Maternal obesity during pregnancy leads to adipose tissue ER stress in mice via miR-126-mediated reduction in Lunapark.
Maternal obesity during pregnancy leads to adipose tissue ER stress in mice via miR-126-mediated reduction in Lunapark.
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DOI：
10.1007/s00125-020-05357-4
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发表时间：
2021-04
期刊：
影响因子：
8.2
通讯作者：
Ozanne SE
中科院分区：
文献类型：
--
作者：
de Almeida-Faria J;Duque-Guimarães DE;Ong TP;Pantaleão LC;Carpenter AA;Loche E;Kusinski LC;Ashmore TJ;Antrobus R;Bushell M;Fernandez-Twinn DS;Ozanne SE
microRNA (miRNA) miR-126-3p水平在肥胖雌性C57BL/6J小鼠成年后代的内脏脂肪组织中是细胞自主编程的。miR-126-3p靶点的谱及其对脂肪细胞代谢失调的后果尚不清楚。因此，本研究的目的是在体外鉴定miR-126-3p的新靶点，然后通过建立成熟的母体肥胖小鼠模型，建立其体内脂肪细胞代谢失调的结果。miR-126-3p在3T3-L1前脂肪细胞中过表达，然后通过培养氨基酸（pSILAC）进行脉冲稳定同位素标记，以鉴定miRNA的新靶标。然后采用成熟的生物信息学算法和荧光素酶测定来确认miR-126-3p的直接靶点。在体外进行了选择性敲低实验，以确定目标失调的后果。通过实时荧光定量PCR、免疫印迹、组织学、血糖-高胰岛素钳和糖耐量试验来确定后代脂肪组织中母体程序化miR-126-3p水平的表型和功能结局。蛋白质组学方法证实了miR-126-3p的已知靶点（包括IRS-1）的身份，并鉴定了内质网（ER）蛋白Lunapark作为一种新的靶点。我们通过荧光素酶测定证实Lunapark是miR-126-3p的直接靶点。在体外过表达miR-126-3p导致Lunapark蛋白水平降低，Perk（也称为Eif2ak3） mRNA水平升高，小干扰rna介导的Lunapark敲低导致Xbp1、剪接Xbp1、Chop（也称为Ddit3）和Perk mRNA水平升高，并在3T3-L1前脂肪细胞中产生内质网应激转录反应。与体外实验结果一致，肥胖母鼠所生的成年小鼠后代脂肪组织中miR-126-3p表达增加，同时伴有Lunapark和IRS-1蛋白水平下降，内质网应激标志物增加。在全身水平上，这些动物表现出葡萄糖耐受不良。同时靶向IRS-1和Lunapark，营养程序性地增加miR-126-3p导致脂肪组织胰岛素抵抗和内质网应激反应，这两者都可能导致糖耐量受损。这些发现提供了妊娠期肥胖导致后代2型糖尿病风险增加的新机制，因此确定了miR-126-3p作为潜在的治疗靶点。在线版本包含同行评审但未经编辑的补充材料，可在10.1007/s00125-020-05357-4获得。
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. 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. 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. 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. The online version contains peer-reviewed but unedited supplementary material available at 10.1007/s00125-020-05357-4.
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影响因子：
8.1
作者：
Fernandez-Twinn DS;Alfaradhi MZ;Martin-Gronert MS;Duque-Guimaraes DE;Piekarz A;Ferland-McCollough D;Bushell M;Ozanne SE
通讯作者：
Ozanne SE
影响因子：
4.8
作者：
Alfaradhi MZ;Kusinski LC;Fernandez-Twinn DS;Pantaleão LC;Carr SK;Ferland-McCollough D;Yeo GS;Bushell M;Ozanne SE
通讯作者：
Ozanne SE
影响因子：
7.7
作者：
McQuaid SE;Hodson L;Neville MJ;Dennis AL;Cheeseman J;Humphreys SM;Ruge T;Gilbert M;Fielding BA;Frayn KN;Karpe F
通讯作者：
Karpe F
影响因子：
8.2
作者：
Bickerton, A. S. T.;Roberts, R.;Frayn, K. N.
通讯作者：
Frayn, K. N.
DOI：
10.1073/pnas.1423026112
发表时间：
2015-01-13
影响因子：
11.1
作者：
Chen, Shuliang;Desai, Tanvi;Novick, Peter J.
通讯作者：
Novick, Peter J.