Division of Children's Leukaemia and Cancer Research, Telethon Institute for Child Health Research, Perth, Western Australia.
BMC Genomics. 2010 Apr 21;11:256. doi: 10.1186/1471-2164-11-256.
Pre-clinical models that effectively recapitulate human disease are critical for expanding our knowledge of cancer biology and drug resistance mechanisms. For haematological malignancies, the non-obese diabetic/severe combined immunodeficient (NOD/SCID) mouse is one of the most successful models to study paediatric acute lymphoblastic leukaemia (ALL). However, for this model to be effective for studying engraftment and therapy responses at the whole genome level, careful molecular characterisation is essential.
Here, we sought to validate species-specific gene expression profiling in the high engraftment continuous ALL NOD/SCID xenograft. Using the human Affymetrix whole transcript platform we analysed transcriptional profiles from engrafted tissues without prior cell separation of mouse cells and found it to return highly reproducible profiles in xenografts from individual mice. The model was further tested with experimental mixtures of human and mouse cells, demonstrating that the presence of mouse cells does not significantly skew expression profiles when xenografts contain 90% or more human cells. In addition, we present a novel in silico and experimental masking approach to identify probes and transcript clusters susceptible to cross-species hybridisation.
We demonstrate species-specific transcriptional profiles can be obtained from xenografts when high levels of engraftment are achieved or with the application of transcript cluster masks. Importantly, this masking approach can be applied and adapted to other xenograft models where human tissue infiltration is lower. This model provides a powerful platform for identifying genes and pathways associated with ALL disease progression and response to therapy in vivo.
能够有效重现人类疾病的临床前模型对于扩展我们对癌症生物学和耐药机制的认识至关重要。对于血液系统恶性肿瘤,非肥胖型糖尿病/严重联合免疫缺陷(NOD/SCID)小鼠是研究小儿急性淋巴细胞白血病(ALL)的最成功模型之一。然而,为了使该模型能够有效地在全基因组水平上研究植入和治疗反应,必须进行仔细的分子特征描述。
在这里,我们试图在高植入性连续 ALL NOD/SCID 异种移植中验证种属特异性基因表达谱。我们使用人类 Affymetrix 全转录体平台分析了未经过细胞分离的植入组织的转录谱,发现它可以在单个小鼠的异种移植中返回高度可重复的谱。该模型进一步通过人源和鼠源细胞的实验混合物进行了测试,证明当异种移植中含有 90%或更多的人源细胞时,鼠源细胞的存在不会显著扭曲表达谱。此外,我们提出了一种新的基于计算和实验的掩蔽方法,用于鉴定易受种间杂交影响的探针和转录簇。
我们证明了当达到高植入水平或应用转录簇掩蔽时,可以从异种移植中获得种属特异性的转录谱。重要的是,这种掩蔽方法可以应用于其他人类组织浸润程度较低的异种移植模型中。该模型为鉴定与 ALL 疾病进展和体内治疗反应相关的基因和途径提供了一个强大的平台。