Vick Binje, Rothenberg Maja, Sandhöfer Nadine, Carlet Michela, Finkenzeller Cornelia, Krupka Christina, Grunert Michaela, Trumpp Andreas, Corbacioglu Selim, Ebinger Martin, André Maya C, Hiddemann Wolfgang, Schneider Stephanie, Subklewe Marion, Metzeler Klaus H, Spiekermann Karsten, Jeremias Irmela
Group Apoptosis, Department of Gene Vectors, Helmholtz Zentrum München, German Research Center for Environmental Health, Munich, Germany; German Cancer Consortium (DKTK), Heidelberg, Germany; German Cancer Research Center (DKFZ), Heidelberg, Germany.
Department of Internal Medicine III, University Hospital Grosshadern, Ludwig-Maximilians-Universität (LMU), Munich, Germany.
PLoS One. 2015 Mar 20;10(3):e0120925. doi: 10.1371/journal.pone.0120925. eCollection 2015.
Acute myeloid leukemia (AML) is a clinically and molecularly heterogeneous disease with poor outcome. Adequate model systems are required for preclinical studies to improve understanding of AML biology and to develop novel, rational treatment approaches. Xenografts in immunodeficient mice allow performing functional studies on patient-derived AML cells. We have established an improved model system that integrates serial retransplantation of patient-derived xenograft (PDX) cells in mice, genetic manipulation by lentiviral transduction, and essential quality controls by immunophenotyping and targeted resequencing of driver genes. 17/29 samples showed primary engraftment, 10/17 samples could be retransplanted and some of them allowed virtually indefinite serial transplantation. 5/6 samples were successfully transduced using lentiviruses. Neither serial transplantation nor genetic engineering markedly altered sample characteristics analyzed. Transgene expression was stable in PDX AML cells. Example given, recombinant luciferase enabled bioluminescence in vivo imaging and highly sensitive and reliable disease monitoring; imaging visualized minimal disease at 1 PDX cell in 10000 mouse bone marrow cells and facilitated quantifying leukemia initiating cells. We conclude that serial expansion, genetic engineering and imaging represent valuable tools to improve the individualized xenograft mouse model of AML. Prospectively, these advancements enable repetitive, clinically relevant studies on AML biology and preclinical treatment trials on genetically defined and heterogeneous subgroups.
急性髓系白血病（AML）是一种临床和分子层面均具有异质性且预后较差的疾病。临床前研究需要合适的模型系统，以增进对AML生物学的理解，并开发新的合理治疗方法。免疫缺陷小鼠中的异种移植可对患者来源的AML细胞进行功能研究。我们建立了一种改进的模型系统，该系统整合了小鼠体内患者来源异种移植（PDX）细胞的连续再移植、慢病毒转导的基因操作以及通过免疫表型分析和驱动基因靶向重测序进行的基本质量控制。29个样本中有17个显示出初次植入，17个样本中有10个可以进行再移植，其中一些样本几乎可以无限期连续移植。6个样本中有5个使用慢病毒成功转导。连续移植和基因工程均未明显改变所分析的样本特征。转基因在PDX AML细胞中表达稳定。举例来说，重组荧光素酶可实现体内生物发光成像以及高度灵敏且可靠的疾病监测；成像可在10000个小鼠骨髓细胞中的1个PDX细胞水平上检测到微小疾病，并有助于对白血病起始细胞进行定量分析。我们得出结论，连续扩增、基因工程和成像技术是改进AML个体化异种移植小鼠模型的重要工具。展望未来，这些进展能够使我们对AML生物学进行重复性的、与临床相关的研究，并针对基因定义的异质性亚组开展临床前治疗试验。
