Molecular Pathology Unit, Massachusetts General Hospital Research Institute, Charlestown, MA 02129, USA; Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02139, USA.
Molecular Pathology Unit, Massachusetts General Hospital Research Institute, Charlestown, MA 02129, USA; Massachusetts General Hospital Cancer Center, Harvard Medical School, Charlestown, MA 02129, USA; Center for Regenerative Medicine, Massachusetts General Hospital, Boston, MA 02114, USA; Harvard Stem Cell Institute, Cambridge, MA 02139, USA; Howard Hughes Medical Institute, Bethesda, MD 20815, USA.
Cell. 2019 Jun 13;177(7):1903-1914.e14. doi: 10.1016/j.cell.2019.04.004. Epub 2019 Apr 25.
Xenograft cell transplantation into immunodeficient mice has become the gold standard for assessing pre-clinical efficacy of cancer drugs, yet direct visualization of single-cell phenotypes is difficult. Here, we report an optically-clear prkdc, il2rga zebrafish that lacks adaptive and natural killer immune cells, can engraft a wide array of human cancers at 37°C, and permits the dynamic visualization of single engrafted cells. For example, photoconversion cell-lineage tracing identified migratory and proliferative cell states in human rhabdomyosarcoma, a pediatric cancer of muscle. Additional experiments identified the preclinical efficacy of combination olaparib PARP inhibitor and temozolomide DNA-damaging agent as an effective therapy for rhabdomyosarcoma and visualized therapeutic responses using a four-color FUCCI cell-cycle fluorescent reporter. These experiments identified that combination treatment arrested rhabdomyosarcoma cells in the G2 cell cycle prior to induction of apoptosis. Finally, patient-derived xenografts could be engrafted into our model, opening new avenues for developing personalized therapeutic approaches in the future.
异种细胞移植到免疫缺陷小鼠中已成为评估癌症药物临床前疗效的金标准，但直接观察单细胞表型较为困难。在这里，我们报告了一种光学透明的 prkdc，il2rga 斑马鱼，该斑马鱼缺乏适应性和自然杀伤免疫细胞，可在 37°C 下植入多种人类癌症，并可动态观察单个植入细胞。例如，光转化细胞谱系示踪鉴定了人横纹肌肉瘤（一种儿童肌肉癌）中的迁移和增殖细胞状态。其他实验确定了奥拉帕利（PARP 抑制剂）和替莫唑胺（DNA 损伤剂）联合用药作为横纹肌肉瘤有效治疗方法的临床前疗效，并使用四色 FUCCI 细胞周期荧光报告器观察治疗反应。这些实验表明，联合治疗在诱导细胞凋亡前使横纹肌肉瘤细胞停滞在 G2 细胞周期中。最后，可以将患者来源的异种移植物植入我们的模型中，为未来开发个性化治疗方法开辟了新途径。
