The small molecule phenamil is a modulator of adipocyte differentiation and PPARgamma expression.
We previously described the use of a cell-based screening approach to identify small molecules that regulate adipocyte differentiation. Here we identify the amiloride derivative phenamil as an adipogenic compound. Phenamil acutely induces expression of the key transcription factor of adipogenesis, peroxisome proliferator-activated receptor gamma (PPARgamma) and, consequently, promotes the differentiation of multiple preadipocyte cell lines, including 3T3-L1 and F442A. Interestingly, the adipogenic action of phenamil is distinct from and additive with both PPARgamma ligands and the previously identified adipogenic small molecule harmine. To identify signaling pathways mediating phenamil's effects, we performed transcriptional profiling of 3T3-F442A preadipocytes. ETS variant 4 (ETV4) was identified as a gene rapidly induced by phenamil but not by other adipogenic small molecules or PPARgamma agonists. Transient expression of ETV4 in preadipocytes enhances the expression of PPARgamma. Stable overexpression of ETV4 promotes expression of PPARgamma and its downstream target genes and enhances morphological differentiation. Finally, knockdown of PPARgamma expression by shRNA blocks the effects of phenamil on adipocyte differentiation and gene expression, but it does not block phenamil induction of ETV4, which suggests that ETV4 acts upstream of PPARgamma in differentiation processes. These results identify a phenamil as new small molecule tool for the probing of adipocyte differentiation that acts, at least in part, through induction of ETV4 expression.
Howard Hughes Medical Institute, University of California at Los Angeles, Los Angeles, CA, USA.
Kye Won
KW
Park
Hironori
H
Waki
Sung-Pil
SP
Choi
Ki-Moon
KM
Park
Peter
P
Tontonoz
D016475:3T3 Cells; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D000584:Amiloride / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D000818:Animals; D051743:CCAAT-Enhancer-Binding Protein-delta / Q000737:chemistry / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006247:Harmine / Q000378:metabolism; D051379:Mice; D015394:Molecular Structure; D008996:Monoamine Oxidase Inhibitors / Q000378:metabolism; D010080:Oxazoles / Q000737:chemistry / Q000494:pharmacology; D047495:PPAR gamma / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D050783:Proto-Oncogene Proteins c-ets / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D026941:Sodium Channel Blockers / Q000737:chemistry / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology
D004268:DNA-Binding Proteins; C540571:Etv4 protein, mouse; C513242:Etv5 protein, mouse; D008996:Monoamine Oxidase Inhibitors; D010080:Oxazoles; D047495:PPAR gamma; D050783:Proto-Oncogene Proteins c-ets; D026941:Sodium Channel Blockers; D014157:Transcription Factors; C013407:benzamil; D051743:CCAAT-Enhancer-Binding Protein-delta; C047088:phenylamil; C401311:GW 7845; D014443:Tyrosine; D006247:Harmine; D000584:Amiloride