Mirra, Paola (2008) The PED/PEA-15 diabetogene as a potential thiazolidinedione target in type 2 diabetes treatment. [Tesi di dottorato] (Unpublished)

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Item Type:	Tesi di dottorato
Resource language:	English
Title:	The PED/PEA-15 diabetogene as a potential thiazolidinedione target in type 2 diabetes treatment
Creators:	Creators Email Mirra, Paola paolamirra.lib@libero.it
Date:	9 December 2008
Number of Pages:	93
Institution:	Università degli Studi di Napoli Federico II
Department:	Biologia e patologia cellullare e molecolare "L. Califano"
Scuola di dottorato:	Medicina molecolare
Dottorato:	Oncologia ed endocrinologia molecolare
Ciclo di dottorato:	21
Coordinatore del Corso di dottorato:	nome email Vecchio, Giancarlo vecchio@unina.it
Tutor:	nome email Beguinot, Francesco beguino@unina.it
Date:	9 December 2008
Number of Pages:	93
Keywords:	Type 2 diabetes; Thiazolidinediones; ped/pea-15
Settori scientifico-disciplinari del MIUR:	Area 06 - Scienze mediche > MED/04 - Patologia generale
Date Deposited:	17 Nov 2009 09:49
Last Modified:	30 Apr 2014 19:36
URI:	http://www.fedoa.unina.it/id/eprint/3408

Collection description

The phosphoprotein enriched in diabetes/phosphoprotein enriched in astrocytes (PED/PEA-15) gene is over-expressed in tissues from individuals with type 2 diabetes. In cultured cells and in vivo, ped/pea-15 over-expression impairs insulin activation of protein kinase C zeta (PKCζ) and glucose disposal. Indeed, transgenic mice ubiquitously over-expressing ped/pea-15 feature impaired glucose tolerance and insulin resistance. However, it is still unknown how PED/PEA-15 gene expression is regulated. Recently, we have obtained evidence that Thiazolidinediones (TZDs), a class of peroxisome proliferator-activated receptor gamma (PPARγ) ligands, repress ped/pea-15 expression both at mRNA and protein levels. PPARγ is a member of the nuclear hormone receptor super-family that modulates gene expression upon ligand binding. Ligand-mediated activation of PPARγ has been linked to glucose homeostasis, cellular differentiation, apoptosis and anti-inflammatory responses. The aim of this work was to study whether and how TZD-activated PPARγ may exert its action on ped/pea-15 expression. In L6 skeletal muscle cells, rosiglitazone (RGTZ) decreased both ped/pea-15 mRNA and protein levels. This was paralleled by the RGTZ-dependent reduction of PKCα activation and a consequent increase of PKCζ activity. Consistent with the in vitro data, muscle tissue from control mice showed a 40% decrease of ped/pea-15 protein levels upon 10 days of treatment with RGTZ. HeLa cells treated with TPA, the prototypical AP1 activator, showed an increase in PED/PEA-15 mRNA levels. TPA induced also the activation of PED/PEA-15 promoter activity measured by luciferase assays. Interestingly, TPA effect was suppressed by RGTZ. EMSA and ChIP assays revealed that TPA caused an increased binding to the CRE-like site (a putative AP1 binding site) on the PED/PEA-15 promoter, and this binding was reduced by RGTZ. Since PPARγ activation by TZDs blocks AP1-mediated gene transcription, we hypothesized that PPARγ transrepression of the PED/PEA-15 gene could be due to a competition for limiting amounts of co-activators present in the cell. Indeed, the suppressive effect of rosiglitazone on PED/PEA-15 gene expression was blocked by over-expressing a dominant negative form of PPARγ, which lacks the ability to recruit the coactivator CBP/p300 (CREB-binding protein). The over-expression of p300 did not rescue the cells from the suppressive effect of PPARγ on PED/PEA-15 promoter activity, suggesting that PPARγ-mediated repression of AP1 transcriptional activity occurs in a CBP/p300-independent manner. Taken together, these results indicate that regulation through the CRE-like site on PED/PEA-15 promoter induces PED/PEA-15 transcription. This effect was blocked by treatment with TZDs. Finally, the results of this study identified PED/PEA-15 gene as a potential target for TZDs therapeutic action.

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