Zhou Ruoji, Xu An, Tu Jian, Liu Mo, Gingold Julian A, Zhao Ruiying, Lee Dung-Fang
Department of Integrative Biology and Pharmacology, McGovern Medical School, The University of Texas Health Science Center at Houston; Graduate School of Biomedical Sciences, The University of Texas MD Anderson Cancer Center UTHealth.
Department of Integrative Biology and Pharmacology, McGovern Medical School, The University of Texas Health Science Center at Houston.
J Vis Exp. 2018 Jun 13(136):57664. doi: 10.3791/57664.
Li-Fraumeni syndrome (LFS) is an autosomal dominant hereditary cancer disorder. Patients with LFS are predisposed to a various type of tumors, including osteosarcoma--one of the most frequent primary non-hematologic malignancies in the childhood and adolescence. Therefore, LFS provides an ideal model to study this malignancy. Taking advantage of iPSC methodologies, LFS-associated osteosarcoma can be successfully modeled by differentiating LFS patient iPSCs to mesenchymal stem cells (MSCs), and then to osteoblasts--the cells of origin for osteosarcomas. These LFS osteoblasts recapitulate oncogenic properties of osteosarcoma, providing an attractive model system for delineating the pathogenesis of osteosarcoma. This manuscript demonstrates a protocol for the generation of iPSCs from LFS patient fibroblasts, differentiation of iPSCs to MSCs, differentiation of MSCs to osteoblasts, and in vivo tumorigenesis using LFS osteoblasts. This iPSC disease model can be extended to identify potential biomarkers or therapeutic targets for LFS-associated osteosarcoma.
李-弗劳梅尼综合征（LFS）是一种常染色体显性遗传性癌症疾病。LFS患者易患多种类型的肿瘤，包括骨肉瘤——儿童和青少年时期最常见的原发性非血液系统恶性肿瘤之一。因此，LFS为研究这种恶性肿瘤提供了一个理想模型。利用诱导多能干细胞（iPSC）技术，通过将LFS患者的iPSC分化为间充质干细胞（MSC），然后再分化为成骨细胞——骨肉瘤的起源细胞，可以成功构建与LFS相关的骨肉瘤模型。这些LFS成骨细胞概括了骨肉瘤的致癌特性，为描绘骨肉瘤的发病机制提供了一个有吸引力的模型系统。本手稿展示了从LFS患者成纤维细胞生成iPSC、将iPSC分化为MSC、将MSC分化为成骨细胞以及使用LFS成骨细胞进行体内肿瘤发生的方案。这种iPSC疾病模型可以扩展用于识别与LFS相关的骨肉瘤的潜在生物标志物或治疗靶点。
