Targeting ectodysplasin promotor by CRISPR/dCas9-effector effectively induces the reprogramming of human bone marrow-derived mesenchymal stem cells into sweat gland-like cells.
Targeting ectodysplasin promotor by CRISPR/dCas9-effector effectively induces the reprogramming of human bone marrow-derived mesenchymal stem cells into sweat gland-like cells.
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通过 CRISPR/dCas9 效应子靶向外胚增生素 (EDA) 启动子，有效诱导人骨髓来源的间充质干细胞重编程为汗腺样细胞
DOI：
10.1186/s13287-017-0758-0
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发表时间：
2018-01-12
影响因子：
7.5
通讯作者：
Fu X
中科院分区：
文献类型：
--
作者：
Sun S;Xiao J;Huo J;Geng Z;Ma K;Sun X;Fu X
深度烧伤患者的特点是失去排汗功能，无法再生汗腺。骨髓间充质干细胞(BM-MSCs)具有易获得、多潜能、免疫原性低等特点，是一种理想的细胞治疗生物来源。本研究的目的是确定以CRISPR/dCas9-Effect(dCas9-E)为靶点的胞外异位基因启动子(EDA)能否诱导BM-MSCs分化为汗腺样细胞(SGCs)。将Lenti-CRISPR/dCas9-效应子和单引导RNA(SgRNAs)导入BM-MSCs，激活EDA转录。在体外和小鼠模型中鉴定和评价了dCas9-E BM-MSCs对SGCs形成和烧伤修复的影响。在多西环素(Dox)诱导下，sgRNA引导的dCas9-E转染BM-MSCs后，细胞内EDA的转录和表达显著增加。有趣的是，在BM-MSCs中汗腺的特异性标志物(CEA、CK7、CK14和CK19)也呈阳性，提示EDA在促进BM-MSC向汗腺分化中起关键作用。此外，将经Dox诱导的dCas9-E BM-MSCs植入实验动物模型的创面，碘淀粉排汗试验显示处理后的爪子出汗阳性，而生理盐水处理的爪子则呈阴性表现。在调控机制上，NF-κB下游基因(Shh和Cyclin D1)的表达也相应增强。这些结果表明，EDA是BM-MSCs汗腺再生的关键因素，也可能为汗腺破坏和大面积深度烧伤提供一种新的方法。本文的在线版本(doi：10.1186/s13287-0170758-0)包含补充材料，授权用户可以使用。
Patients with a deep burn injury are characterized by losing the function of perspiration and being unable to regenerate the sweat glands. Because of their easy accession, multipotency, and lower immunogenicity, bone marrow-derived mesenchymal stem cells (BM-MSCs) represent as an ideal biological source for cell therapy. The aim of this study was to identify whether targeting the promotor of ectodysplasin (EDA) by CRISPR/dCas9-effector (dCas9-E) could induce the BM-MSCs to differentiate into sweat gland-like cells (SGCs). Activation of EDA transcription in BM-MSCs was attained by transfection of naive BM-MSCs with the lenti-CRISPR/dCas9-effector and single-guide RNAs (sgRNAs). The impact of dCas9-E BM-MSCs on the formation of SGCs and repair of burn injury was identified and evaluated both in vitro and in a mouse model. After transfection with sgRNA-guided dCas9-E, the BM-MSCs acquired significantly higher transcription and expression of EDA by doxycycline (Dox) induction. Intriguingly, the specific markers (CEA, CK7, CK14, and CK19) of sweat glands were also positive in the transfected BM-MSCs, suggesting that EDA plays a critical role in promoting BM-MSC differentiation into sweat glands. Furthermore, when the dCas9-E BM-MSCs with Dox induction were implanted into a wound in a laboratory animal model, iodine-starch perspiration tests revealed that the treated paws were positive for perspiration, while the paws treated with saline showed a negative manifestation. For the regulatory mechanism, the expression of downstream genes of NF-κB (Shh and cyclin D1) was also enhanced accordingly. These results suggest that EDA is a pivotal factor for sweat gland regeneration from BM-MSCs and may also offer a new approach for destroyed sweat glands and extensive deep burns. The online version of this article (doi:10.1186/s13287-017-0758-0) contains supplementary material, which is available to authorized users.
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影响因子：
3.5
作者：
Pispa, Johanna;Pummila, Marja;Mikkola, Marja L.
通讯作者：
Mikkola, Marja L.
影响因子：
4.6
作者：
Schmidt-Ullrich, R;Tobin, DJ;Scheidereit, C
通讯作者：
Scheidereit, C
影响因子：
1.7
作者：
Li, Hai-Hong;Zhou, Gang;Zhang, Lei
通讯作者：
Zhang, Lei
DOI：
10.1126/science.1232033
发表时间：
2013-02-15
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Mali P;Yang L;Esvelt KM;Aach J;Guell M;DiCarlo JE;Norville JE;Church GM
通讯作者：
Church GM
影响因子：
5.4
作者：
Gao Y;Li M;Zhang X;Bai T;Chi G;Liu JY;Li Y
通讯作者：
Li Y