Generation of expression constructs that secrete bioactive alphaMSH and their use in the treatment of experimental autoimmune encephalomyelitis.
alpha Melanocyte-stimulating hormone (alphaMSH) 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 alphaMSH peptide, and tested these constructs for therapeutic effects in experimental autoimmune encephalomyelitis (EAE). Each construct contained the sequences for alphaMSH, 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 alphaMSH peptide. The differences between the two constructs lie at the C-terminal end where amino acids necessary for amidation of alphaMSH 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 alphaMSH, and that similar type constructs may be useful as therapeutics for the treatment of inflammatory diseases.
Yin
P
P
Zycos Inc, Lexington, MA 02421, USA.
Luby
T M
TM
Chen
H
H
Etemad-Moghadam
B
B
Lee
D
D
Aziz
N
N
Ramstedt
U
U
Hedley
M L
ML
D000324:Adrenocorticotropic Hormone / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000628:therapy*; D005260:Female; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D008546:Melanoma, Experimental; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D011333:Pro-Opiomelanocortin / Q000235:genetics; D014162:Transfection / Q000379:methods*; D000521:alpha-MSH / Q000235:genetics* / Q000378:metabolism
D016328:NF-kappa B; D000521:alpha-MSH; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone