Modeling Breast Cancer in Human Breast Tissue using a Microphysiological System
Modeling Breast Cancer in Human Breast Tissue using a Microphysiological System
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DOI：
10.3791/62009
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发表时间：
2021-04-01
影响因子：
1.2
通讯作者：
Lau, Frank H.
中科院分区：
文献类型：
--
作者：
Brown, Loren M.;Hebert, Katherine L.;Lau, Frank H.
乳腺癌（BC）仍然是女性死亡的主要原因。尽管每年在 BC 研究上的投资超过 7 亿美元，但 97% 的 BC 候选药物未能通过临床试验。因此，需要新的模型来提高我们对这种疾病的认识。 NIH 微生理系统 (MPS) 项目的开发是为了改善基础科学发现和有前景的新治疗策略的临床转化。在这里，我们提出了一种生成乳腺癌 MPS 的方法 (BC-MPS)。该模型采用了先前描述的通过将 WAT 夹在脂肪干细胞片 (ASC) 之间来培养原代人类白色脂肪组织 (WAT) 的方法。我们的 BC-MPS 的新颖之处包括将 BC 细胞接种到含有天然细胞外基质、成熟脂肪细胞、常驻成纤维细胞和免疫细胞的非患病人类乳腺组织 (HBT) 中；并将BC-HBT混合物夹在HBT衍生的ASC片材之间。所得 BC-MPS 在离体培养物中稳定至少 14 天。该模型系统包含影响 BC 的微环境的多种元素，包括脂肪细胞、基质细胞、免疫细胞和细胞外基质。因此，BC-MPS 可用于研究 BC 与其微环境之间的相互作用。我们通过研究已知影响癌症进展和转移的两种 BC 行为来证明 BC-MPS 的优势：1）BC 运动性和 2）BC-HBT 代谢串扰。虽然 BC 运动性先前已通过活体成像得到证实，但 BC-MPS 允许使用荧光显微镜在几天内进行高分辨率延时成像。此外，虽然之前使用 BC 细胞和分化为未成熟脂肪细胞的小鼠前脂肪细胞证明了代谢串扰，但我们的 BC-MPS 模型是第一个在体外证明原代人乳腺脂肪细胞和 BC 细胞之间这种串扰的系统。
Breast cancer (BC) remains a leading cause of death for women. Despite more than $700 million invested in BC research annually, 97% of candidate BC drugs fail clinical trials. Therefore, new models are needed to improve our understanding of the disease. The NIH Microphysiological Systems (MPS) program was developed to improve the clinical translation of basic science discoveries and promising new therapeutic strategies. Here we present a method for generating MPS for breast cancers (BC-MPS). This model adapts a previously described approach of culturing primary human white adipose tissue (WAT) by sandwiching WAT between adipose-derived stem cell sheets (ASC)s. Novel aspects of our BC-MPS include seeding BC cells into non-diseased human breast tissue (HBT) containing native extracellular matrix, mature adipocytes, resident fibroblasts, and immune cells; and sandwiching the BC-HBT admixture between HBT-derived ASC sheets. The resulting BC-MPS is stable in culture ex vivo for at least 14 days. This model system contains multiple elements of the microenvironment that influence BC including adipocytes, stromal cells, immune cells, and the extracellular matrix. Thus BC-MPS can be used to study the interactions between BC and its microenvironment.We demonstrate the advantages of our BC-MPS by studying two BC behaviors known to influence cancer progression and metastasis: 1) BC motility and 2) BC-HBT metabolic crosstalk. While BC motility has previously been demonstrated using intravital imaging, BC-MPS allows for high-resolution time-lapse imaging using fluorescence microscopy over several days. Furthermore, while metabolic crosstalk was previously demonstrated using BC cells and murine pre-adipocytes differentiated into immature adipocytes, our BC-MPS model is the first system to demonstrate this crosstalk between primary human mammary adipocytes and BC cells in vitro.