Latency can be conferred to a variety of cytokines by fusion with latency-associated peptide from TGF-beta

TY - JOUR

T1 - Latency can be conferred to a variety of cytokines by fusion with latency-associated peptide from TGF-beta

AU - Mullen, Lisa

AU - Rigby, Anne

AU - Sclanders, Michelle

AU - Adams, Gill

AU - Mittal, Gayatri

AU - Colston, Julia

AU - Fatah, Rewas

AU - Subang, Cristina

AU - Foster, Julie

AU - Francis-West, Philippa

AU - Koester, Mario

AU - Hauser, Hansjoerg

AU - Layward, Lorna

AU - Vessillier, Sandrine

AU - Annenkov, Alex

AU - Al-Izki, Sarah

AU - Pryce, Gareth

AU - Bolton, Chris

AU - Baker, David

AU - Gould, David J.

AU - Chernajovsky, Yuti

PY - 2014/1

Y1 - 2014/1

N2 - Objectives: Targeting cytokines to sites of disease has clear advantages because it increases their therapeutic index. We designed fusion proteins of the latent-associated peptide (LAP) derived from TGF-beta with various cytokines via a matrix metalloproteinase (MMP) cleavage site. This design confers latency, increased half-life and targeting to sites of inflammation. The aim of this study is to determine whether this approach can be applied to cytokines of different molecular structures and sizes.Methods: Mature cytokines cloned downstream of LAP and a MMP cleavage site were expressed in 293T cells and assessed for latency and biological activity by Western blotting and bioassay.Results: We demonstrate here that fusion proteins of TGF-beta, erythropoietin, IL-1ra, IL-10, IL-4, BMP-7, IGF1 and IL-17 were rendered latent by fusion to LAP, requiring cleavage to become active in respective bioassays. As further proof of principle, we also show that delivery of engineered TGF-beta can inhibit experimental autoimmune encephalomyelitis and that this approach can be used to efficiently deliver cytokines to the brain and spinal cord in mice with this disease.Conclusions: The latent cytokine approach can be successfully applied to a range of molecules, including cytokines of different molecular structure and mass, growth factors and a cytokine antagonist.

AB - Objectives: Targeting cytokines to sites of disease has clear advantages because it increases their therapeutic index. We designed fusion proteins of the latent-associated peptide (LAP) derived from TGF-beta with various cytokines via a matrix metalloproteinase (MMP) cleavage site. This design confers latency, increased half-life and targeting to sites of inflammation. The aim of this study is to determine whether this approach can be applied to cytokines of different molecular structures and sizes.Methods: Mature cytokines cloned downstream of LAP and a MMP cleavage site were expressed in 293T cells and assessed for latency and biological activity by Western blotting and bioassay.Results: We demonstrate here that fusion proteins of TGF-beta, erythropoietin, IL-1ra, IL-10, IL-4, BMP-7, IGF1 and IL-17 were rendered latent by fusion to LAP, requiring cleavage to become active in respective bioassays. As further proof of principle, we also show that delivery of engineered TGF-beta can inhibit experimental autoimmune encephalomyelitis and that this approach can be used to efficiently deliver cytokines to the brain and spinal cord in mice with this disease.Conclusions: The latent cytokine approach can be successfully applied to a range of molecules, including cytokines of different molecular structure and mass, growth factors and a cytokine antagonist.

KW - bioassay

KW - cytokines

KW - latency-associated peptide

KW - protein design

KW - targeted delivery

KW - GROWTH-FACTOR-BETA

KW - NF-KAPPA-B

KW - BINDING

KW - SITES

KW - ACTIVATION

KW - EXPRESSION

KW - BIOASSAY

U2 - 10.1517/17425247.2013.839655

DO - 10.1517/17425247.2013.839655

M3 - Article

SN - 1742-5247

VL - 11

SP - 5

EP - 16

JO - Expert Opinion On Drug Delivery

JF - Expert Opinion On Drug Delivery

IS - 1

ER -