Maternal obesity during pregnancy leads to adipose tissue ER stress in mice via miR-126-mediated reduction in Lunapark

Autor: Asha A. M. Carpenter, Lucas Carminatti Pantaleão, Thomas Prates Ong, Robin Antrobus, Elena Loche, Laura C. Kusinski, Susan E. Ozanne, Denise S. Fernandez-Twinn, Thomas J. Ashmore, Daniella Duque-Guimarães, Juliana de Almeida-Faria, Martin Bushell
Rok vydání: 2021
Předmět:
Blood Glucose
0301 basic medicine
Endocrinology
Diabetes and Metabolism
Adipose tissue
Obesity
Maternal
Mice
chemistry.chemical_compound
0302 clinical medicine
Pregnancy
Adipocyte
Adipocytes
EIF2AK3
Glucose metabolism
Gene knockdown
Endoplasmic Reticulum Stress
Phenotype
Adipose Tissue
Prenatal Exposure Delayed Effects
Female
ER stress
Signal Transduction
medicine.medical_specialty
XBP1
Offspring
Down-Regulation
BIOINFORMÁTICA
030209 endocrinology & metabolism
Biology
Lunapark
Article
Nutritional programming
03 medical and health sciences
Insulin resistance
Maternal obesity
3T3-L1 Cells
Internal medicine
miR-126-3p
Internal Medicine
medicine
Animals
Homeodomain Proteins
medicine.disease
Mice
Inbred C57BL
Disease Models
Animal
MicroRNAs
030104 developmental biology
Endocrinology
chemistry
Insulin Receptor Substrate Proteins
Unfolded protein response
Insulin Resistance
Zdroj: Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual)
Universidade de São Paulo (USP)
instacron:USP
Diabetologia
ISSN: 1432-0428
0012-186X
DOI: 10.1007/s00125-020-05357-4
Popis: 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. Graphical abstract
Databáze: OpenAIRE
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