Submitted on April 12, 2005
Accepted on June 6, 2005
Prevention of NF-
B Activation In vivo by a Cell Permeable NF-B Inhibitor Peptide
Ana L Mora1, John LaVoy2, Martha McKean2, Arlene Stecenko1, Kenneth L Brigham1, Richard Parker2, and Mauricio Rojas2*
1 Division of Pulmonary, Allergy and Critical Care Medicine, Emory University, Atlanta, GA, USA; Center for Translational Reseach in the Lung, Atlanta, GA, USA; Department of Medicine, Emory University School of Medicine, McKelvey Center for Lung Transplantation, Atlanta, GA, USA
2 Division of Pulmonary, Allergy and Critical Care Medicine, Emory University, Atlanta, GA, USA; Center for Translational Reseach in the Lung, Atlanta, GA, USA
* To whom correspondence should be addressed. E-mail: mrojas{at}emory.edu.
The NF-
B/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-B 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-B activation, termed IB
-(
N)-MTS. This molecule contains a 12 amino acid MTS motif attached to the carboxy terminal region of a non-degradable inhibitor protein (IB-[DELTA]]N)). The recombinant protein enters cells and localizes in the cytoplasm. Delivery of the IB-(N)-MTS to cell lines and primary cells inhibited nuclear translocation of NF-B proteins induced by cell activation. The protein also effectively inhibited NF-B activation in vivo in two different animal models: NF-B activation in response to skin wounding in mice and NF-B activation in lungs after endotoxin treatment in sheep. Inhibition of NF-B by the IB-(N)-MTS in the endotoxin model attenuated physiologic responses to endotoxemia. These data demonstrate that activation of NF-B can be inhibited using a recombinant protein designed to penetrate into cells. This technology may provide a new approach to NF-B pathway targeted therapies.
