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de Almeida-Faria, Juliana ^{[1, 2, 3]} ; Duque-Guimaraes, Daniella E. ^{[1, 2]} ; Ong, Thomas P. ^{[4, 1, 2]} ; Pantaleao, Lucas C. ^{[1, 2]} ; Carpenter, Asha A. ^{[1, 2]} ; Loche, Elena ^{[1, 2]} ; Kusinski, Laura C. ^{[1, 2]} ; Ashmore, Thomas J. ^{[1, 2]} ; Antrobus, Robin ^[5] ; Bushell, Martin ^[6] ; Fernandez-Twinn, Denise S. ^{[1, 2]} ; Ozanne, Susan E. ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Univ Cambridge, Metab Res Labs, Cambridge - England

^[2] Addenbrookes Hosp, Wellcome Trust MRC Inst Metab Sci, MRC Metab Dis Unit, Cambridge - England

^[3] Univ Estadual Campinas, Fac Med Sci, Obes & Comorbid Res Ctr, Sao Paulo - Brazil

^[4] Univ Sao Paulo, Food Res Ctr, Fac Pharmaceut Sci, Dept Food & Expt Nutr, Sao Paulo - Brazil

^[5] Univ Cambridge, Cambridge Inst Med Res, Hills Rd, Cambridge - England

^[6] Beatson Inst, Canc Res UK CRUK, Glasgow, Lanark - Scotland

• Tipo de documento: Artigo Científico
• Fonte: Diabetologia; v. 64, n. 4 JAN 2021.
• Citações Web of Science: 0

Resumo

Aims/hypothesis Levels of the microRNA (miRNA) miR-126-3p are programmed cell-autonomously in visceral adipose tissue of adult offspring born to obese female C57BL/6J mice. The spectrum of miR-126-3p targets and thus the consequences of its dysregulation for adipocyte metabolism are unknown. Therefore, the aim of the current study was to identify novel targets of miR-126-3p in vitro and then establish the outcomes of their dysregulation on adipocyte metabolism in vivo using a well-established maternal obesity mouse model. Methods miR-126-3p overexpression in 3T3-L1 pre-adipocytes followed by pulsed stable isotope labelling by amino acids in culture (pSILAC) was performed to identify novel targets of the miRNA. Well-established bioinformatics algorithms and luciferase assays were then employed to confirm those that were direct targets of miR-126-3p. Selected knockdown experiments were performed in vitro to define the consequences of target dysregulation. Quantitative real-time PCR, immunoblotting, histology, euglycaemic-hyperinsulinaemic clamps and glucose tolerance tests were performed to determine the phenotypic and functional outcomes of maternal programmed miR-126-3p levels in offspring adipose tissue. Results The proteomic approach confirmed the identity of known targets of miR-126-3p (including IRS-1) and identified Lunapark, an endoplasmic reticulum (ER) protein, as a novel one. We confirmed by luciferase assay that Lunapark was a direct target of miR-126-3p. Overexpression of miR-126-3p in vitro led to a reduction in Lunapark protein levels and increased Perk (also known as Eif2ak3) mRNA levels and small interference-RNA mediated knockdown of Lunapark led to increased Xbp1, spliced Xbp1, Chop (also known as Ddit3) and Perk mRNA levels and an ER stress transcriptional response in 3T3-L1 pre-adipocytes. Consistent with the results found in vitro, increased miR-126-3p expression in adipose tissue from adult mouse offspring born to obese dams was accompanied by decreased Lunapark and IRS-1 protein levels and increased markers of ER stress. At the whole-body level the animals displayed glucose intolerance. Conclusions/interpretation Concurrently targeting IRS-1 and Lunapark, a nutritionally programmed increase in miR-126-3p causes adipose tissue insulin resistance and an ER stress response, both of which may contribute to impaired glucose tolerance. These findings provide a novel mechanism by which obesity during pregnancy leads to increased risk of type 2 diabetes in the offspring and therefore identify miR-126-3p as a potential therapeutic target. (AU)

• Processo FAPESP: 14/17012-4 - A Pharmacological intervention to prevent the effects of maternal obesity on offspring adipose tissue insulin resistance
• Beneficiário: Juliana de Almeida Faria
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
• Processo FAPESP: 17/03525-8 - O papel da obesidade materna na modulação dos microRNAs hipotalâmicos da prole e consequências metabólicas associadas
• Beneficiário: Juliana de Almeida Faria
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 14/20380-5 - Envolvimento dos microRNAs no processo de envelhecimento precoce causado pela obesidade
• Beneficiário: Daniella Esteves Duque Guimarães
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 13/07914-8 - FoRC - Centro de Pesquisa em Alimentos
• Beneficiário: Bernadette Dora Gombossy de Melo Franco
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs