NF-κB inhibitor targeted to activated endothelium demonstrates a critical role of endothelial NF-κB in immune-mediated diseases.
Activation of the nuclear transcription factor κB (NF-κB) regulates the expression of inflammatory genes crucially involved in the pathogenesis of inflammatory diseases. NF-κB governs the expression of adhesion molecules that play a pivotal role in leukocyte-endothelium interactions. We uncovered the crucial role of NF-κB activation within endothelial cells in models of immune-mediated diseases using a "sneaking ligand construct" (SLC) selectively inhibiting NF-κB in the activated endothelium. The recombinant SLC1 consists of three modules: (i) an E-selectin targeting domain, (ii) a Pseudomonas exotoxin A translocation domain, and (iii) a NF-κB Essential Modifier-binding effector domain interfering with NF-κB activation. The E-selectin-specific SLC1 inhibited NF-κB by interfering with endothelial IκB kinase 2 activity in vitro and in vivo. In murine experimental peritonitis, the application of SLC1 drastically reduced the extravasation of inflammatory cells. Furthermore, SLC1 treatment significantly ameliorated the disease course in murine models of rheumatoid arthritis. Our data establish that endothelial NF-κB activation is critically involved in the pathogenesis of arthritis and can be selectively inhibited in a cell type- and activation stage-dependent manner by the SLC approach. Moreover, our strategy is applicable to delineating other pathogenic signaling pathways in a cell type-specific manner and enables selective targeting of distinct cell populations to improve effectiveness and risk-benefit ratios of therapeutic interventions.
Department of Rheumatology and Clinical Immunology and Centre of Chronic Immunodeficiency, University Medical Centre and University of Freiburg, 79106 Freiburg, Germany.
Bettina
B
Sehnert
Harald
H
Burkhardt
Johannes T
JT
Wessels
Agnes
A
Schröder
Michael J
MJ
May
Dietmar
D
Vestweber
Jochen
J
Zwerina
Klaus
K
Warnatz
Falk
F
Nimmerjahn
Georg
G
Schett
Stefan
S
Dübel
Reinhard Edmund
RE
Voll
D000818:Animals; D001168:Arthritis / Q000188:drug therapy* / Q000276:immunology*; D001426:Bacterial Proteins / Q000378:metabolism; D003001:Cloning, Molecular; D019040:E-Selectin / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D042783:Endothelial Cells / Q000187:drug effects / Q000276:immunology*; D004926:Escherichia coli; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology*; D051379:Mice; D016328:NF-kappa B / Q000037:antagonists & inhibitors*; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000494:pharmacology; D015398:Signal Transduction / Q000276:immunology
D001426:Bacterial Proteins; D019040:E-Selectin; D016328:NF-kappa B; D011993:Recombinant Fusion Proteins; C111749:pseudomonas exoprotein A protein, Pseudomonas aeruginosa
autoimmune disorders; cell targeting; inhibit inflammation; intracellular signaling; mouse models