Organoid-based ex vivo reconstitution of Kras-driven pancreatic ductal carcinogenesis
Organoid-based ex vivo reconstitution of Kras-driven pancreatic ductal carcinogenesis
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DOI：
10.1093/carcin/bgz122
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发表时间：
2020-04-01
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：
Matsuura, Tetsuya;Maru, Yoshiaki;Hippo, Yoshitaka
类器官培养技术最近已被应用于模拟几种器官的癌变过程。为了进一步探索其潜力并获得肿瘤发生的新见解，我们在这里研究了胰腺导管腺癌（PDA）是否可以通过正常类器官的基因工程在免疫功能低下的裸鼠中作为皮下肿瘤产生。正如预期的那样，体外急性诱导KrasG12D偶尔会导致与早期病变相适应的微小结节的发展，称为胰腺上皮内瘤变（PanIN）。krasg12d表达细胞在皮下接种后富集，但在培养过程中却明显减少，提示其优势可能与周围环境有关。生长因子的缺失或Trp53的同时缺失导致其强烈富集，分别不可避免地导致PanIN或大级别腺癌的发展，这与同一基因型的小鼠体内研究一致。PanIN的进展也可以通过随后敲低常见肿瘤抑制因子来重现，而Tgfbr2缺失的影响仅部分重现，这说明了胰腺生态位对肿瘤发生的基因型依赖性需求。有趣的是，对肿瘤衍生类器官的分析显示，krasg12d表达细胞自发缺失野生型Kras，在裸鼠中被积极选择，并表现出与衰老相关的突变特征，反映了人类PDA的发病机制，并且在同基因小鼠中球体形成潜力和原位致瘤性显着增强。这些观察结果强调了裸鼠皮下促进PDA发展的相关性，尽管其异位性。总之，胰腺癌的发生可以用类器官来概括，这可能是一种新的疾病模型。
The organoid culture technique has been recently applied to modeling carcinogenesis in several organs. To further explore its potential and gain novel insights into tumorigenesis, we here investigated whether pancreatic ductal adenocarcinoma (PDA) could be generated as subcutaneous tumors in immunocompromised nude mice, by genetic engineering of normal organoids. As expected, acute induction of KrasG12D in vitro occasionally led to development of tiny nodules compatible with early lesions known as pancreatic intraepithelial neoplasia (PanIN). KrasG12D-expressing cells were enriched after inoculation in the subcutis, yet proved rather declined during culture, suggesting that its advantage might depend on surrounding environments. Depletion of growth factors or concurrent Trp53 deletion resulted in its robust enrichment, invariably leading to development of PanIN or large high-grade adenocarcinoma, respectively, consistent with in vivo mouse studies for the same genotype. Progression from PanIN was also recapitulated by subsequent knockdown of common tumor suppressors, whereas the impact of Tgfbr2 deletion was only partially recapitulated, illustrating genotype-dependent requirement of the pancreatic niche for tumorigenesis. Intriguingly, analysis of tumor-derived organoids revealed that KrasG12D-expressing cells with spontaneous deletion of wild-type Kras were positively selected and exhibited an aging-related mutation signature in nude mice, mirroring the pathogenesis of human PDA, and that the sphere-forming potential and orthotopic tumorigenicity in syngenic mice were significantly augmented. These observations highlighted the relevance of the subcutis of nude mice in promoting PDA development despite its ectopic nature. Taken together, pancreatic carcinogenesis could be considerably recapitulated with organoids, which would probably serve as a novel disease model.