In vivo genome editing and organoid transplantation models of colorectal cancer and metastasis.
In vivo genome editing and organoid transplantation models of colorectal cancer and metastasis.
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DOI：
10.1038/nbt.3836
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发表时间：
2017-06
影响因子：
46.9
通讯作者：
Yilmaz ÖH
中科院分区：
文献类型：
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作者：
Roper J;Tammela T;Cetinbas NM;Akkad A;Roghanian A;Rickelt S;Almeqdadi M;Wu K;Oberli MA;Sánchez-Rivera FJ;Park YK;Liang X;Eng G;Taylor MS;Azimi R;Kedrin D;Neupane R;Beyaz S;Sicinska ET;Suarez Y;Yoo J;Chen L;Zukerberg L;Katajisto P;Deshpande V;Bass AJ;Tsichlis PN;Lees J;Langer R;Hynes RO;Chen J;Bhutkar A;Jacks T;Yilmaz ÖH
由于需要产生和杂交种系突变小鼠，对与肿瘤发生有关的基因功能的体内研究受到限制。在这里，我们描述了建立结直肠癌（CRC）和转移模型的方法，这些方法依赖于没有癌症诱发突变的小鼠的原位基因编辑和原位类器官移植。结肠上皮细胞中基于 CRISPR-Cas9 的 Apc 和 Trp53 肿瘤抑制基因编辑以及 Apc 编辑的结肠类器官的原位移植可诱导原地肿瘤形成。 ApcΔ/ Δ;KrasG12D/+;Trp53Δ/ Δ (AKP) 小鼠结肠类器官和人 CRC 类器官植入远端结肠并转移至肝脏。最后，我们应用原位移植模型来表征 Lgr5+ 干细胞的克隆动力学，并证明癌基因在已建立的结肠腺瘤中的连续激活。这些实验系统能够快速体内表征癌症相关基因，并再现肿瘤进展和转移的整个范围。
In vivo interrogation of the function of genes implicated in tumorigenesis is limited by the need to generate and cross germline mutant mice. Here we describe approaches to model colorectal cancer (CRC) and metastasis that rely on in situ gene editing and orthotopic organoid transplantation in mice without cancer predisposing mutations. Autochthonous tumor formation is induced by CRISPR–Cas9-based editing of the Apc and Trp53 tumor suppressor genes in colon epithelial cells and by orthotopic transplantation of Apc-edited colon organoids. ApcΔ/ Δ;KrasG12D/+;Trp53Δ/ Δ (AKP) mouse colon organoids and human CRC organoids engraft in the distal colon and metastasize to the liver. Finally, we apply the orthotopic transplantation model to characterize the clonal dynamics of Lgr5+ stem cells and demonstrate sequential activation of an oncogene in established colon adenomas. These experimental systems enable rapid in vivo characterization of cancer-associated genes and reproduce the entire spectrum of tumor progression and metastasis.