Matrix Metalloproteinase Responsive Delivery of Myostatin Inhibitors
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The inhibition of myostatin - a member of the transforming growth factor (TGF–β) family - drives regeneration of functional skeletal muscle tissue. We developed a bioresponsive drug delivery system (DDS) linking release of a myostatin inhibitor (MI) to inflammatory flares of myositis to provide self-regulated MI concentration gradients within tissues of need.
A protease cleavable linker (PCL) – responding to MMP upregulation – is attached to the MI and site-specifically immobilized on microparticle surfaces.
The PCL disintegrated in a matrix metalloproteinase (MMP) 1, 8, and particularly MMP-9 concentration dependent manner, with MMP-9 being an effective surrogate biomarker correlating with the activity of myositis. The bioactivity of particle-surface bound as well as released MI was confirmed by luciferase suppression in stably transfected HEK293 cells responding to myostatin induced SMAD phosphorylation.
We developed a MMP-responsive DDS for MI delivery responding to inflammatory flare of a diseased muscle matching the kinetics of MMP-9 upregulation, with MMP-9 kinetics matching (patho-) physiological myostatin levels.
KEY WORDSbioorthogonal chemistry bioresponsive drug delivery controlled release myostatin protease cleavable linker
Activin receptor IIB
Bovine serum albumin
Copper(I)-catalyzed azide-alkyne cycloaddition
Drug delivery system
Duchenne muscular dystrophy
Growth differentiation factor 8
High-performance liquid chromatography
Insulin-like growth factor I
Matrix-assisted laser desorption ionization mass spectrometry
Myosin heavy chain
Nuclear factor-kappa B
Protease cleavable linker
Relative light unit
Real time polymerase chain reaction
SMAD binding element
Strain-promoted azide-alkyne cycloaddition
Solid phase peptide synthesis
Transglutaminase (human, fXIIIa)
Transforming growth factor beta
Tumor necrosis factor α
ACKNOWLEDGMENTS AND DISCLOSURES
We thank Petra Knaus (Freie Universität Berlin, Germany) for providing us with the pGl3ti-SBE constructs and Melanie Krug for excellent technical assistance. We thank Dr. Joachim Nickel for providing C2C12 myoblast cells. The financial support of the Bavarian research foundation (grant # AZ-1044-12 ‘FORMOsA’) and the Deutsche Forschungsgemeinschaft (DFG; ME 3820/3-1) are gratefully acknowledged. H.G. is full time associate of Gilyos GmbH.
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