A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity.
A 3D adrenocortical carcinoma tumor platform for preclinical modeling of drug response and matrix metalloproteinase activity.
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DOI：
10.1038/s41598-023-42659-0
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发表时间：
2023-09-19
影响因子：
4.6
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中科院分区：
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肾上腺皮质癌（ACC）预后较差，几十年来一直没有发现新药。药物开发的缺乏部分归因于缺乏临床前模型。 ACC 的动物模型和 2D 细胞培养物都无法准确模拟该疾病，因为动物生理学本质上与人类不同，并且 2D 培养物无法代表关键的 3D 结构。类器官和其他小型组织或肿瘤 3D 体外模型可以模拟人类体内生物学的某些复杂性；然而，这项技术在很大程度上尚未应用于 ACC。在这项研究中，我们描述了从已建立的 ACC 细胞系 NCI-H295R 生成 3D 肿瘤构建体。 NCI-H295R 细胞被封装以生成 3D ACC 构建体。评估肿瘤构建体的生物标志物表达、活力、增殖和皮质醇产生。此外，使用荧光 MMP 敏感生物传感器并通过将 NCI-H295R 细胞输注到片上转移微流体装置平台中，直接评估基质金属蛋白酶 (MMP) 功能。 ACC 肿瘤构建体显示与 ACC 相关的生物标志物的表达，包括 SF-1、Melan A 和抑制素 α。与 2D 细胞培养相比，用化疗药物治疗 ACC 肿瘤构建体的药物敏感性降低。由于大多数肿瘤细胞使用 MMP 分解细胞外基质在组织中迁移，因此我们通过在肿瘤构建体中集成荧光 MMP 敏感肽生物传感器来验证 ACC 肿瘤构建体的实用性。最后，在我们的芯片转移装置中，NCI-H295R 细胞成功移植到基于下游肺细胞系的构建体中，但进入肺构建体的侵袭距离因 MMP 抑制而缩短。这些使用 2D 细胞培养物不可能实现的研究表明，NCI-H295R 细胞分泌用于 3D 侵袭的活性 MMP。这项工作代表了 ACC 3D 肿瘤构建平台的第一个证据，该平台可用于未来的机制研究以及干预和治疗新靶点的开发。
Adrenocortical carcinoma (ACC) has a poor prognosis, and no new drugs have been identified in decades. The absence of drug development can partly be attributed to a lack of preclinical models. Both animal models and 2D cell cultures of ACC fail to accurately mimic the disease, as animal physiology is inherently different than humans, and 2D cultures fail to represent the crucial 3D architecture. Organoids and other small 3D in vitro models of tissues or tumors can model certain complexities of human in vivo biology; however, this technology has largely yet to be applied to ACC. In this study, we describe the generation of 3D tumor constructs from an established ACC cell line, NCI-H295R. NCI-H295R cells were encapsulated to generate 3D ACC constructs. Tumor constructs were assessed for biomarker expression, viability, proliferation, and cortisol production. In addition, matrix metalloproteinase (MMP) functionality was assessed directly using fluorogenic MMP-sensitive biosensors and through infusion of NCI-H295R cells into a metastasis-on-a-chip microfluidic device platform. ACC tumor constructs showed expression of biomarkers associated with ACC, including SF-1, Melan A, and inhibin α. Treatment of ACC tumor constructs with chemotherapeutics demonstrated decreased drug sensitivity compared to 2D cell culture. Since most tumor cells migrate through tissue using MMPs to break down extracellular matrix, we validated the utility of ACC tumor constructs by integrating fluorogenic MMP-sensitive peptide biosensors within the tumor constructs. Lastly, in our metastasis-on-a-chip device, NCI-H295R cells successfully engrafted in a downstream lung cell line-based construct, but invasion distance into the lung construct was decreased by MMP inhibition. These studies, which would not be possible using 2D cell cultures, demonstrated that NCI-H295R cells secreted active MMPs that are used for invasion in 3D. This work represents the first evidence of a 3D tumor constructs platform for ACC that can be deployed for future mechanistic studies as well as development of new targets for intervention and therapies.
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