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Generation of an efficiently secreted, cell penetrating NF-?B inhibitor
journal contribution
posted on 2023-06-09, 15:55 authored by Apostolos Koutsokeras, Nirupam Purkayastha, Anne Rigby, Maria C Subang, Michelle Sclanders, Sandrine Vessillier, Lisa MullenLisa Mullen, Yuti Chernajovsky, David GouldGene 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-?B 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-?B inhibitor for translocation into neighboring cells. We show that engineered molecules can be efficiently secreted (>80%); are cleaved with matrix metalloprotease-1; inhibit NF-?B 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-?B transcription or therapeutic effect was seen using molecules devoid of the PTD and NF-?B 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.
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Publication status
- Published
File Version
- Published version
Journal
FASEB JournalISSN
0892-6638Publisher
Federation of American Society of Experimental BiologyExternal DOI
Issue
1Volume
28Page range
373-381Department affiliated with
- Clinical and Experimental Medicine Publications
Full text available
- No
Peer reviewed?
- Yes
Legacy Posted Date
2018-11-16First Compliant Deposit (FCD) Date
2018-11-16Usage metrics
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