cDNA microarray analysis reveals new candidate genes possibly linked to side effects under mycophenolate mofetil therapy.
BACKGROUND
Mycophenolate mofetil (MMF) the prodrug of mycophenolic acid is usually well tolerated. Side effects such as anemia and diarrhea occur in approximately 10%-15% of patients. The aim of this study was to examine in a rat model the effect of MMF on gene expression in liver and gut to identify target genes with possible relevance to MMF side effects.


METHODS
Twelve Wistar rats were treated with 40 mg/kg body weight MMF orally for 21 days. Controls (n=9) received vehicle only. RNA was extracted from liver, jejunum, ileum, and colon and transcribed into cDNA. Regulated genes were identified in liver by DNA microarray experiments. Gene regulation was verified in liver and gut using quantitative real-time PCR on the LightCycler instrument. Transcription elongation factor 2 served as reference gene.


RESULTS
Microarray analysis revealed that major alpha-hemoglobin, polymeric immunoglobulin receptor, catalase, and CCAAT/enhancer protein alpha gene expression were down-regulated in livers of MMF-treated rats 10-, 5.5-, 4-, and 5-fold, respectively. These findings could be confirmed through quantitative real-time PCR analysis of gene expression in liver, ileum, jejunum, and colon.


CONCLUSIONS
Using microarray analysis and a rat model four candidate genes which may be functionally linked to side effects (major alpha-hemoglobin-->anaemia; polymeric immunoglobulin receptor-->protection of mucosa; catalase and CCAAT/enhancer protein alpha-->oxidative stress) of MMF therapy were identified.
Shipkova
Maria
M
Zentralinstitut für Klinische Chemie und Laboratoriumsmedizin, Klinikum Stuttgart, Katharinenhospital, Kriegsbergstrasse 60, D-70174 Stuttgart, Germany. m.shipkova@klinikum-stuttgart.de
Spielbauer
Bettina
B
Voland
Antje
A
Gröne
Hermann-Josef
HJ
Armstrong
Victor W
VW
Oellerich
Michael
M
Wieland
Eberhard
E
D000818:Animals; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics; D002374:Catalase / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects*; D020869:Gene Expression Profiling; D006454:Hemoglobins / Q000235:genetics; D007166:Immunosuppressive Agents / Q000009:adverse effects*; D007413:Intestinal Mucosa / Q000378:metabolism; D008099:Liver / Q000378:metabolism; D009173:Mycophenolic Acid / Q000009:adverse effects* / Q000031:analogs & derivatives*; D020411:Oligonucleotide Array Sequence Analysis; D020033:Protein Isoforms / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D019056:Receptors, Polymeric Immunoglobulin / Q000235:genetics
D022762:CCAAT-Enhancer-Binding Proteins; D006454:Hemoglobins; D007166:Immunosuppressive Agents; D020033:Protein Isoforms; D019056:Receptors, Polymeric Immunoglobulin; D002374:Catalase; D009173:Mycophenolic Acid