Functional Isolation of Tumor-Initiating Cells using Microfluidic-Based Migration Identifies Phosphatidylserine Decarboxylase as a Key Regulator.
Functional Isolation of Tumor-Initiating Cells using Microfluidic-Based Migration Identifies Phosphatidylserine Decarboxylase as a Key Regulator.
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DOI：
10.1038/s41598-017-18610-5
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发表时间：
2018-01-10
影响因子：
4.6
通讯作者：
Yoon E
中科院分区：
文献类型：
--
作者：
Chen YC;Humphries B;Brien R;Gibbons AE;Chen YT;Qyli T;Haley HR;Pirone ME;Chiang B;Xiao A;Cheng YH;Luan Y;Zhang Z;Cong J;Luker KE;Luker GD;Yoon E
肿瘤起始细胞的分离目前依赖于不反映这些细胞的基本生物学功能的标记物。我们提出通过基于增强的迁移分离肿瘤起始细胞来克服这一限制，增强的迁移是通过上皮细胞向间质细胞转化（EMT）与肿瘤起始潜能紧密相关的功能。我们开发了一种高通量微流体迁移平台，该平台具有自动化细胞跟踪软件和用于下游功能和遗传分析的细胞的简易回收。使用这种装置，我们分离出一小部分迁移细胞，在小鼠模型中具有显著更大的肿瘤形成和转移。对来自两个转移性乳腺癌细胞系的迁移细胞与非迁移细胞的全转录组测序揭示了一组独特的基因作为肿瘤起始细胞的关键调节因子。我们专注于磷脂酰丝氨酸脱羧酶（PISD），一个在迁移细胞中下调8倍的基因。在高通量微流控乳腺微球装置和小鼠异种移植模型中，过表达PISD的乳腺癌细胞表现出降低的肿瘤引发潜力。PISD调节线粒体的多个方面，突出了线粒体作为癌症干细胞治疗靶点的功能。这项研究不仅建立了一种新的微流控技术，用于功能性分离肿瘤起始细胞，无论癌症类型如何，而且还建立了一种新的方法来识别这些细胞的基本调节因子作为药物开发的靶点。
Isolation of tumor-initiating cells currently relies on markers that do not reflect essential biologic functions of these cells. We proposed to overcome this limitation by isolating tumor-initiating cells based on enhanced migration, a function tightly linked to tumor-initiating potential through epithelial-to-mesenchymal transition (EMT). We developed a high-throughput microfluidic migration platform with automated cell tracking software and facile recovery of cells for downstream functional and genetic analyses. Using this device, we isolated a small subpopulation of migratory cells with significantly greater tumor formation and metastasis in mouse models. Whole transcriptome sequencing of migratory versus non-migratory cells from two metastatic breast cancer cell lines revealed a unique set of genes as key regulators of tumor-initiating cells. We focused on phosphatidylserine decarboxylase (PISD), a gene downregulated by 8-fold in migratory cells. Breast cancer cells overexpressing PISD exhibited reduced tumor-initiating potential in a high-throughput microfluidic mammosphere device and mouse xenograft model. PISD regulated multiple aspects of mitochondria, highlighting mitochondrial functions as therapeutic targets against cancer stem cells. This research establishes not only a novel microfluidic technology for functional isolation of tumor-initiating cells regardless of cancer type, but also a new approach to identify essential regulators of these cells as targets for drug development.
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影响因子：
50.3
作者：
Chen, Ruihuan;Nishimura, Merry C.;Phillips, Heidi S.
通讯作者：
Phillips, Heidi S.
影响因子：
3.9
作者：
Hellmich, W;Regtmeier, J;Ros, A
通讯作者：
Ros, A
影响因子：
23.9
作者：
Brooks MD;Burness ML;Wicha MS
通讯作者：
Wicha MS
影响因子：
6.1
作者：
Cheng YH;Chen YC;Brien R;Yoon E
通讯作者：
Yoon E
影响因子：
5.9
作者：
Liu S;Cong Y;Wang D;Sun Y;Deng L;Liu Y;Martin-Trevino R;Shang L;McDermott SP;Landis MD;Hong S;Adams A;D'Angelo R;Ginestier C;Charafe-Jauffret E;Clouthier SG;Birnbaum D;Wong ST;Zhan M;Chang JC;Wicha MS
通讯作者：
Wicha MS