Generation of an efficiently secreted, cell penetrating NF-κB inhibitor

Koutsokeras, Apostolos, Purkayastha, Nirupam, Rigby, Anne, Subang, Maria C, Sclanders, Michelle, Vessillier, Sandrine, Mullen, Lisa, Chernajovsky, Yuti and Gould, David (2014) Generation of an efficiently secreted, cell penetrating NF-κB inhibitor. FASEB Journal, 28 (1). pp. 373-381. ISSN 0892-6638

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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.

Item Type: Article
Schools and Departments: Brighton and Sussex Medical School > Clinical and Experimental Medicine
Subjects: R Medicine
Depositing User: Gemma Hamilton
Date Deposited: 16 Nov 2018 14:43
Last Modified: 16 Nov 2018 14:43

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