Effects of mycophenolic acid on human fibroblast proliferation, migration and adhesion in vitro and in vivo.
Mycophenolic acid (MPA) is a potent inhibitor of the inosine monophosphate dehydrogenase and used as an immunosuppressive drug in transplantation. MPA inhibits proliferation of T- and B-lymphocytes by guanosine depletion. Since fibroblasts rely on the de novo synthesis of guanosine nucleotides, it is assumed that MPA interacts with fibroblasts causing an increased frequency of wound healing problems. We show a downregulation of the cytoskeletal proteins vinculin, actin and tubulin in fibroblasts exposed to pharmacological doses of MPA using microarray technology, real-time polymerase chain reaction (PCR) and Western blot. This reduction in RNA and protein content is accompanied by a substantial rearrangement of the cytoskeleton in MPA-treated fibroblasts as documented by immunofluorescence. The dysfunctional fibroblast growth was validated by scratch test documenting impaired migrational capacity. In contrast, cell adhesion was increased in MPA-treated fibroblasts. The results of the cultured human fibroblasts were applied to skin biopsies of renal transplant recipients. Skin biopsies of patients treated with MPA expressed less vinculin, actin and tubulin as compared to control biopsies that could explain potential wound healing problems posttransplantation. The perspective of MPA-induced cytoskeletal dysfunction may go beyond wound healing disturbances and may have beneficial effects on (renal) allografts with respect to scarring.
Morath
C
C
Department of Nephrology, University of Heidelberg, Germany. christian.morath@med.uni-heidelberg.de
Reuter
H
H
Simon
V
V
Krautkramer
E
E
Muranyi
W
W
Schwenger
V
V
Goulimari
P
P
Grosse
R
R
Hahn
M
M
Lichter
P
P
Zeier
M
M
D001706:Biopsy; D002232:Carbocyanines / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D062109:Dermatologic Surgical Procedures; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects*; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007211:Indoles / Q000378:metabolism; D009173:Mycophenolic Acid / Q000494:pharmacology*; D010590:Phalloidine / Q000378:metabolism; D012235:Rhodamines / Q000378:metabolism; D012867:Skin / Q000166:cytology
D002232:Carbocyanines; D005456:Fluorescent Dyes; D007166:Immunosuppressive Agents; D007211:Indoles; D012235:Rhodamines; C089187:cyanine dye 3; D010590:Phalloidine; C009434:tetramethylrhodamine isothiocyanate; C007293:DAPI; D009173:Mycophenolic Acid; D016650:Fluorescein-5-isothiocyanate