Generation of expression constructs that secrete bioactive αMSH and their use in the treatment of experimental autoimmune encephalomyelitis

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Abstract

α Melanocyte-stimulating hormone (αMSH) is a 13 amino acid peptide with potent anti-inflammatory effects. We created two DNA expression constructs (miniPOMC and pACTH1–17) that encode bioactive versions of the αMSH peptide, and tested these constructs for therapeutic effects in experimental autoimmune encephalomyelitis (EAE). Each construct contained the sequences for αMSH, as well as the sequences that are involved in the secretion and processing of the POMC gene with the assumption that these sequences would promote processing and release of the encoded αMSH peptide. The differences between the two constructs lie at the C-terminal end where amino acids necessary for amidation of αMSH were included in only the pACTH1–17 construct. These two constructs were tested in vitro in bioassays, and in vivo in a mouse model of EAE. The results show that although bioactive peptides are secreted from cells transfected with either construct, there appears to be a significant therapeutic effect only with the pACTH1–17 construct which contains the extra C-terminal amino acids. The data suggest that it is possible to engineer DNA expression vectors encoding small secreted peptides such as αMSH, and that similar type constructs may be useful as therapeutics for the treatment of inflammatory diseases.

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Acknowledgements

We thank Ms. Rene Hallack for her work in purifying DNA constructs. We also thank Dr Andy Tomlinson and Dr Jennie Lill for their efforts in HPLC and mass spectrometry analyses and Dr Janet Lathey for offering advice concerning preparation of the manuscript.

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Correspondence to M L Hedley.

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Keywords

  • αMSH
  • inflammation
  • gene therapy
  • NF-κB
  • EAE

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