Endometrium-on-a-chip reveals the endometrial transcriptome, and protein content of secretome are altered by changes in circulating concentrations of insulin and glucose in vitro
Endometrium-on-a-chip reveals the endometrial transcriptome, and protein content of secretome are altered by changes in circulating concentrations of insulin and glucose in vitro
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子宫内膜芯片揭示子宫内膜转录组，分泌组的蛋白质含量会因体外胰岛素和葡萄糖循环浓度的变化而改变
DOI：
10.1101/2020.11.03.361774
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
De Bem T
中科院分区：
文献类型：
--
作者：
De Bem T
母体环境和发育中的胚胎之间的分子相互作用是早期妊娠成功的关键，已知会受到代谢状态等因素的影响。然而，我们对这些个体营养应激改变子宫内膜功能和子宫内环境以促进早期妊娠成功的机制的理解有限。在这里，我们首次报道了使用芯片上子宫内膜微流体方法在体外产生多细胞子宫内膜，该子宫内膜暴露于与母体代谢应激相关的葡萄糖和胰岛素浓度。在从处于黄体早期（大约第4-7天）的非妊娠母牛的子宫中分离子宫内膜细胞（上皮细胞和基质细胞）后，将上皮细胞接种到上室（4-6 × 104个细胞/mL）中，并将基质细胞接种到下室（1.5-2 × 104个细胞/mL）中。三种不同浓度的葡萄糖1）0.5 mM 2）5.0 mM或3）50 mM或胰岛素1）媒介物，2）1 ng/mL或3）10 ng/mL在子宫内膜细胞中以1µL/min的流速进行72小时，以模拟体内分泌速率。通过RNA测序和TMT分别测定细胞转录组反应和体外子宫腔液（ULF）分泌蛋白质组的定量差异。母体葡萄糖的变化分别改变了上皮细胞和间质细胞中21个和191个蛋白编码基因（p<0.05）。而蛋白分泌组（1和23蛋白）则呈剂量依赖性变化。胰岛素导致有限的转录变化，包括细胞特异性的胰岛素样结合蛋白（5、12和20），但改变了196种蛋白质的定量分泌，包括参与细胞外基质-受体相互作用和癌症中蛋白聚糖信号传导的蛋白质。总的来说，这些都突出了母体葡萄糖和胰岛素改变子宫功能的潜在机制。
The molecular interactions between the maternal environment and developing embryo that are key for early pregnancy success are known to be influenced by factors such as the metabolic status. We are, however, limited in our understanding of the mechanism by which these individual nutritional stressors alter endometrial function and thein uteroenvironment for early pregnancy success. Here we report for the first time the use of endometrium-on-a-chip microfluidics approach to produce a multi-cellular endometriumin vitro, that is exposed to glucose and insulin concentrations associated with maternal metabolic stressors. Following isolation of endometrial cells (epithelial and stromal) from the uteri of non-pregnant cows in early-luteal phase (Day 4-7 approximately) epithelial cells were seeded into the upper chamber (4-6 104cells/mL) and stromal cells seeded in the lower chamber (1.5-2 104cells/mL). Three different concentration of glucose 1) 0.5 mM 2) 5.0 mM or 3) 50 mM or insulin 1) Vehicle, 2) 1 ng/mL or 3) 10 ng/mL were performed in the endometrial cells at a flow rate of 1µL/min for 72 hr to mimic the rate of secretionin vivo. Quantitative differences in the transcriptomic response of the cells and the secreted proteome ofin vitro-derived uterine luminal fluid (ULF) were determined by RNA-sequencing andTMTrespectively. Changes in maternal glucose altered 21 and 191 protein coding genes in epithelial and stromal cells respectively (p<0.05). While there was a dose-dependent quantitative change in protein secretome (1 and 23 proteins). Insulin resulted in limited transcriptional changes including insulin-like binding proteins that were cell specific (5, 12, and 20) but altered the quantitative secretion of 196 proteins including those involved in extracellular matrix-receptor interaction and proteoglycan signaling in cancer. Collectively, these highlight the potential mechanism by which changes to maternal glucose and insulin alter uterine function.
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影响因子：
3.8
作者：
C. Passaro;D. Tutt;Sandra Bages Arnal;C. Maicas;R. Laguna;A. Gutiérrez;J. Browne;D. Rath;S. Behura;T. Spencer;T. Fair;P. Lonergan
通讯作者：
P. Lonergan
影响因子：
4.8
作者：
Yang, ZZ;Tschopp, O;Hemmings, BA
通讯作者：
Hemmings, BA
荧光法检测发情周期不同阶段和妊娠早期牛输卵管上皮和基质细胞以及子宫内膜基质细胞中血小板激活因子受体的含量。
影响因子：
2.1
作者：
U. Tiemann;T. Viergutz;Ludwig Jonas;K. Wollenhaupt;R. Pöhland;W. Kanitz
通讯作者：
W. Kanitz
动情周期和妊娠早期小鼠卵巢和输卵管中血小板激活因子受体的定位和分布
影响因子：
3.6
作者：
G. Lash;M. Legge
通讯作者：
M. Legge
子宫内膜外植体的三维体外培养模拟子宫内膜异位症的早期阶段。
影响因子：
6.7
作者：
A. Fasciani;G. Bocci;Jing Xu;R. Bielecki;E. Greenblatt;N. Leyland;R. Casper
通讯作者：
R. Casper