Mullen, Lisa, Adams, Gill, Foster, Julie, Vessillier, Sandrine, Köster, Mario, Hauser, Hansjörg, Layward, Lorna, Gould, David and Chernajovsky, Yuti (2014) A comparative study of matrix metalloproteinase and aggrecanase mediated release of latent cytokines at arthritic joints. Annals of the Rheumatic Diseases, 73 (9). pp. 1728-1736. ISSN 0003-4967

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Abstract

BACKGROUND:

Latent cytokines are engineered by fusing the latency associated peptide (LAP) derived from transforming growth factor-β (TGF-β) with the therapeutic cytokine, in this case interferon-β (IFN-β), via an inflammation-specific matrix metalloproteinase (MMP) cleavage site.

OBJECTIVES:

To demonstrate latency and specific delivery in vivo and to compare therapeutic efficacy of aggrecanase-mediated release of latent IFN-β in arthritic joints to the original MMP-specific release.

METHODS:

Recombinant fusion proteins with MMP, aggrecanase or devoid of cleavage site were expressed in CHO cells, purified and characterised in vitro by Western blotting and anti-viral protection assays. Therapeutic efficacy and half-life were assessed in vivo using the mouse collagen-induced arthritis model (CIA) of rheumatoid arthritis and a model of acute paw inflammation, respectively. Transgenic mice with an IFN-regulated luciferase gene were used to assess latency in vivo and targeted delivery to sites of disease.

RESULTS:

Efficient localised delivery of IFN-β to inflamed paws, with low levels of systemic delivery, was demonstrated in transgenic mice using latent IFN-β. Engineering of latent IFN-β with an aggrecanase-sensitive cleavage site resulted in efficient cleavage by ADAMTS-4, ADAMTS-5 and synovial fluid from arthritic patients, with an extended half-life similar to the MMP-specific molecule and greater therapeutic efficacy in the CIA model.

CONCLUSIONS:

Latent cytokines require cleavage in vivo for therapeutic efficacy, and they are delivered in a dose dependent fashion only to arthritic joints. The aggrecanase-specific cleavage site is a viable alternative to the MMP cleavage site for the targeting of latent cytokines to arthritic joints.

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 14:34
Last Modified:	16 Nov 2018 14:35
URI:	http://srodev.sussex.ac.uk/id/eprint/80266

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