Molecular Characterization of Differentiated-Resistance MSC Subclones by Single-Cell Transcriptomes.
Molecular Characterization of Differentiated-Resistance MSC Subclones by Single-Cell Transcriptomes.
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DOI：
10.3389/fcell.2022.699144
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发表时间：
2022
影响因子：
5.5
通讯作者：
Zhong JF
中科院分区：
文献类型：
--
作者：
Stucky A;Gao L;Li SC;Tu L;Luo J;Huang X;Chen X;Li X;Park TH;Cai J;Kabeer MH;Plant AS;Sun L;Zhang X;Zhong JF
背景资料：间充质干细胞（MSCs）潜在的致瘤性机制仍不清楚，导致临床应用效果不一致。我们假设MSC中的亚克隆有助于其致瘤性，并且我们在单细胞水平上接近MSC亚克隆。 研究方法：将MSC在成骨分化培养基中培养，并在第12、19和25天收获，用于使用茜素红的细胞分化分析，随后进行单细胞转录组。 结果如下：单细胞RNA-seq分析揭示了成骨过程中MSC的离散簇，包括分化抗性MSC（DR-MSC）、分化的成骨细胞（DO）和前体成骨细胞（PO）。DR-MSC群体类似于癌症起始细胞，并对yes相关蛋白1（YAP 1）网络进行进一步分析。维替泊芬还用于癌细胞系中的YAP 1抑制，以证实YAP 1在MSC参与的致瘤性中的作用。分析了各种癌症类型的临床数据，以揭示YAP 1、OCT 4和CDH 6在MSC参与的致瘤性中的关系。与PO和DO相比，DR-MSCs中钙粘蛋白6（CDH 6）、八聚体结合转录因子4（OCT 4）和YAP 1的表达显著上调。维替泊芬抑制YAP 1可加速MSCs的分化，并抑制YAP 1、CDH 6和OCT 4的表达。对56个临床队列的调查显示，在各种实体瘤中，CDH 6、YAP 1和OCT 4之间存在高度的共表达。YAP 1抑制也下调HeLa细胞活力，并逐渐抑制YAP 1核定位，同时减少CDH 6和OCT 4的转录。 结论：我们使用单细胞测序来分析未分化的MSC，并发现分化抗性亚克隆的单细胞MSC中的致癌途径。
Background: The mechanism of tumorigenicity potentially evolved in mesenchymal stem cells (MSCs) remains elusive, resulting in inconsistent clinical application efficacy. We hypothesized that subclones in MSCs contribute to their tumorgenicity, and we approached MSC-subclones at the single-cell level. Methods: MSCs were cultured in an osteogenic differentiation medium and harvested on days 12, 19, and 25 for cell differentiation analysis using Alizarin Red and followed with the single-cell transcriptome. Results: Single-cell RNA-seq analysis reveals a discrete cluster of MSCs during osteogenesis, including differentiation-resistant MSCs (DR-MSCs), differentiated osteoblasts (DO), and precursor osteoblasts (PO). The DR-MSCs population resembled cancer initiation cells and were subjected to further analysis of the yes associated protein 1 (YAP1) network. Verteporfin was also used for YAP1 inhibition in cancer cell lines to confirm the role of YAP1 in MSC--involved tumorigenicity. Clinical data from various cancer types were analyzed to reveal relationships among YAP1, OCT4, and CDH6 in MSC--involved tumorigenicity. The expression of cadherin 6 (CDH6), octamer-binding transcription factor 4 (OCT4), and YAP1 expression was significantly upregulated in DR-MSCs compared to PO and DO. YAP1 inhibition by Verteporfin accelerated the differentiation of MSCs and suppressed the expression of YAP1, CDH6, and OCT4. A survey of 56 clinical cohorts revealed a high degree of co-expression among CDH6, YAP1, and OCT4 in various solid tumors. YAP1 inhibition also down-regulated HeLa cell viability and gradually inhibited YAP1 nuclear localization while reducing the transcription of CDH6 and OCT4. Conclusions: We used single-cell sequencing to analyze undifferentiated MSCs and to discover a carcinogenic pathway in single-cell MSCs of differentiated resistance subclones.
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影响因子：
24.5
作者：
Deng N;Goh LK;Wang H;Das K;Tao J;Tan IB;Zhang S;Lee M;Wu J;Lim KH;Lei Z;Goh G;Lim QY;Tan AL;Sin Poh DY;Riahi S;Bell S;Shi MM;Linnartz R;Zhu F;Yeoh KG;Toh HC;Yong WP;Cheong HC;Rha SY;Boussioutas A;Grabsch H;Rozen S;Tan P
通讯作者：
Tan P
影响因子：
21.3
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影响因子：
2.9
作者：
Kim RJ;Nam JS
通讯作者：
Nam JS
影响因子：
3.7
作者：
Chopra, Martin;Bohlander, Stefan K.
通讯作者：
Bohlander, Stefan K.
影响因子：
--
作者：
Li SC;Stucky A;Chen X;Kabeer MH;Loudon WG;Plant AS;Torno L;Nangia CS;Cai J;Zhang G;Zhong JF
通讯作者：
Zhong JF