title: Overexpression of repressive cAMP response element modulators in high glucose and fatty acid-treated rat islets. A common mechanism for glucose toxicity and lipotoxicity?
abstract: The hyperlipidemia and hyperglycemia of the diabetic state accelerate beta-cell dysfunction, yet the mechanisms are not fully defined. We used rat islet-specific oligonucleotide arrays (Metabolex Rat Islet Genechips) to identify genes that are coordinately regulated by high glucose and free fatty acids (FFA). Exposure of rat islets to FFA (125 microM for 2 days) or glucose (27 mM for 4 days) reduced glucose-stimulated insulin secretion by 70 +/- 5 and 40 +/- 4%, respectively, relative to control-cultured islets. These treatments also substantially reduced the insulin content of the islets. Islet Genechips analysis revealed that the mRNA levels of cAMP response element modulator (CREM)-17X and inducible cAMP early repressor were significantly increased in both 27 mM glucose- and FFA-treated islets. Removing FFA or high glucose from the culture medium restored glucose-stimulated insulin secretion and the mRNA levels of the two CREM repressors to normal. Northern blot analysis revealed a 5-fold increase in the abundance of CREM-17X mRNA and a concomitant 50% reduction in the insulin mRNA in FFA-treated islets. Transient transfection of the insulin-secreting beta HC9 cells with CREM-17X suppressed rat insulin promoter activity by nearly 50%. Overexpression of CREM-17X in intact islets via adenovirus infection decreased islet insulin mRNA levels and insulin content and resulted in a significant decrease in glucose- or KCl-induced insulin secretion. Taken together, these data suggest that up-regulation of CREM repressors by either FFA or high glucose exacerbates beta-cell failure in type 2 diabetes by suppressing insulin gene transcription.
authors.affiliation: Department of Insulin Secretion Genomics, Metabolex, Inc., Hayward, California 94545, USA. yunping_zhou@merck.com
authors.forename: Yun-Ping
authors.initials: YP
authors.lastname: Zhou
authors.forename: Kimberly
authors.initials: K
authors.lastname: Marlen
authors.forename: John F
authors.initials: JF
authors.lastname: Palma
authors.forename: Anthony
authors.initials: A
authors.lastname: Schweitzer
authors.forename: Linda
authors.initials: L
authors.lastname: Reilly
authors.forename: Francine M
authors.initials: FM
authors.lastname: Gregoire
authors.forename: Gann G
authors.initials: GG
authors.lastname: Xu
authors.forename: John E
authors.initials: JE
authors.lastname: Blume
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Johnson
mesh_terms: D000818:Animals; D051786:Cyclic AMP Response Element Modulator; D004268:DNA-Binding Proteins / Q000096:biosynthesis*; D005227:Fatty Acids / Q000494:pharmacology*; D020869:Gene Expression Profiling; D005947:Glucose / Q000494:pharmacology*; D066298:In Vitro Techniques; D007328:Insulin / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000096:biosynthesis; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics; D015533:Transcriptional Activation / Q000187:drug effects*
chemical_list: D004268:DNA-Binding Proteins; D005227:Fatty Acids; D007328:Insulin; D012333:RNA, Messenger; D012097:Repressor Proteins; D014157:Transcription Factors; D051786:Cyclic AMP Response Element Modulator; D005947:Glucose