Targeting myofibroblasts in model systems of fibrosis by an artificial alpha-smooth muscle-actin promoter hybrid.
Myofibroblasts are the main cell types producing extracellular matrix proteins in a variety of fibrotic diseases. Therefore, they are useful targets for studies of intracellular communication and gene therapeutical approaches in scarring diseases. An artificial promoter containing the -702 bp regulatory sequence of the alpha-smooth muscle actin (SMA) gene linked to the first intron enhancer sequence of the beta-actin gene and the beta-globin intron-exon junction was constructed and tested for myofibroblast-dependent gene expression using the green fluorescent protein as a reporter. Reporter expression revealed myofibroblast-specific function in hepatic and renal myofibroblasts, in vitro. In addition, differentiation-dependent activation of the SMA-beta-actin promoter hybrid was shown after induction of myofibroblastic features in mesangial cells by stretching treatment. Furthermore, wound healing experiments with SMA-beta-actin promoter reporter mice demonstrated myofibroblast-specific action, in vivo. In conclusion, the -702 bp regulatory region of the SMA promoter linked to enhancing beta-actin and beta-globin sequences benefits from its small size and is suggested as a promising tool to target myofibroblasts as the crucial cell type in various scarring processes.
Hirschfeld
Julia
J
Institute of Pathology, University Hospital Cologne, Koeln, Germany.
Maurer
Julia
J
Jung
Diana
D
Kwiecinski
Monika
M
Khimji
Al Karim
AK
Dienes
Hans Peter
HP
Fries
Jochen W U
JW
Odenthal
Margarete
M
D000199:Actins / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D004195:Disease Models, Animal; D005355:Fibrosis / Q000235:genetics* / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods*; D051379:Mice; D008822:Mice, Transgenic; D032446:Myoblasts / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D015202:Protein Engineering / Q000379:methods; D014162:Transfection / Q000379:methods
D000199:Actins; C544013:alpha-smooth muscle actin, mouse