PO.TB10.12 · 肿瘤生物学
在3D肿瘤支架中模拟三阴性乳腺癌的细胞外基质
Modeling the triple-negative breast cancer extracellular matrix in a 3d tumor scaffold
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
乳腺癌是女性癌症死亡的第二大原因，发病率每年增长0.5%。三阴性乳腺癌（TNBC）以激素受体（HR-）、孕激素和雌激素以及人表皮生长因子受体2（HER2）阴性为分类特征，是最具侵袭性的亚型，更常见于年轻的黑人和西班牙裔女性。与其他亚型相比，TNBC因治疗反应率仅为30%而难以治疗。新辅助化疗是TNBC的标准治疗。这归因于缺乏能够在TNBC高度异质性背景下实现多细胞相互作用的临床前模型。因此，迫切需要能够重现复杂细胞外基质（ECM）的3D TNBC肿瘤模型。大多数ECM-癌症研究聚焦于collagen I（COL1）；然而，COL1并非TNBC的预后标志物，且在TNBC中常未观察到升高。与相邻脂肪组织相比，评估原发性TNBC肿瘤中过表达的ECM显示ECM蛋白表达升高。为确定不同ECM对TNBC乳腺癌进展的影响，我们探究了TNBC细胞系中ECM蛋白表达（MFAP5、FN1和POSTN）的变化，并为体外系统制备了基质。通过qRT-PCR和RNA测序发现，在2D培养中，ECM-OE表达细胞中与上皮-间质转化和炎症相关的基因显著上调。随后，我们通过制备ECM基质包被的细胞培养板（COL1、FN1和POSTN）以及使用生物惰性鱼明胶甲基丙烯酰基（fgelMA）的3D支架，构建了TNBC ECM的3D模型。综上，这项工作确定了ECM蛋白——MFAP5、FN1和POSTN——通过上皮-间质转化促成TNBC增殖和存活。总体而言，我们的结果确立了三阴性肿瘤类型的细胞外特征，同时也证明所制备的3D ECM模型可用于推进TNBC潜在治疗方法的开发。
查看英文原文 English abstract
Breast Cancer is the second leading cause of cancer death in women and incidence rates are increasing 0.5% per year. Triple negative breast cancer (TNBC), which is classified by negative hormone receptors (HR-), progesterone and estrogen, and human epidermal growth factor receptor 2 (HER2) negative, is the most aggressive subtype and occurs more often in younger Black and Hispanic women. TNBC, compared to other subtypes, is difficult to treat due to the treatment response rate of 30%. Neoadjuvant chemotherapy is the standard treatment for TNBC. This is attributed to the lack of preclinical models that allow multi-cellular interactions in the highly heterogenous nature of TNBC. Due to this, there is a critical need for 3D TNBC tumor models that recapitulate the complex extracellular matrix (ECM). Most ECM-cancer studies focus on collagen I (COL1); however, COL1 is not a prognostic marker for TNBC and is often not observed to be elevated in TNBC. Evaluation of ECMs overexpressed in a primary TNBC tumor compared to adjacent adipose demonstrated elevated ECM protein expression. To determine the impact of distinct ECMs in TNBC breast cancer progression, we explored changes in ECM protein expression (MFAP5, FN1, and POSTN) in TNBC cell lines and fabricated matrix for in vitro systems. qRT-PCR and RNA sequencing, genes associated with epithelial-to-mesenchymal transition and inflammation were significantly upregulated in ECM-OE expressing cells in 2D culture. Next, we created 3D models of TNBC ECM through the fabrication of ECM matrix coated cell culture plates (COL1, FN1, and POSTN) and 3D scaffolds using bioinert fish gelatin methacrylol (fgelMA). Taken together, this work identifies ECM proteins - MFAP5, FN1, and POSTN - attribute to TNBC proliferation and survival through epithelial to mesenchymal transition. Overall, our results establish an extracellular signature for the triple-negative tumor type while also demonstrating fabricated 3D ECM models can be used to advancing the development of potential treatments for TNBC.
利益披露 Disclosure
K. Hebert, None..
T. Cheng, None..
S. Dietrich, None..
M. Hawes, None..
W. Monroe, None..
J. Belgodere, None.