PO.TB04.03 · 肿瘤生物学
利用肿瘤和胃肠道类器官模型对HER2 CAR-T细胞进行临床前评估以界定治疗窗
Preclinical assessment of HER2 CAR-T cells using tumor and GI organoid models to define therapeutic window
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
抗HER2疗法已显著改善了HER2+肿瘤转移性乳腺癌患者的结局。除trastuzumab等基于抗体的方法外,HER2靶向CAR-T细胞正成为实体瘤有前景的选择。然而,癌症疗法的临床转化常受脱靶毒性的阻碍,尤其是在胃肠道(GI)中。CAR-T细胞可引起上皮损伤,导致腹泻和黏膜炎症等不良事件。为解决这一问题,我们采用三维iPSC来源的肠道类器官(HIOs)作为一个具有人体生理相关性的模型来评估GI毒性。通过活力实验、切割型caspase 3和7,我们评估了与PBMCs共培养的HIOs中直接细胞毒性以及免疫介导的上皮损伤。同时,我们在HER2+和HER2−人癌细胞系(SK-OV3、JIMT-1、Hs578T)的二维和三维体外模型中,考察了HER2靶向CAR-T细胞相较于作为阳性对照的trastuzumab的疗效。通过基于荧光的活细胞成像测定CAR-T细胞的肿瘤生长和侵袭。在实验的最后一天,进行了代谢读出(CellTiter-Glo,CTG实验)。CAR-T细胞对HER2+球体(包括trastuzumab抵抗的JIMT-1)表现出强效的、剂量依赖性的细胞毒性,同时不影响HER2− Hs578T细胞。Trastuzumab仅对SK-OV3有效。未转导的T细胞则被证明无效。同时,CAR-T细胞在HIOs中诱导了上皮损伤,提示在后续药物开发阶段可能存在的安全性问题。此外,在该实验中,来自同一供体的未转导T细胞未诱导任何细胞毒性。总之,将基于肠道类器官的毒性筛选与肿瘤疗效实验相整合,能够更精确地界定HER2 CAR-T细胞的治疗窗。这种双平台方法通过同时捕捉疗效和GI安全性,增强了临床前评估,支持开发具有更佳转化潜力的细胞疗法。
查看英文原文 English abstract
Anti-HER2 therapies have significantly improved outcomes for metastatic breast cancer patients with HER2+ tumors. Beyond antibody-based approaches such as trastuzumab, HER2-targeted CAR-T cells are emerging as promising options for solid tumors. However, the clinical translation of cancer therapies is often hindered by off-target toxicities, particularly in the gastrointestinal (GI) tract. CAR-T cells, can cause epithelial damage, leading to adverse events such as diarrhea and mucosal inflammation. To address this, we employed 3D iPSC-derived intestinal organoids (HIOs) as a human physiologically relevant model to assess GI toxicity. Using viability assays, cleaved caspase 3 and 7, we evaluated both direct cytotoxic and immune-mediated epithelial injury in HIOs co-cultured with PBMCs. In parallel, we investigated the efficacy of HER2-targeting CAR-T cells versus trastuzumab as positive control in HER2+ and HER2− human cancer cell lines (SK-OV3, JIMT-1, Hs578T) in 2D and 3D in vitro models. Tumor growth and invasion of the CAR-T cells was measured via fluorescence-based live cell imaging. On the last experiment day, a metabolic read-out (CellTiter-Glo, CTG assay) was performed. CAR-T cells demonstrated potent, dose-dependent cytotoxicity against HER2+ spheroids, including trastuzumab-resistant JIMT-1, while sparing HER2− Hs578T cells. Trastuzumab was effective only in SK-OV3. The un-transduced T cells proved to be ineffective. In parallel, the CAR T cells induced epithelial injury in the HIOs indicating possible safety concerns in later drug development stages. Also, in this assay the untransduced T cells from the same donor did not induce any cytotoxicity. In summary, the integration of intestinal organoid-based toxicity screening with tumor efficacy assays enabled a more precise definition of the therapeutic window for HER2 CAR-T cells. This dual-platform approach enhances preclinical evaluation by simultaneously capturing efficacy and GI safety, supporting the development of cell therapies with improved translational potential.
利益披露 Disclosure
J. B. Schueler, None..
I. Rohleff, None..
T. Martianez Canales, None..
K. Lashuk, None..
Y. von Elsswilk, None..
N. Jayanth, None..
L. Buti, None.