Three-Dimensional Microtumors for Probing Heterogeneity of Invasive Bladder Cancer
Three-Dimensional Microtumors for Probing Heterogeneity of Invasive Bladder Cancer
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DOI：
10.1021/acs.analchem.0c00057
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发表时间：
2020-07-07
影响因子：
7.4
通讯作者：
Wong, Pak Kin
中科院分区：
文献类型：
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作者：
Torab, Peter;Yan, Yue;Wong, Pak Kin
膀胱癌是一种越来越常见的恶性肿瘤，肌肉浸润性膀胱癌的发病率和死亡率特别高。膀胱癌的形态和分子多样性对阐明导致疾病进展的侵袭机制提出了重大挑战。此外，传统的侵袭测定不能提供用于研究 3D 微环境中膀胱癌侵袭的生理背景，并且捕获细胞表型异质性对疾病进展的贡献的能力有限。在这里，我们描述了适用于分析膀胱癌患者细胞系和原发性肿瘤细胞的细胞表型异质性的 3D 微肿瘤侵袭模型的开发。该模型采用自组装方法，可在 3D 微环境中重现膀胱癌侵袭的特征并探测侵袭性细胞亚群。通过结合金纳米棒锁定核酸生物传感器来分析入侵微肿瘤的基因表达谱。将单细胞生物传感器和瞬时基因敲低纳入系统揭示了 Delta 样配体 4 (DLL4) 表达上调的侵袭性前导细胞的形成，以及 NOTCH1-DLL4 信号在集体膀胱癌侵袭中的作用。在经尿道切除术获得的患者样本中也观察到表达 DLL4 的细胞参与膀胱癌侵袭。总的来说，我们的研究展示了一种 3D 微肿瘤侵袭模型，用于研究膀胱癌侵袭的细胞内异质性并分析患者来源的样本以实现个性化医疗应用。
Bladder cancer is an increasingly common malignancy, and muscle invasive bladder cancer is associated with particularly high rates of morbidity and mortality. The morpho- logic and molecular diversity of bladder cancer poses significant challenges in elucidating the invasion mechanisms responsible for disease progression. Furthermore, conventional invasion assays do not provide a physiological context for studying bladder cancer invasion within 3D microenvironments and have limited ability to capture the contribution of cellular phenotypic heterogeneity to disease progression. Here, we describe the development of a 3D microtumor invasion model suitable for the analysis of cellular phenotypic heterogeneity in cell lines and primary tumor cells from bladder cancer patients. This model incorporates a self-assembly approach for recapitulating features of bladder cancer invasion in 3D microenvironments and probing the invasive cell subpopulations. The gene expression profiles of invading microtumors were analyzed by incorporating a gold nanorod-locked nucleic acid biosensor. The incorporation of the single cell biosensor and transient gene knockdown into the system revealed the formation of invasive leader cells with upregulated Delta-like ligand 4 (DLL4) expression as well as the role of NOTCH1-DLL4 signaling in collective bladder cancer invasion. The involvement of DLL4 expressing cells in bladder cancer invasion was also observed in patient samples obtained from transurethral resection. Collectively, our study demonstrates a 3D microtumor invasion model for investigating intracellular heterogeneity of bladder cancer invasion and analyzing patient derived samples toward personalized medicine applications.