Background & Aims

Cancer associated fibroblasts (CAFs) play a key role in cancer process, but the research progress is hampered by the paucity of preclinical models essential for mechanistic dissection of cancer cell-CAF interactions. Here, we aim to establish a 3D organotypic co-cultures of primary liver tumor-derived organoids with CAFs, and to understand their interactions and the response to treatment.

Methods

Liver tumor organoids and CAFs were cultured from murine and human primary liver tumors. 3D co-culture models of tumor organoids with CAFs and trans-well culture systems were established in vitro. A xenograft model was used to interrogate the cell-cell interactions in vivo. Gene expression analysis of CAF markers in our hepatocellular carcinoma (HCC) cohort and an online liver cancer database indicated the clinical relevance of CAFs.

Results

To functionally investigate the interactions of liver cancer cells with CAFs, we have successfully established murine and human 3D co-culture models of liver tumor organoids with CAFs. CAFs promoted tumor organoid growth in co-culture with direct cell-cell contact and in trans-well system via paracrine signaling. Vice versa, cancer cells secret paracrine factors regulating CAF physiology. Co-transplantation of CAFs with liver tumor organoids of mouse or human origin promoted tumor growth in xenograft models. Moreover, tumor organoids conferred resistance to clinically used anti-cancer drugs including Sorafenib, Regorafenib and Fluorouracil (5-FU) in the presence of CAFs, or the conditioned medium of CAFs.

Conclusions

We have successfully established murine and human 3D co-culture models and have revealed robust effects of CAFs in liver cancer nurturing and treatment resistance.

中文翻译：

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癌症相关成纤维细胞为肝癌类器官提供了一个基质生态位，赋予了营养效应和治疗抗性。

背景与目标

癌症相关成纤维细胞 (CAF) 在癌症过程中起着关键作用，但由于缺乏对癌细胞-CAF 相互作用的机械解剖必不可少的临床前模型，研究进展受到阻碍。在这里，我们的目标是建立原发性肝肿瘤衍生类器官与 CAF 的 3D 器官型共培养，并了解它们的相互作用和对治疗的反应。

方法

从小鼠和人类原发性肝脏肿瘤中培养出肝肿瘤类器官和 CAF。在体外建立了肿瘤类器官与 CAF 和跨孔培养系统的 3D 共培养模型。异种移植模型用于研究体内细胞间相互作用。我们的肝细胞癌 (HCC) 队列和在线肝癌数据库中 CAF 标志物的基因表达分析表明了 CAF 的临床相关性。

结果

为了从功能上研究肝癌细胞与 CAFs 的相互作用，我们成功地建立了肝肿瘤类器官与 CAFs 的小鼠和人类 3D 共培养模型。CAFs 在与细胞间直接接触的共培养和跨孔系统中通过旁分泌信号促进肿瘤类器官生长。反之亦然，癌细胞分泌调节 CAF 生理的旁分泌因子。CAFs 与小鼠或人类来源的肝肿瘤类器官的共同移植促进了异种移植模型中的肿瘤生长。此外，在 CAFs 或 CAFs 条件培养基存在下，肿瘤类器官对临床使用的抗癌药物包括索拉非尼、瑞戈非尼和氟尿嘧啶 (5-FU) 具有抗性。

结论

我们已经成功建立了小鼠和人类 3D 共培养模型，并揭示了 CAFs 在肝癌培育和治疗抵抗中的强大作用。