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Molecular Medicine

, Volume 21, Issue 1, pp 442–452 | Cite as

d-Amino Acid Substitution of Peptide-Mediated NF-κB Suppression in mdx Mice Preserves Therapeutic Benefit in Skeletal Muscle, but Causes Kidney Toxicity

  • Daniel P. Reay
  • Sheldon I. Bastacky
  • Kathryn E. Wack
  • Donna B. Stolz
  • Paul D. Robbins
  • Paula R. Clemens
Research Article

Abstract

In Duchenne muscular dystrophy (DMD) patients and the mdx mouse model of DMD, chronic activation of the classical nuclear factor-κB (NF-κB) pathway contributes to the pathogenesis that causes degeneration of muscle fibers, inflammation and fibrosis. Prior studies demonstrate that inhibition of inhibitor of κB kinase (IKK)-mediated NF-κB activation using l-isomer NF-κB essential modulator (NEMO)-binding domain (NBD) peptide-based approaches reduce muscle pathology in the mdx mouse. For our studies, the NBD peptide is synthesized as a fusion peptide with an eight-lysine (8K) protein transduction domain to facilitate intracellular delivery. We hypothesized that the d-isoform peptide could have a greater effect than the naturally occurring l-isoform peptide due to the longer persistence of the d-isoform peptide in vivo. In this study, we compared systemic treatment with low (1 mg/kg) and high (10 mg/kg) doses of l- and d-isomer 8K-wild-type-NBD peptide in mdx mice. Treatment with both l- or d-isoform 8K-wild-type-NBD peptide resulted in decreased activation of NF-κB and improved histology in skeletal muscle of the mdx mouse. However, we observed kidney toxicity (characterized by proteinuria), increased serum creatinine, activation of NF-κB and pathological changes in kidney cortex that were most severe with treatment with the d-isoform of 8K-wild-type-NBD peptide. The observed toxicity was also seen in normal mice.

Notes

Acknowledgments

This work was supported in part by by the Kidney Imaging Core of the Pittsburgh Center for Kidney Research (NIH grant P30-DK079307). In addition, some of the transmission electron microscopy equipment used in this study was funded by grant # 1S10RR019003-01. The work was funded by the Department of Veterans Affairs (VA) Medical Center Merit Review Grant and departmental funds (PRC). We would like to thank Katie Clark and the UPMC Presbyterian Hospital Pathology Department Electron Microscopy Laboratory for their contributions to this manuscript.

Supplementary material

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Authors and Affiliations

  • Daniel P. Reay
    • 1
    • 2
  • Sheldon I. Bastacky
    • 3
  • Kathryn E. Wack
    • 4
  • Donna B. Stolz
    • 4
    • 5
  • Paul D. Robbins
    • 6
  • Paula R. Clemens
    • 1
    • 2
  1. 1.Department of Veterans Affairs Medical CenterNeurology ServicePittsburghUSA
  2. 2.S520 Biomedical Science Tower, Department of NeurologyUniversity of PittsburghPittsburghUSA
  3. 3.Department of PathologyUniversity of PittsburghPittsburghUSA
  4. 4.Department of Cell BiologyUniversity of PittsburghPittsburghUSA
  5. 5.Center for Biologic ImagingUniversity of PittsburghPittsburghUSA
  6. 6.Department of Metabolism and AgingScripps FloridaJupiterUSA

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