased number of studies focusing on both identifying the upstream events that regulate the noncanonical NF-κB pathway as well as determining the physiological roles of noncanonical NF-κB in normal and disease pathologies, such as cancer and autoimmune diseases. Dysregulation of noncanonical NF-κB has now been associated with the pathogenesis of several types of lymphomas and autoimmune diseases and is believed to contribute to chronic inflammatory diseases, including ulcerative colitis. These studies suggest that targeting the Noncanonical pathway, similar to classical NF-κB, may have some therapeutic potential in the future; however, there is still quite a bit about the regulation of the noncanonical signaling that remains to be defined. In this chapter we describe the use of HUVEC, as an in vitro model for examining noncanonical NF-κB signaling in response to different stimuli. We demonstrate two different methods to measure noncanonical NF-κB activation: the processing of p100 to p52, and noncanonical NF-κB-dependent gene expression of CXCL12. The first method examines a key regulatory requirement for noncanonical NF-κB activation, by which p100 undergoes proteolytic cleavage to relieve the inhibition of NF-κB dimers for nuclear translocation and activation of gene transcription. The latter demonstrates the downstream effects of activated noncanonical NF-κB in response to stimuli.
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References
Hayden MS, Ghosh S (2004) Signaling to NF-kappaB. Genes Dev 18:2195–2224
Dejardin E, Droin NM, Delhase M et al (2002) The lymphotoxin-β receptor induces different patterns of gene expression via two NF-kappaB pathways. Immunity 17:525–535
Coope HJ, Atkinson PG, Huhse B et al (2002) CD40 regulates the processing of NF-kappaB2 p100 to p52. EMBO J 15:5375–5385
Claudio E, Brown K, Park S et al (2002) BAFF-induced NEMO-independent processing of NF-kappaB2 in maturing B cells. Nat Immunol 3:958–965
Novack DV, Yin L, Hagen-Stapleton A et al (2003) The IκB function of NF-κB2 p100 controls stimulated osteoclastogenesis. J Exp Med 198:771–781
Sun S-C (2011) Non-canonical NF-κB signaling pathway. Cell Res 21:71–85
Courtois G, Gilmore TD (2006) Mutations in the NF-κB signaling pathway: implications for human disease. Oncogene 25:6831–6843
Karin M (2006) Nuclear factor-κB in cancer development and progression. Nature 441:431–436
Tak PP, Firestein GS (2001) NF-κB: a key role in inflammatory disease. J Clin Invest 107:7–11
Baker RG, Hayden MS, Ghosh S (2011) NF-κB, inflammation and metabolic disease. Cell Metab 13:11–22
Dejardin E (2006) The alternative NF-κB pathway from biochemistry to biology: pitfalls and promises for future drug development. Biochem Pharmacol 72:1161–1179
Ranzani B, Reichardt AD, Cheng G (2011) Non-canonical NF-κB signaling activation and regulation: principles and perspectives. Immunol Rev 244:44–54
Brown KD, Claudio E, Siebenlist U (2008) The roles of the classical and alternative nuclear factor-κB pathway: potential implications for autoimmunity and rheumatoid arthritis. Arthritis Res Ther 10:212
Sitia S, Tomasoni L, Atzeni F et al (2010) From endothelial dysfunction to atherosclerosis. Autoimmun Rev 9:830–834
Solan NJ, Miyoshi H, Carmona EM et al (2002) RelB cellular regulation and transcriptional activity are regulated by p100. J Biol Chem 277:1405–1418
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McCorkell, K.A., May, M.J. (2015). Noncanonical NF-κB Activation and SDF-1 Expression in Human Endothelial Cells. In: May, M. (eds) NF-kappa B. Methods in Molecular Biology, vol 1280. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2422-6_9
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DOI: https://doi.org/10.1007/978-1-4939-2422-6_9
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