Table_5_Establishment of Organoids From Human Epithelioid Sarcoma With the Air-Liquid Interface Organoid Cultures.xlsx

BackgroundAlthough biological resources are essential for basic and preclinical research in the oncological field, those of sarcoma are not sufficient for rapid development of the treatment. So far, some sarcoma cell lines have been established, however, the success rate was low and the established sarcoma types were frequently biased. Therefore, an efficient culture method is needed to determine the various types of sarcomas. Organoid culture is a 3-dimentional culture method that enables the recapitulation of the tumor microenvironment and the success rate reported is higher than the 2-dimentional culture. The purpose of this study was to report our newly established organoids from human epithelioid sarcoma using the air-liquid interface organoid culture method.
MethodsWe treated 2 patients with epithelioid sarcoma in our institute. The remaining sarcoma specimens after surgical resection were embedded in collagen type 1 gels according to the air-liquid interface organoid culture method. After serial passages, we xenografted the organoids to NOD-scid IL2Rgnull (NSG) mice. Using the developed tumors, we performed histological and genomic analyses to compare the similarities and differences with the original epithelioid sarcoma from the patient.
ResultsOrganoids from the epithelioid sarcoma could be serially cultured and maintained in collagen type 1 gels for more than 3 passages. Developed orthotopic tumor xenografts were detected in the NSG mice. After the process was repeated severally, the patient derived organoid lines from the epithelioid sarcoma were established. The established organoids showed loss of integrase interactor 1 expression with polymerase chain reaction and immunohistochemical analyses. The xenografted organoids of the epithelioid sarcoma had histologically similar phenotypes with the original tumor and genetically resembled it to some degree.
ConclusionsThe present study demonstrated 2 novel established organoid models of epithelioid sarcoma, and our organoid models could be used to investigate the molecular pathogenesis and develop a novel treatment.

【背景】尽管生物资源对于肿瘤学领域的基础研究与临床前研究至关重要，但肉瘤相关生物资源却难以满足治疗快速研发的需求。截至目前，虽已有部分肉瘤细胞系被成功构建，但其构建成功率较低，且所获肉瘤细胞系的类型常存在偏倚。因此，亟需开发高效的培养方法以适配各类肉瘤的研究需求。类器官（organoid）培养是一种三维（3D）培养技术，可重现肿瘤微环境，且已有研究显示其构建成功率高于二维（2D）培养。本研究旨在报告我们利用气液界面类器官培养方法，从人类上皮样肉瘤中成功构建的新型类器官。【方法】本研究纳入本院收治的2例上皮样肉瘤患者，将手术切除后剩余的肉瘤标本按照气液界面类器官培养方案，包埋于Ⅰ型胶原凝胶中。经过连续传代后，将类器官移植至NOD-scid IL2Rgnull（NSG）免疫缺陷小鼠体内构建异种移植瘤。利用形成的异种移植瘤，我们开展组织学与基因组学分析，以对比其与患者原发上皮样肉瘤的异同。【结果】本研究成功实现上皮样肉瘤类器官的连续传代培养，可在Ⅰ型胶原凝胶中维持传代超过3次。NSG小鼠体内可检测到原位异种移植瘤。经过多次重复该流程后，我们成功建立了上皮样肉瘤患者来源的类器官株系。通过聚合酶链反应（polymerase chain reaction, PCR）与免疫组化分析证实，所构建的类器官整合酶相互作用蛋白1（integrase interactor 1）表达缺失。上皮样肉瘤异种移植瘤的组织学表型与原发肿瘤高度相似，且在遗传学层面也与原发肿瘤具有一定程度的一致性。【结论】本研究成功建立2株新型上皮样肉瘤类器官模型，该类器官模型可用于探究上皮样肉瘤的分子发病机制，并助力新型治疗方案的开发。