肿瘤细胞治疗研究
英文原题：Patient-Derived Organoid Modeling of Glypican-3 CAR-T Responses in Hepatocellular Carcinoma.
Patient-Derived Organoid Modeling of Glypican-3 CAR-T Responses in Hepatocellular Carcinoma.
这些发现共同提供了概念验证证据，支持使用肝细胞癌 PDO 作为患者来源平台，用于模拟 GPC3 靶向 CAR-T 细胞活性的选定决定因素，并探索提高治疗疗效的联合策略。
靶向聚糖蛋白-3（GPC3）的CAR-T 细胞疗法是肝细胞癌（HCC）的一种有前景的治疗方式，但患者间差异显著和抗原异质性限制了其广泛应用。本研究建立了一个基于患者来源类器官（PDO）的平台，用于功能性评估自体 GPC3 靶向 CAR-T 细胞在 HCC 中的活性。HCC PDO 保留了原始肿瘤的关键组织学特征和 GPC3 异质性表达模式。共培养实验中，CAR-T 细胞毒性与 GPC3 表达水平相关，并伴随 IFN-γ 和 IL-2 释放，支持利用 PDO 功能评估 CAR-T 敏感性的可行性。研究还发现，基质条件显著影响类器官结构、病毒转导、CAR-T 细胞浸润和杀伤效率；较低浓度的 Matrigel 更有利于功能评估。值得注意的是，在 GPC3 低表达 PDO 中，以 DNA 甲基转移酶抑制剂 5-氮杂胞苷（5-AZA）预处理可降低 DNA 甲基转移酶 3α（DNMT3A）表达、提高细胞表面 GPC3 表达，并显著增强 CAR-T 介导的细胞毒性。综上，这些发现提供了概念验证证据，支持使用 HCC PDO 作为患者来源平台，模拟部分影响 GPC3 靶向 CAR-T 活性的因素，并探索提高疗效的联合策略。
Glypican-3 (GPC3)-targeted chimeric antigen receptor T (CAR-T) cell therapy is a promising approach for hepatocellular carcinoma (HCC), but marked interpatient variability and antigen heterogeneity limit its broader application. Here, we established a patient-derived organoid (PDO)-based platform to functionally evaluate autologous GPC3-targeted CAR-T cell activity in HCC. HCC PDOs preserved key histologic features and heterogeneous GPC3 expression patterns of the original tumors. In co-culture assays, CAR-T cell cytotoxicity was associated with GPC3 expression levels and was accompanied by IFN- and IL-2 release, supporting the feasibility of using PDOs for functional assessment of CAR-T cell sensitivity. We further found that matrix conditions strongly influenced organoid architecture, viral transduction, CAR-T cell infiltration, and killing efficiency, with lower Matrigel concentrations providing a more permissive setting for functional assessment. Importantly, in GPC3-low PDOs, pretreatment with the DNA methyltransferase inhibitor 5-azacytidine (5-AZA) reduced DNA methyltransferase 3 alpha (DNMT3A) expression, increased surface GPC3 expression, and significantly enhanced CAR-T-mediated cytotoxicity. Together, these findings provide proof-of-concept evidence supporting the use of HCC PDOs as a patient-derived platform for modeling selected determinants of GPC3-targeted CAR-T cell activity and for exploring combination strategies to improve therapeutic efficacy.
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