Flow-enhanced priming of hESCs through H2B acetylation and chromatin decondensation
Flow-enhanced priming of hESCs through H2B acetylation and chromatin decondensation
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通过 H2B 乙酰化和染色质解缩对 hESC 进行流动增强引发
DOI：
10.1186/s13287-019-1454-z
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发表时间：
2019-11
影响因子：
7.5
通讯作者：
Long M.
中科院分区：
文献类型：
--
作者：
Wang J.W.;Wu Y.;Zhang X.;Zhang F.;Lü D.Y.;Shangguan B.;Gao Y.X.;Long M.
背景已知不同的机械刺激可以操纵胚胎干细胞（embryonic stem cells，ESCs）的行为。ESC如何响应机械力和潜在的生物学效应的基本原理仍然难以捉摸。在这里，我们进行了mechanobiological研究hESC后mechanomics分析，以解开典型的mechanosensitive过程hESC-特定的流体shear.MethodshESC线H1进行系统地改变剪切流，和mechanosensitive蛋白质，通过质谱（MS）分析。然后，进行功能富集分析以鉴定富集的基因集。在1.1 Pa的稳定剪切流下24 h，使用蛋白质印迹（WB）、定量实时PCR（qPCR）和免疫荧光（IF）染色进一步检测蛋白质表达。同时，将细胞用200 nM阿司他汀（TSA）处理1小时作为阳性对照，以测试染色质去凝聚。然后分别用TRITC标记的鬼笔环肽、Hoechst 33342和SYTO® RNASelect™绿色荧光细胞染色剂（Life Technologies）使肌动蛋白、DNA和RNA可视化。此外，用原子力显微镜（AFM）测定细胞硬度，用流式细胞仪（FCM）测定膜联蛋白V-PE来测定细胞凋亡。结果在流体剪切力下的力学组学分析中，解开了体内hESCs相关的典型力学敏感蛋白。功能分析显示剪切流作用下hESC的组蛋白乙酰化、核大小和细胞骨架发生了显著改变。剪切流能够通过CFL 2/F-actin细胞骨架重组诱导H2 B乙酰化和核铺展。由此产生的染色质deconcensation和一个更大的细胞核很容易容纳信号分子和transcription factors.ConclusionsShear流调节染色质动力学在hESC通过细胞骨架和细胞核的改变，并巩固其启动状态。
BackgroundDistinct mechanical stimuli are known to manipulate the behaviors of embryonic stem cells (ESCs). Fundamental rationale of how ESCs respond to mechanical forces and the potential biological effects remain elusive. Here we conducted the mechanobiological study for hESCs upon mechanomics analysis to unravel typical mechanosensitive processes on hESC-specific fluid shear.MethodshESC line H1 was subjected to systematically varied shear flow, and mechanosensitive proteins were obtained by mass spectrometry (MS) analysis. Then, function enrichment analysis was performed to identify the enriched gene sets. Under a steady shear flow of 1.1 Pa for 24 h, protein expressions were further detected using western blotting (WB), quantitative real-time PCR (qPCR), and immunofluorescence (IF) staining. Meanwhile, the cells were treated with 200 nM trichostatin (TSA) for 1 h as positive control to test chromatin decondensation. Actin, DNA, and RNA were then visualized with TRITC-labeled phalloidin, Hoechst 33342, and SYTO® RNASelect™ green fluorescent cell stain (Life Technologies), respectively. In addition, cell stiffness was determined with atomic force microscopy (AFM) and annexin V-PE was used to determine the apoptosis with a flow cytometer (FCM).ResultsTypical mechanosensitive proteins were unraveled upon mechanomics analysis under fluid shear related to hESCs in vivo. Functional analyses revealed significant alterations in histone acetylation, nuclear size, and cytoskeleton for hESC under shear flow. Shear flow was able to induce H2B acetylation and nuclear spreading by CFL2/F-actin cytoskeletal reorganization. The resulting chromatin decondensation and a larger nucleus readily accommodate signaling molecules and transcription factors.ConclusionsShear flow regulated chromatin dynamics in hESCs via cytoskeleton and nucleus alterations and consolidated their primed state.
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影响因子：
3.5
作者：
Lüwen Zhou;Chen Zhang;Fan Zhang;Shouqin Lü;Shujin Sun;Dongyuan Lü;M. Long
通讯作者：
Lüwen Zhou;Chen Zhang;Fan Zhang;Shouqin Lü;Shujin Sun;Dongyuan Lü;M. Long
影响因子：
64.5
作者：
Percharde M;Lin CJ;Yin Y;Guan J;Peixoto GA;Bulut-Karslioglu A;Biechele S;Huang B;Shen X;Ramalho-Santos M
通讯作者：
Ramalho-Santos M
心脏的发育
影响因子：
--
作者：
G. Graham
通讯作者：
G. Graham
影响因子：
23.9
作者：
Nichols, Jennifer;Smith, Austin
通讯作者：
Smith, Austin
影响因子：
5.3
作者：
Li, Q;Lau, A;Stanley, EF
通讯作者：
Stanley, EF