Surface marker profiling of SH-SY5Y cells enables small molecule screens identifying BMP4 as a modulator of neuroblastoma differentiation.
Surface marker profiling of SH-SY5Y cells enables small molecule screens identifying BMP4 as a modulator of neuroblastoma differentiation.
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DOI：
10.1038/s41598-017-13497-8
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发表时间：
2017-10-19
影响因子：
4.6
通讯作者：
Pruszak J
中科院分区：
文献类型：
--
作者：
Ferlemann FC;Menon V;Condurat AL;Rößler J;Pruszak J
神经母细胞瘤是儿童最常见的颅外实体瘤。其广泛的临床结果反映了潜在的固有细胞异质性。由于目前的治疗通常不能根除肿瘤，因此需要更好地定义治疗抗性神经母细胞瘤并识别新的调节分子。为此，我们进行了第一次全面的流式细胞术表征表面分子表达的神经母细胞瘤细胞系。利用一个既定的聚类算法（SPADE）的细胞亚群的无偏可视化，我们进行了一个多孔筛选神经母细胞瘤表型的小分子调节剂。除SH-SY 5 Y细胞外，SH-EP、BE（2）-M17和Kelly细胞系也作为神经母细胞瘤的体外模型纳入随访分析。糖蛋白表位（CD 15、CD 24、CD 44、CD 57、TrkA）和趋化因子受体CXCR 4（CD 184）的组合检测使得能够定量鉴定对小分子有差异响应的SPADE定义的簇。研究发现，暴露于骨形态发生蛋白（BMP）-4可增强TrkAhigh/CD 15 −/CD 184 −神经母细胞瘤细胞亚群，同时伴有双皮质素阳性神经母细胞和SH-SY 5 Y细胞中NMYC蛋白表达的减少。除了产生用于研究神经母细胞瘤病理学的新的标记物候选物之外，我们的方法可以提供用于改进药理学筛选的工具，以开发神经母细胞瘤诊断和治疗的新途径。
Neuroblastoma is the most common extra-cranial solid tumor in children. Its broad spectrum of clinical outcomes reflects the underlying inherent cellular heterogeneity. As current treatments often do not lead to tumor eradication, there is a need to better define therapy-resistant neuroblastoma and to identify new modulatory molecules. To this end, we performed the first comprehensive flow cytometric characterization of surface molecule expression in neuroblastoma cell lines. Exploiting an established clustering algorithm (SPADE) for unbiased visualization of cellular subsets, we conducted a multiwell screen for small molecule modulators of neuroblastoma phenotype. In addition to SH-SY5Y cells, the SH-EP, BE(2)-M17 and Kelly lines were included in follow-up analysis as in vitro models of neuroblastoma. A combinatorial detection of glycoprotein epitopes (CD15, CD24, CD44, CD57, TrkA) and the chemokine receptor CXCR4 (CD184) enabled the quantitative identification of SPADE-defined clusters differentially responding to small molecules. Exposure to bone morphogenetic protein (BMP)-4 was found to enhance a TrkAhigh/CD15−/CD184− neuroblastoma cellular subset, accompanied by a reduction in doublecortin-positive neuroblasts and of NMYC protein expression in SH-SY5Y cells. Beyond yielding novel marker candidates for studying neuroblastoma pathology, our approach may provide tools for improved pharmacological screens towards developing novel avenues of neuroblastoma diagnosis and treatment.
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影响因子：
5.6
作者：
Ata R;Antonescu CN
通讯作者：
Antonescu CN
影响因子：
45.3
作者：
Combaret, V;Gross, N;Favrot, MC
通讯作者：
Favrot, MC
影响因子：
1
作者：
ERLANDSON, RA;NESLAND, JM
通讯作者：
NESLAND, JM
影响因子：
3.7
作者：
FLICKINGER, KS;JUDWARE, R;CULP, LA
通讯作者：
CULP, LA
影响因子：
0.4
作者：
Gao, Q.;Chen, C. F.;Zhang, H. Y.
通讯作者：
Zhang, H. Y.