Microarray analysis during adipogenesis identifies new genes altered by antiretroviral drugs.
OBJECTIVE
To elucidate the pathogenesis of HAART-associated lipodystrophy, by investigating the effects of antiretroviral drugs on adipocyte differentiation and gene expression profile.


METHODS
Analysis of gene expression profile by DNA microarrays and quantitative RT-PCR of 3T3-L1 preadipocytes treated with the nucleoside reverse transcriptase inhibitors (NRTI) lamivudine, zidovudine, stavudine, and zalcitabine, and with the protease inhibitors (PI) indinavir, saquinavir, and lopinavir during maturation into adipocytes.


RESULTS
Under standard adipogenic differentiation protocols, PI significantly inhibited adipocyte differentiation, as demonstrated by cell viability assay and Oil Red O staining and quantification, whereas NRTI had mild effects on adipogenesis. Gene expression profile analysis showed that treatment with NRTI modulated the expression of transcription factors, such as Aebp1, Pou5f1 and Phf6, which could play a key role in the determination of the adipocyte phenotype. PI also modulated gene expression toward inhibition of adipocyte differentiation, with up-regulation of the Wnt signaling gene Wnt10a and down-regulation of the expression of genes encoding master adipogenic transcription factors (e.g., C/EBPalpha and PPARgamma), oestrogen receptor beta, and adipocyte-specific markers (e.g., Adiponectin, Leptin, Mrap, Cd36, S100A8).


CONCLUSIONS
This study identifies new genes modulated by PI and NRTI in differentiating adipocytes. Abnormal expression of these genes, which include master adipogenic transcription factors and genes involved in lipid metabolism and cell cycle control, could contribute to the understanding of the pathogenesis of HAART-associated lipodystrophy.
Pacenti
Monia
M
Department of Histology, Microbiology and Medical Biotechnologies, University of Padua, Via A. Gabelli 63, I-35121 Padua, Italy.
Barzon
Luisa
L
Favaretto
Francesca
F
Fincati
Karina
K
Romano
Sara
S
Milan
Gabriella
G
Vettor
Roberto
R
Palù
Giorgio
G
D016475:3T3 Cells; D017667:Adipocytes / Q000821:virology; D050156:Adipogenesis / Q000235:genetics*; D000818:Animals; D023241:Antiretroviral Therapy, Highly Active / Q000009:adverse effects*; D015415:Biomarkers; D002454:Cell Differentiation / Q000187:drug effects; D015536:Down-Regulation; D015870:Gene Expression; D017320:HIV Protease Inhibitors / Q000009:adverse effects; D039682:HIV-Associated Lipodystrophy Syndrome / Q000139:chemically induced*; D051379:Mice; D046228:Microarray Analysis / Q000379:methods; D018894:Reverse Transcriptase Inhibitors / Q000009:adverse effects; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods
D015415:Biomarkers; D017320:HIV Protease Inhibitors; D018894:Reverse Transcriptase Inhibitors