Department of Biomedical Engineering, College of Engineering, The Ohio State University, Columbus, OH, United States of America.
The Ohio State University and Arthur G. James Comprehensive Cancer Center, Columbus, OH, United States of America.
Biofabrication. 2024 Oct 24;17(1). doi: 10.1088/1758-5090/ad82e0.
Progressive metastasis is the primary cause of cancer-related deaths. It has been recognized that many cancers are characterized by long periods of stability followed by subsequent progression. Genes termed metastasis progression suppressors (MPS) are functional gatekeepers of this process, and their loss leads to late-stage progression. Previously, we identified regulator of calcineurin 1, isoform 4 (RCAN1.4) as a functional MPS for several cancers, including thyroid cancer, a tumor type prone to metastatic dormancy. RCAN1.4 knockdown increases expression of the cancer-promoting transcription factor NFE2-like bZIP transcription factor (NFE2L3), and through this mechanism increases cancer cell proliferation and invasion inandand promotes metastatic potential to lungs in tail vein models. However, the mechanisms by which RCAN 1.4 regulates specific metastatic steps is incompletely characterized. Studies of the metastatic cascade are limited in mouse systems due to high cost and long duration. Here, we have shown the creation of a thyroid-to-lung metastasis-on-a-chip (MOC) model to address these limitations, allowing invasion analysis and quantification on a single cell level. We then deployed the platform to investigate RCAN1.4 knockdown in fluorescently tagged hTh74 and FTC236 thyroid cancer cell lines. Cells were circulated through microfluidic channels, running parallel to lung hydrogel constructs allowing tumor cell-lung tissue interactions. Similar to studies in mouse models, RCAN1.4 knockdown increased NFE2L3 expression, globally increased invasion distance into lung constructs and had cell line and clonally dependent variations on bulk metastatic burden. In line with previousobservations, RCAN1.4 knockdown had a greater impact on hTh74 metastatic propensity than FTC236. In summary, we have developed and validated a novel MOC system evaluate and quantify RCAN1.4-regulated thyroid cancer cell lung adherence and invasion. This system creates opportunities for more detailed and rapid mechanistic studies the metastatic cascade and creates opportunities for translational assay development.
进行性转移是癌症相关死亡的主要原因。人们已经认识到，许多癌症的特征是在随后的进展之前存在很长一段时间的稳定期。被称为转移进展抑制剂（MPS）的基因是这个过程的功能守门员，它们的丢失导致晚期进展。以前，我们确定钙调神经磷酸酶 1，同种型 4（RCAN1.4）为几种癌症（包括甲状腺癌，一种易于发生转移休眠的肿瘤类型）的功能性 MPS。RCAN1.4 敲低会增加促进癌症的转录因子 NFE2 样 bZIP 转录因子（NFE2L3）的表达，并且通过这种机制增加了癌细胞在体内的增殖和侵袭，并促进了尾静脉模型中肺部的转移潜力。然而，RCAN1.4 调节特定转移步骤的机制尚未完全阐明。由于成本高和持续时间长，在小鼠系统中对转移级联的研究受到限制。在这里，我们展示了创建甲状腺到肺转移芯片（MOC）模型来解决这些限制，允许在单细胞水平上进行侵袭分析和量化。然后，我们将该平台用于研究荧光标记的 hTh74 和 FTC236 甲状腺癌细胞系中 RCAN1.4 的敲低。细胞在微流控通道中循环，与肺水凝胶结构平行运行，允许肿瘤细胞-肺组织相互作用。与小鼠模型中的研究相似，RCAN1.4 敲低增加了 NFE2L3 的表达，总体上增加了进入肺结构的侵袭距离，并对批量转移负担具有细胞系和克隆依赖性变化。与之前的观察结果一致，RCAN1.4 敲低对 hTh74 的转移倾向的影响大于 FTC236。总之，我们已经开发并验证了一种新型 MOC 系统来评估和量化 RCAN1.4 调节的甲状腺癌细胞对肺的粘附和侵袭。该系统为更详细和快速的转移级联机制研究创造了机会，并为转化测定的发展创造了机会。
