Detection of lineage-reprogramming efficiency of tumor cells in a 3D-printed liver-on-a-chip model.
Detection of lineage-reprogramming efficiency of tumor cells in a 3D-printed liver-on-a-chip model.
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DOI：
10.7150/thno.86921
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发表时间：
2023
期刊：
影响因子：
12.4
通讯作者：
Atala A
中科院分区：
文献类型：
--
作者：
Lu Z;Miao X;Song Q;Ding H;Rajan SAP;Skardal A;Votanopoulos KI;Dai K;Zhao W;Lu B;Atala A
背景：肝脏转移伴随着肝功能的丧失是三阴性乳腺癌患者（TNBC）最常见的并发症之一。谱系重编程作为一种直接诱导功能性细胞类型的技术，而无需通过中间多能阶段，在改变细胞命运并克服原代细胞的局限性方面有望。但是，大多数重编程技术来自人类成纤维细胞，以及是否可以将癌细胞逆转为肝细胞仍然难以捉摸。
方法：本文，我们通过从两个慢病毒载体中表达六个转录因子（HNF4A，FOXA2，FOXA3，Prox1和HNF1）来简化重编程试剂的制备，每个都表达了三个因素。然后将病毒转导至MDA-MB-231细胞中，以产生人类诱导的肝细胞样细胞（HIHEPS），并使用单细胞测序来分析重新编程后的细胞命运。此外，我们通过将带有肝细胞外囊泡（Gelma-ev）生物学的明胶甲基丙烯酰基水凝胶生物构建，构建了一个肝脏（LOC）模型（LOC）模型。肝微环境体外。
结果：HNF4A，FOXA2，FOXA3，ATF5，PROX1和HNF1A基因的组合可以将MDA-MB-231肿瘤细胞重新编程为人类诱导的肝细胞（HIHEPS），从而限制了这些细胞的转移。单细胞测序分析表明，在谱系重编程后激活肝脏特异性基因时，致癌基因受到显着抑制。最后，构造的LOC模型表明，可以观察到重编程细胞的肝表型，并且在肝微环境下可以抑制嵌入的癌细胞的转移。
结论：我们的发现表明，重编程可能是产生肝细胞和治疗TNBC肝转移的有前途的方法。 LOC模型可以牢固3D肝微环境并评估重编程的TNBC细胞的行为。
Background: The liver metastasis accompanied with the loss of liver function is one of the most common complications in patients with triple-negative breast cancers (TNBC). Lineage reprogramming, as a technique direct inducing the functional cell types from one lineage to another lineage without passing through an intermediate pluripotent stage, is promising in changing cell fates and overcoming the limitations of primary cells. However, most reprogramming techniques are derived from human fibroblasts, and whether cancer cells can be reversed into hepatocytes remains elusive. Methods: Herein, we simplify preparation of reprogramming reagents by expressing six transcriptional factors (HNF4A, FOXA2, FOXA3, ATF5, PROX1, and HNF1) from two lentiviral vectors, each expressing three factors. Then the virus was transduced into MDA-MB-231 cells to generated human induced hepatocyte-like cells (hiHeps) and single-cell sequencing was used to analyze the fate for the cells after reprogramming. Furthermore, we constructed a Liver-on-a-chip (LOC) model by bioprinting the Gelatin Methacryloyl hydrogel loaded with hepatocyte extracellular vesicles (GelMA-EV) bioink onto the microfluidic chip to assess the metastasis behavior of the reprogrammed TNBC cells under the 3D liver microenvironment in vitro. Results: The combination of the genes HNF4A, FOXA2, FOXA3, ATF5, PROX1 and HNF1A could reprogram MDA-MB-231 tumor cells into human-induced hepatocytes (hiHeps), limiting metastasis of these cells. Single-cell sequencing analysis showed that the oncogenes were significantly inhibited while the liver-specific genes were activated after lineage reprogramming. Finally, the constructed LOC model showed that the hepatic phenotypes of the reprogrammed cells could be observed, and the metastasis of embedded cancer cells could be inhibited under the liver microenvironment. Conclusion: Our findings demonstrate that reprogramming could be a promising method to produce hepatocytes and treat TNBC liver metastasis. And the LOC model could intimate the 3D liver microenvironment and assess the behavior of the reprogrammed TNBC cells.
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影响因子：
37.3
作者：
Liu K;Cui JJ;Zhan Y;Ouyang QY;Lu QS;Yang DH;Li XP;Yin JY
通讯作者：
Yin JY
影响因子：
9
作者：
Lu, Zuyan;Jiang, Xia;Kang, Y. James
通讯作者：
Kang, Y. James
影响因子：
13.5
作者：
Oertel, M;Rosencrantz, R;Shafritz, DA
通讯作者：
Shafritz, DA
影响因子：
16.6
作者：
Montaudon, Elodie;Nikitorowicz-Buniak, Joanna;Marangoni, Elisabetta
通讯作者：
Marangoni, Elisabetta
影响因子：
50.3
作者：
Ishay-Ronen, Dana;Diepenbruck, Maren;Christofori, Gerhard
通讯作者：
Christofori, Gerhard