Activation of the Hippo/TAZ pathway is required for menstrual stem cells to suppress myofibroblast and inhibit transforming growth factor signaling in human endometrial stromal cells
Activation of the Hippo/TAZ pathway is required for menstrual stem cells to suppress myofibroblast and inhibit transforming growth factor signaling in human endometrial stromal cells
复制标题
月经干细胞需要激活 Hippo/TAZ 通路来抑制肌成纤维细胞并抑制人子宫内膜基质细胞中的转化生长因子信号传导
DOI：
10.1093/humrep/dez001
复制
发表时间：
2019-02
影响因子：
6.1
通讯作者：
Zhang Songying
中科院分区：
文献类型：
--
作者：
Zhu Haiyan;Pan Yibin;Jiang Yinshen;Li Jing;Zhang Yanling;Zhang Songying
研究问题
月经干细胞（MenSC）能否抑制人子宫内膜间质细胞（ESCs）中肌成纤维细胞分化和逆转转化生长因子β（TGFβ）介导的肌成纤维细胞表型活化？
答复摘要
MenSC抑制子宫内膜肌成纤维细胞分化并逆转TGFβ介导的肌成纤维细胞表型活化，这可能与Hippo/TAZ通路的活化相关。
已知的
MenSC作为细胞疗法的潜在作用包括减弱子宫内粘连，但MenSC发挥这些作用的潜在机制尚未完全了解。
研究设计、规模、持续时间
我们评估了MenSC对肌成纤维细胞分化的拮抗作用以及Hippo/TAZ信号传导途径对TGFβ介导的肌成纤维细胞基因表达诱导的更广泛作用。研究设计基于从三名月经周期规律的健康绝经前女性获得的临床增殖期子宫内膜样本的队列。
受试者/材料、环境、方法
将ESC与MenSC共培养或在MenSC条件培养基中共培养。评价纤维化标志物（αSMA、胶原I、CTGF和纤连蛋白）以及增殖和伤口愈合能力。还研究了Hippo/TAZ通路的组分（TAZ、p-TAZ、MOB 1、p-MOB 1、LATS 1和p-LATS 1）。细胞计数试剂盒8，伤口愈合试验，实时PCR，免疫印迹，免疫荧光和shRNA敲低方法来验证的结果。
主要结果和机会的作用
MenSC抑制肌成纤维细胞活化，导致ESC更快速增殖。MenSC下调肌成纤维细胞标志物αSMA和胶原I的表达，促进子宫内膜伤口愈合。与MenSC共培养还减弱了TGFβ介导的纤维化标记基因αSMA、胶原I、CTGF和纤连蛋白表达的增加，并恢复了TGFβ抑制的伤口愈合能力。MenSC诱导Hippo/TAZ通路活化，导致TAZ的核输出和细胞质保留。TAZ抑制被证明即使在不存在MenSC的情况下也具有类似的效果，并且TAZ的抑制足以减弱TGFβ介导的肌成纤维细胞活化。
大规模数据
N/A.
限制、豁免理由
本研究仅包括体外实验。因此，在未来的研究中需要来自体内实验的额外数据。
调查结果的更广泛影响
Hippo/TAZ通路可能是子宫内膜纤维化的重要治疗靶点。
研究资金/竞争性兴趣
本研究得到了国家自然科学基金项目（No.81601236）和浙江省自然科学基金项目（LY 19 H 040009）的资助。没有一位作者有任何利益冲突需要声明。
STUDY QUESTION
Can menstrual stem cells (MenSCs) inhibit myofibroblast differentiation and reverse transforming growth factor β (TGFβ)-mediated activation of myofibroblast phenotypes in human endometrial stromal cells (ESCs)?
SUMMARY ANSWER
MenSCs suppressed endometrial myofibroblast differentiation and reversed TGFβ-mediated activation of myofibroblast phenotypes, which might be associated with activation of the Hippo/TAZ pathway.
WHAT IS KNOWN ALREADY
The potential effect of MenSCs as a cell therapy include attenuation of intrauterine adhesions, but the underlying mechanisms by which MenSCs exerts these effects are not entirely understood.
STUDY DESIGN, SIZE, DURATION
We evaluated the antagonistic effects of MenSCs on myofibroblast differentiation as well as the broader effect of the Hippo/TAZ signaling pathway on TGFβ-mediated induction of myofibroblast gene expression. The study design was based on a cohort of clinical proliferative phase endometrial samples obtained from three healthy premenopausal females with regular menstrual cycles.
PARTICIPANTS/MATERIALS, SETTING, METHODS
ESCs were cocultured with MenSCs or in MenSC-conditioned medium. Fibrotic markers (αSMA, collagen I, CTGF and fibronectin) as well as proliferation and wound-healing abilities were evaluated. Components of the Hippo/TAZ pathway (TAZ, p-TAZ, MOB1, p-MOB1, LATS1 and p-LATS1) were also investigated. Cell Counting Kit 8, wound healing assay, real-time PCR, western blotting, immunofluorescence and shRNA knockdown approaches were used to validate the findings.
MAIN RESULTS AND THE ROLE OF CHANCE
MenSCs inhibited myofibroblast activation, resulting in more rapid proliferation of ESCs. MenSCs downregulated the expression of myofibroblast markers αSMA and collagen I and promoted endometrial wound healing. Coculture with MenSCs also attenuated the TGFβ-mediated increase in expression of fibrotic marker genes αSMA, collagen I, CTGF and fibronectin, and restored the wound-healing ability inhibited by TGFβ. MenSCs induced Hippo/TAZ pathway activation, resulting in nuclear export and cytoplasmic retention of TAZ. TAZ inhibition was demonstrated to have similar effects even in the absence of MenSCs, and inhibition of TAZ was sufficient to attenuate TGFβ-mediated myofibroblast activation.
LARGE SCALE DATA
N/A.
LIMITATIONS, REASONS FOR CAUTION
This study included only in vitro experiments. Thus, additional data from in vivo experiments are needed in a future study.
WIDER IMPLICATIONS OF THE FINDINGS
The Hippo/TAZ pathway may be an important therapeutic target for endometrial fibrosis.
STUDY FUNDING/COMPETING INTEREST(S)
This study was supported by the National Natural Science Foundation of China (No. 81601236) and Zhejiang Provincial Natural Science Foundation of China (LY19H040009). None of the authors has any competing interests to declare.
登录
查看更多内容
影响因子：
6
作者：
Chen L;Zhang C;Chen L;Wang X;Xiang B;Wu X;Guo Y;Mou X;Yuan L;Chen B;Wang J;Xiang C
通讯作者：
Xiang C
影响因子：
--
作者：
Schüring AN;Braun J;Wüllner S;Kiesel L;Götte M
通讯作者：
Götte M
影响因子：
6.1
作者：
Haiyan Zhu;Jun-xia Wang;X. Tong;Wei-hai Xu;Ling-ying Jiang;Xiao-ying Jing;Ling-yun Yang;F. Zhou;Song-Ying Zhang
通讯作者：
Haiyan Zhu;Jun-xia Wang;X. Tong;Wei-hai Xu;Ling-ying Jiang;Xiao-ying Jing;Ling-yun Yang;F. Zhou;Song-Ying Zhang
影响因子：
3.9
作者：
Piersma B;Bank RA;Boersema M
通讯作者：
Boersema M
影响因子：
--
作者：
Singh N;Mohanty S;Seth T;Shankar M;Bhaskaran S;Dharmendra S
通讯作者：
Dharmendra S