Deligne Clémence, Tourbez Arthur, Bénard Flavie, Meyer Swann, Curt Alexis, Gianesello Matteo, Hamadou Maud, Clavier Léna, Coquet Claire, Bocquet Charlotte, Tomine Julia, Diot Thomas, Paraqindes Hermes, Marcel Virginie, Berthelot Clément, Engel Justine, Rochet Isabelle, Barritault Marc, Savary Clara, Gadot Nicolas, Attignon Valéry, Carrere Marjorie, Billaud Marc, Dutour Aurélie, Cordier-Bussat Martine, Beuriat Pierre-Aurélien, Szathmari Alexandru, Di Rocco Federico, Blay Jean-Yves, Tiberi Luca, Vasiljevic Alexandre, Meyronet David, Castets Marie, Leblond Pierre, Broutier Laura
Childhood Cancer & Cell Death Team (C3 Team), Consortium South-ROCK, LabEx DEVweCAN, Institut Convergence Plascan, Centre Léon Bérard, Centre de Recherche en Cancérologie de Lyon (CRCL), Université Claude Bernard Lyon 1, INSERM 1052, CNRS 5286, 69008 Lyon, France.
Armenise-Harvard Laboratory of Brain Disorders and Cancer, CIBIO, Trento, Italy.
Neuro Oncol. 2025 Jun 21;27(5):1325-1340. doi: 10.1093/neuonc/noaf007.
Brain tumors are the deadliest solid tumors in children and adolescents. Most of these tumors are glial in origin and exhibit strong heterogeneity, hampering the development of effective therapeutic strategies. In the past decades, patient-derived tumor organoids (PDT-O) have emerged as powerful tools for modeling tumoral cell diversity and dynamics, and they could then help define new therapeutic options for pediatric brain tumors.
Through an integrative approach based on our expertise and a careful review of the literature about glioblastoma 3D primary cultures, we set up a standardized methodological pipeline for the establishment, characterization, and biobanking of PDT-O through direct 3D in vitro culture of the deadliest pediatric glial brain tumors. To assess PDT-O fidelity and validate their preclinical relevance, we performed comprehensive histological, molecular, and drug-response analyses.
Our methodological pipeline allowed the rapid and efficient generation of PDT-O recapitulating their parental tumor features, including intratumoral heterogeneity, even after several passages and cryopreservation/revival as 3D cultures. Moreover, we successfully performed preclinical test responses on these PDT-O to standard-of-care therapies and new therapeutic options. Finally, we identified ONC201 as a selective drug for pediatric glial tumor types not restricted to H3K27-altered glial tumors, as well as combination strategies to increase the therapeutic response to ONC201.
Hence, we describe a fast and robust process to biobank PDT-O for pediatric glial brain tumors. These PDT-O models have the potential for patient-specific modeling even after long-term expansion in vitro, and we established the proof-of-concept of their usefulness to support powerful preclinical studies.
脑肿瘤是儿童和青少年中最致命的实体瘤。这些肿瘤大多起源于神经胶质细胞，具有很强的异质性，阻碍了有效治疗策略的发展。在过去几十年中，患者来源的肿瘤类器官（PDT-O）已成为模拟肿瘤细胞多样性和动态变化的有力工具，进而有助于为小儿脑肿瘤确定新的治疗选择。
通过基于我们专业知识的综合方法，并仔细回顾有关胶质母细胞瘤3D原代培养的文献，我们建立了一个标准化的方法流程，用于通过对最致命的小儿神经胶质脑肿瘤进行直接3D体外培养来建立、表征和生物样本保存PDT-O。为了评估PDT-O的保真度并验证其临床前相关性，我们进行了全面的组织学、分子和药物反应分析。
我们的方法流程能够快速高效地生成PDT-O，即使经过多次传代以及作为3D培养物进行冷冻保存/复苏后，仍能重现其亲本肿瘤的特征，包括肿瘤内异质性。此外，我们成功地对这些PDT-O进行了针对标准治疗方法和新治疗选择的临床前测试反应。最后，我们确定ONC201是一种针对小儿神经胶质肿瘤类型的选择性药物，不限于H3K27改变的神经胶质肿瘤，以及提高对ONC201治疗反应的联合策略。
因此，我们描述了一种快速且稳健的流程，用于为小儿神经胶质脑肿瘤生物样本保存PDT-O。即使在体外长期扩增后，这些PDT-O模型仍具有针对患者特异性建模的潜力，并且我们建立了其有用性的概念验证，以支持强大的临床前研究。
