A CRISPR/Cas9-Engineered ARID1A-Deficient Human Gastric Cancer Organoid Model Reveals Essential and Nonessential Modes of Oncogenic Transformation.
A CRISPR/Cas9-Engineered ARID1A-Deficient Human Gastric Cancer Organoid Model Reveals Essential and Nonessential Modes of Oncogenic Transformation.
复制标题
CRISPR/Cas9工程的ARID1A缺陷型人胃癌器官模型揭示了致癌转化的必要和非必要模式。
DOI：
10.1158/2159-8290.cd-20-1109
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发表时间：
2021-06
期刊：
影响因子：
28.2
通讯作者：
Kuo CJ
中科院分区：
文献类型：
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作者：
Lo YH;Kolahi KS;Du Y;Chang CY;Krokhotin A;Nair A;Sobba WD;Karlsson K;Jones SJ;Longacre TA;Mah AT;Tercan B;Sockell A;Xu H;Seoane JA;Chen J;Shmulevich I;Weissman JS;Curtis C;Califano A;Fu H;Crabtree GR;Kuo CJ
ARID 1A突变是人类癌症中最常见的分子畸变之一。然而，由于缺乏正向遗传模型，人类细胞中ARID 1A突变的致癌后果仍然不清楚。在这里，CRISPR/Cas9介导的ARID 1A敲除在原代TP 53 −/−人胃类器官中诱导形态发育不良、致瘤性和粘液分化。遗传Wnt/β-连环蛋白激活挽救了粘液分化，但没有过度增殖，表明ARID 1A KO介导的转化的替代途径。ARID 1A突变诱导MSI和EBV亚型人胃癌特征性转录调控模块，包括FOXM 1相关有丝分裂基因和BIRC 5/生存素。聚合，高通量化合物筛选表明ARID 1A缺陷类器官对BIRC 5/生存素抑制的选择性脆弱性，在功能上暗示该途径是ARID 1A KO依赖性早期胃癌发生的重要介质。总的来说，我们定义了致癌ARID 1A突变下游的不同途径，非必需的Wnt抑制的粘液分化与必需的转录FOXM 1/BIRC 5刺激的增殖平行，说明了基于类器官的正向遗传癌症分析在人类细胞中的一般效用。
Mutations in ARID1A rank amongst the most common molecular aberrations in human cancer. However, oncogenic consequences of ARID1A mutation in human cells remain poorly defined due to lack of forward genetic models. Here, CRISPR/Cas9-mediated ARID1A knockout in primary TP53−/− human gastric organoids induced morphologic dysplasia, tumorigenicity and mucinous differentiation. Genetic Wnt/β-catenin activation rescued mucinous differentiation, but not hyperproliferation, suggesting alternative pathways of ARID1A KO-mediated transformation. ARID1A mutation induced transcriptional regulatory modules characteristic of MSI and EBV subtype human gastric cancer, including FOXM1-associated mitotic genes and BIRC5/survivin. Convergently, high-throughput compound screening indicated selective vulnerability of ARID1A-deficient organoids to inhibition of BIRC5/survivin, functionally implicating this pathway as an essential mediator of ARID1A KO-dependent early-stage gastric tumorigenesis. Overall, we define distinct pathways downstream of oncogenic ARID1A mutation, with non-essential Wnt-inhibited mucinous differentiation in parallel with essential transcriptional FOXM1/BIRC5-stimulated proliferation, illustrating the general utility of organoid-based forward genetic cancer analysis in human cells.