Kras-driven heterotopic tumor development from hepatobiliary organoids
Kras-driven heterotopic tumor development from hepatobiliary organoids
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DOI：
10.1093/carcin/bgz024
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发表时间：
2019-09-01
期刊：
影响因子：
4.7
通讯作者：
Hippo, Yoshitaka
中科院分区：
文献类型：
--
作者：
Ochiai, Masako;Yoshihara, Yasunori;Hippo, Yoshitaka
胆道癌是传统治疗难以治愈的，需要开发新的治疗方法。然而，只有有限数量的基因工程小鼠模型已经创建，部分原因是需要耗时的工作。此外，肝脏特异性基因操作主要导致另一种类型的肝癌肝细胞癌的并发发展，胆囊限制性基因靶向仍然不可行。因此，建立癌症类型特异性疾病模型仍然是一个技术挑战。为了解决这个问题，我们采取了另一种基于细胞的方法来快速诱导离体肿瘤发生。具体而言，用慢病毒载体转导来自肝脏和胆囊的鼠原发性类器官，以重建胆道癌中常见的遗传改变，然后在免疫缺陷小鼠中接种。虽然任何单一的遗传改变不会诱发肿瘤，但突变型Kras和主要肿瘤抑制因子的抑制在2个月内协同促进肿瘤的发展。诱导的病变在正常、异型增生和乳头状病变到腺癌之间各不相同，即使在异位情况下也重复了多步骤的肿瘤发生。我们进一步证明了肝内胆管细胞癌中的两种推定癌基因，突变型Pik 3ca和FGFR 2-AHCYL 1融合，是肝源性类器官的相当温和的驱动因素，可能需要额外的突变或肝小生境来稳健地诱导完全成熟的肿瘤。因此，我们表明，癌细胞可以很容易地从胆道中的原代细胞产生，至少在遗传因素起主导作用的情况下。总的来说，这项研究可能有助于获得胆管癌发生机制的见解，并为药物发现提供有价值的资源。
Cancers arising from the biliary tract are refractory to conventional therapies, requiring the development of novel therapeutics. However, only a limited number of genetically engineered mouse models have been created, partly because of time-consuming work required. Besides, liver-specific gene manipulation mostly resulted in concurrent development of hepatocellular carcinoma, another type of liver cancer, and gallbladder-restricted gene targeting is still not feasible. Consequently, establishment of cancer type-specific disease modeling remains a technical challenge. To address this issue, we took an alternative cell-based approach to quickly induce tumorigenesis ex vivo. Specifically, murine primary organoids from liver and gallbladder were transduced with lentiviral vectors to reconstitute genetic alterations common in biliary tract cancers, followed by inoculation in immunodeficient mice. Although any single genetic alteration did not induce tumors, mutant Kras and repression of major tumor suppressors cooperated for tumor development within 2 months. Induced lesions varied among normal, dysplastic and papillary lesions to adenocarcinoma, recapitulating multistep tumorigenesis even in a heterotopic situation. We further demonstrated that two putative oncogenes in intrahepatic cholangiocellular carcinoma, mutant Pik3ca and FGFR2-AHCYL1 fusion, were rather modest drivers for liver-derived organoids, probably requiring additional mutations or hepatic niche to robustly induce full-blown tumors. Thus, we showed that cancer cells could be readily generated from primary cells in the biliary tract, at least in cases where genetic factors play dominant roles. Collectively, this study will likely contribute to gaining mechanistic insights into biliary carcinogenesis and providing valuable resources for drug discovery.