Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell- and patient-derived tumor organoids
Ductal pancreatic cancer modeling and drug screening using human pluripotent stem cell- and patient-derived tumor organoids
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DOI：
10.1038/nm.3973
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发表时间：
2015-11-01
期刊：
影响因子：
82.9
通讯作者：
Muthuswamy, Senthil K.
中科院分区：
文献类型：
--
作者：
Huang, Ling;Holtzinger, Audrey;Muthuswamy, Senthil K.
外分泌胰腺发育和原发性人类胰腺腺癌（PDAC）的体外模型很少。我们建立了三维培养条件，以诱导人类多能干细胞向外分泌祖细胞类器官分化，这些类器官在体外培养和体内形成导管和腺泡结构。在祖细胞类器官中表达突变的KRAS或TP53会在体外培养和体内诱导出突变特异性表型。TP53（R175H）的表达会诱导胞质内SOX9定位。在携带TP53突变的患者肿瘤中，SOX9位于细胞质中且与死亡率相关。我们还确定了从新鲜切除的PDAC中克隆生成肿瘤类器官的培养条件。肿瘤类器官保持了原发性肿瘤的分化状态、组织结构和表型异质性，并保留了患者特异性的生理变化，包括缺氧、耗氧量、表观遗传标记以及对组蛋白甲基转移酶EZH2抑制敏感性的差异。因此，胰腺祖细胞类器官和肿瘤类器官可用于模拟PDAC并进行药物筛选，以确定精准治疗策略。
There are few in vitro models of exocrine pancreas development and primary human pancreatic adenocarcinoma (PDAC). We establish three-dimensional culture conditions to induce the differentiation of human pluripotent stem cells into exocrine progenitor organoids that form ductal and acinar structures in culture and in vivo. Expression of mutant KRAS or TP53 in progenitor organoids induces mutation-specific phenotypes in culture and in vivo. Expression of TP53(R175H) induces cytosolic SOX9 localization. In patient tumors bearing TP53 mutations, SOX9 was cytoplasmic and associated with mortality. We also define culture conditions for clonal generation of tumor organoids from freshly resected PDAC. Tumor organoids maintain the differentiation status, histoarchitecture and phenotypic heterogeneity of the primary tumor and retain patient-specific physiological changes, including hypoxia, oxygen consumption, epigenetic marks and differences in sensitivity to inhibition of the histone methyltransferase EZH2. Thus, pancreatic progenitor organoids and tumor organoids can be used to model PDAC and fat drug screening to identify precision therapy strategies.