器官型微流控乳腺癌模型揭示了饥饿诱导的时空代谢适应性。
Organotypic microfluidic breast cancer model reveals starvation-induced spatial-temporal metabolic adaptations.
机构信息
Morgridge Institute for Research, 330 N Orchard street, Madison, WI, USA; Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA; The University of Wisconsin Carbone Cancer Center, University of Wisconsin, Madison, WI, USA.
Department of Biomedical Engineering, University of Wisconsin, Madison, WI, USA.
出版信息
EBioMedicine. 2018 Nov;37:144-157. doi: 10.1016/j.ebiom.2018.10.046. Epub 2018 Oct 26.
BACKGROUND
Ductal carcinoma in situ (DCIS) is the earliest stage of breast cancer. During DCIS, tumor cells remain inside the mammary duct, growing under a microenvironment characterized by hypoxia, nutrient starvation, and waste product accumulation; this harsh microenvironment promotes genomic instability and eventually cell invasion. However, there is a lack of biomarkers to predict what patients will transition to a more invasive tumor or how DCIS cells manage to survive in this harsh microenvironment.
METHODS
In this work, we have developed a microfluidic model that recapitulates the DCIS microenvironment. In the microdevice, a DCIS model cell line was grown inside a luminal mammary duct model, embedded in a 3D hydrogel with mammary fibroblasts. Cell behavior was monitored by confocal microscopy and optical metabolic imaging. Additionally, metabolite profile was studied by NMR whereas gene expression was analyzed by RT-qPCR.
FINDINGS
DCIS cell metabolism led to hypoxia and nutrient starvation; revealing an altered metabolism focused on glycolysis and other hypoxia-associated pathways. In response to this starvation and hypoxia, DCIS cells modified the expression of multiple genes, and a gradient of different metabolic phenotypes was observed across the mammary duct model. These genetic changes observed in the model were in good agreement with patient genomic profiles; identifying multiple compounds targeting the affected pathways. In this context, the hypoxia-activated prodrug tirapazamine selectively destroyed hypoxic DCIS cells.
INTERPRETATION
The results showed the capacity of the microfluidic model to mimic the DCIS structure, identifying multiple cellular adaptations to endure the hypoxia and nutrient starvation generated within the mammary duct. These findings may suggest new potential therapeutic directions to treat DCIS. In summary, given the lack of in vitro models to study DCIS, this microfluidic device holds great potential to find new DCIS predictors and therapies and translate them to the clinic.
背景
导管原位癌（DCIS）是乳腺癌的最早阶段。在 DCIS 中，肿瘤细胞仍在乳腺导管内生长，在缺氧、营养饥饿和废物积累的微环境下生长；这种恶劣的微环境促进了基因组不稳定性，最终导致细胞侵袭。然而，目前缺乏预测哪些患者会发展为侵袭性更强的肿瘤的生物标志物，也缺乏了解 DCIS 细胞如何在这种恶劣的微环境中存活的生物标志物。
方法
在这项工作中，我们开发了一种微流控模型，该模型再现了 DCIS 微环境。在微器件中，将 DCIS 模型细胞系种植在腔道乳腺导管模型内，嵌入在含有乳腺成纤维细胞的 3D 水凝胶中。通过共聚焦显微镜和光学代谢成像监测细胞行为。此外，通过 NMR 研究代谢物谱，通过 RT-qPCR 分析基因表达。
结果
DCIS 细胞代谢导致缺氧和营养饥饿；揭示了一种以糖酵解和其他与缺氧相关的途径为重点的改变的代谢。为了应对这种饥饿和缺氧，DCIS 细胞改变了多个基因的表达，并在整个乳腺导管模型中观察到不同的代谢表型梯度。模型中观察到的这些遗传变化与患者的基因组图谱非常吻合；确定了针对受影响途径的多种化合物。在这种情况下，缺氧激活的前药替拉扎胺选择性地破坏了缺氧的 DCIS 细胞。
解释
结果表明，该微流控模型能够模拟 DCIS 结构，确定了多种细胞适应机制，以耐受乳腺导管内产生的缺氧和营养饥饿。这些发现可能为治疗 DCIS 提供新的潜在治疗方向。总之，鉴于缺乏用于研究 DCIS 的体外模型，这种微流控装置具有很大的潜力来寻找新的 DCIS 预测因子和治疗方法，并将其转化为临床应用。
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