Prevention of NF-kappaB activation in vivo by a cell-permeable NF-kappaB inhibitor peptide.
The NF-kappaB/Rel transcription factor family plays a central role in coordinating the expression of a variety of genes that regulate stress responses, immune cell activation, apoptosis, proliferation, differentiation, and oncogenic transformation. Interventions that target the NF-kappaB pathway may be therapeutic for a variety of pathologies, especially immune/inflammatory diseases. Using membrane translocating sequence (MTS) technology, we developed a cell-permeable dominant inhibitor of NF-kappaB activation, termed IkappaBalpha-(DeltaN)-MTS. This molecule contains a 12-amino acid MTS motif attached to the COOH-terminal region of a nondegradable inhibitor protein [IkappaBalpha-(DeltaN)]. The recombinant protein enters cells and localizes in the cytoplasm. Delivery of the IkappaBalpha-(DeltaN)-MTS to cell lines and primary cells inhibited nuclear translocation of NF-kappaB proteins induced by cell activation. The protein also effectively inhibited NF-kappaB activation in vivo in two different animal models: NF-kappaB activation in response to skin wounding in mice and NF-kappaB activation in lungs after endotoxin treatment in sheep. Inhibition of NF-kappaB by the IkappaBalpha-(DeltaN)-MTS in the endotoxin model attenuated physiological responses to endotoxemia. These data demonstrate that activation of NF-kappaB can be inhibited using a recombinant protein designed to penetrate into cells. This technology may provide a new approach to NF-kappaB pathway-targeted therapies.
Mora
Ana L
AL
Division of Pulmonary, Allergy, and Critical Care Medicine, Emory University School of Medicine, Atlanta, GA 30322, USA.
LaVoy
John
J
McKean
Martha
M
Stecenko
Arlene
A
Brigham
Kenneth L
KL
Parker
Richard
R
Rojas
Mauricio
M
D000818:Animals; D002455:Cell Division / Q000276:immunology; D002463:Cell Membrane Permeability; D016503:Drug Delivery Systems; D019446:Endotoxemia / Q000188:drug therapy / Q000276:immunology; D015870:Gene Expression; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics / Q000493:pharmacokinetics*; D008171:Lung Diseases / Q000188:drug therapy* / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D041681:NIH 3T3 Cells; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000493:pharmacokinetics*; D012756:Sheep; D013601:T-Lymphocytes / Q000187:drug effects; D013950:Thymus Gland / Q000166:cytology
D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606204:NFKBIA protein, human; C000606205:Nfkbia protein, mouse; D011993:Recombinant Fusion Proteins; D000072000:NF-KappaB Inhibitor alpha