Premium
Generation of an efficiently secreted, cell penetrating NF‐κB inhibitor
Author(s) -
Koutsokeras Apostolos,
Purkayastha Nirupam,
Rigby Anne,
Subang Maria C.,
Sclanders Michelle,
Vessillier Sandrine,
Mullen Lisa,
Chernajovsky Yuti,
Gould David
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.13-236570
Subject(s) - secretion , fusion protein , microbiology and biotechnology , paracrine signalling , intracellular , transcription factor , secretory protein , nf κb , chemistry , matrix metalloproteinase , cell culture , signal transduction , biology , gene , recombinant dna , receptor , biochemistry , genetics
Gene therapy is a powerful approach to treat disease locally. However, if the therapeutic target is intracellular, the therapeutic will be effective only in the cells where the therapeutic gene is delivered. We have engineered a fusion protein containing an intracellular inhibitor of the transcription factor NF‐κB pathway that can be effectively secreted from producing cells. This fusion protein is cleaved extracellularly by metalloproteinases allowing release of a protein transduction domain (PTD) linked to the NF‐κB inhibitor for translocation into neighboring cells. We show that engineered molecules can be efficiently secreted (>80%); are cleaved with matrix metalloprotease‐1; inhibit NF‐κB driven transcription in a biological assay with a human reporter cell line; and display significant inhibition in mouse paw inflammation models when delivered by lentivirus or secreting cells. No inhibition of NF‐κB transcription or therapeutic effect was seen using molecules devoid of the PTD and NF‐κB inhibitory domains. By creating a fusion protein with an endogenous secretion partner, we demonstrate a novel approach to efficiently secrete PTD‐containing protein domains, overcoming previous limitations, and allowing for potent paracrine effects.—Koutsokeras, A., Purkayastha, N., Rigby, A., Subang, M. C., Sclanders, M., Vessillier, S., Mullen, L., Chernajovsky, Y., Gould, D. Generation of an efficiently secreted, cell penetrating NF‐κB inhibitor. FASEB J . 28, 373–381 (2014). www.fasebj.org