Development of primary human pancreatic cancer organoids, matched stromal and immune cells and 3D tumor microenvironment models.
Development of primary human pancreatic cancer organoids, matched stromal and immune cells and 3D tumor microenvironment models.
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DOI：
10.1186/s12885-018-4238-4
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发表时间：
2018-03-27
期刊：
影响因子：
3.8
通讯作者：
James MA
中科院分区：
文献类型：
--
作者：
Tsai S;McOlash L;Palen K;Johnson B;Duris C;Yang Q;Dwinell MB;Hunt B;Evans DB;Gershan J;James MA
患者来源的肿瘤模型是临床前药物测试和生物标记物发现的新标准。然而，新兴的原发胰腺癌有机化合物技术尚未在研究中广泛应用，使用有机化合物与肿瘤间质和免疫细胞成分共培养的复杂器官模型尚未建立。在这项研究中，我们的目标是建立和表征胰腺癌类器官和多细胞类型的器官型共培养模型，以证明它们在胰腺癌研究中的适用性。我们使用类器官培养方法以及流式细胞学、细胞学、免疫荧光和免疫组织化学方法来建立和表征胰腺癌患者来源的类器官和肿瘤微环境的多细胞类型共培养模型。我们描述了来自切除、腹水和快速尸检来源的人胰腺癌有机类物质的培养和特征，以及来自这些来源的贴壁肿瘤细胞单培养和肿瘤相关成纤维细胞的来源。原代人类器官表现出肿瘤样的细胞形态、组织结构和极性，而细胞系球体形成均匀的、无腔形成的球体。重要的是，我们展示了构建复杂的器官模型的肿瘤，间质和免疫成分的肿瘤微环境。在这些模型中，可观察到肌成纤维样癌相关成纤维细胞的激活和肿瘤依赖性淋巴细胞的浸润。这些研究首次报道了代表胰腺肿瘤、间质和免疫成分的新型和疾病相关的3D体外模型，该模型使用代表肿瘤-微环境的原代有机化合物共培养。这些模型有望促进肿瘤-间质和肿瘤-免疫相互作用的研究，并可能在T细胞渗透的背景下评估免疫疗法，如检查点抑制剂。
Patient-derived tumor models are the new standard for pre-clinical drug testing and biomarker discovery. However, the emerging technology of primary pancreatic cancer organoids has not yet been broadly implemented in research, and complex organotypic models using organoids in co-culture with stromal and immune cellular components of the tumor have yet to be established. In this study, our objective was to develop and characterize pancreatic cancer organoids and multi-cell type organotypic co-culture models to demonstrate their applicability to the study of pancreatic cancer. We employed organoid culture methods and flow cytometric, cytologic, immunofluorescent and immunohistochemical methods to develop and characterize patient-derived pancreatic cancer organoids and multi-cell type organotypic co-culture models of the tumor microenvironment. We describe the culture and characterization of human pancreatic cancer organoids from resection, ascites and rapid autopsy sources and the derivation of adherent tumor cell monocultures and tumor-associated fibroblasts from these sources. Primary human organoids displayed tumor-like cellular morphology, tissue architecture and polarity in contrast to cell line spheroids, which formed homogenous, non-lumen forming spheres. Importantly, we demonstrate the construction of complex organotypic models of tumor, stromal and immune components of the tumor microenvironment. Activation of myofibroblast-like cancer associated fibroblasts and tumor-dependent lymphocyte infiltration were observed in these models. These studies provide the first report of novel and disease-relevant 3D in-vitro models representing pancreatic tumor, stromal and immune components using primary organoid co-cultures representative of the tumor-microenvironment. These models promise to facilitate the study of tumor-stroma and tumor-immune interaction and may be valuable for the assessment of immunotherapeutics such as checkpoint inhibitors in the context of T-cell infiltration.
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煤矿中的金丝雀：小鼠中患者衍生的肿瘤的生长预测胰腺导管腺癌手术切除后的临床复发。
影响因子：
3.7
作者：
Thomas RM;Truty MJ;Kim M;Kang Y;Zhang R;Chatterjee D;Katz MH;Fleming JB
通讯作者：
Fleming JB
影响因子：
7.2
作者：
Lunardi S;Lim SY;Muschel RJ;Brunner TB
通讯作者：
Brunner TB
DOI：
10.1073/pnas.95.2.499
发表时间：
1998-01-20
影响因子：
11.1
作者：
Sheng, SJ;Truong, B;Sager, R
通讯作者：
Sager, R
影响因子：
82.9
作者：
Huang, Ling;Holtzinger, Audrey;Muthuswamy, Senthil K.
通讯作者：
Muthuswamy, Senthil K.
影响因子：
11.2
作者：
James MA;Vikis HG;Tate E;Rymaszewski AL;You M
通讯作者：
You M