Next-generation RNA sequencing of FFPE subsections reveals highly conserved stromal reprogramming between canine and human mammary carcinoma
Next-generation RNA sequencing of FFPE subsections reveals highly conserved stromal reprogramming between canine and human mammary carcinoma
复制标题
DOI：
10.1242/dmm.040444
复制
发表时间：
2019-08-01
影响因子：
4.3
通讯作者：
Markkanen, Enni
中科院分区：
文献类型：
--
作者：
Amini, Parisa;Nassiri, Sina;Markkanen, Enni
自发性犬单纯性乳腺癌（mCA）通常被视为人类mCA的模型。癌症相关基质（CAS）是人类癌症发生和发展的中心，也可能在犬肿瘤中发挥关键作用。然而，犬CAS缺乏表征，并且尚不清楚犬和人CAS的比较情况。福尔马林固定石蜡包埋（FFPE）组织构成了患者材料的宝贵资源，但化学交联在很大程度上阻碍了下一代RNA测序（RNAseq）对其进行分析。我们最近建立了一个方案，使用激光捕获显微切割，然后使用RNAseq从档案FFPE肿瘤中分离CAS和正常基质。使用这种方法，我们分析了15犬mCA的基质。我们的数据揭示了犬CAS的强重编程。我们证明了犬和人CAS之间的高度分子同源性，并表明上调的犬CAS基因的富集与TCGA乳腺肿瘤数据集中独立衍生的人基质签名的富集强烈相关。在人类乳腺癌中观察到的不同基因特征之间的关系在犬模型中基本保持不变，表明癌症信号通路网络中存在密切的种间相似性。最后，我们建立了犬CAS签名在人类样本中的预后潜力，强调研究犬CAS作为人类疾病模型的相关性。总之，我们提供了通过RNAseq分析FFPE组织的特定亚部分的原理证明，并比较人和犬mCA之间的基质基因表达，以揭示CAS中支持肿瘤生长和恶性的分子驱动因素。
Spontaneous canine simple mammary carcinomas (mCA) are often viewed as models of human mCA. Cancer-associated stroma (CAS) is central for initiation and progression of human cancer, and is likely to play a key role in canine tumours as well. However, canine CAS lacks characterisation and it remains unclear how canine and human CAS compare. Formalin-fixed paraffin embedded (FFPE) tissue constitutes a valuable resource of patient material, but chemical cross linking has largely precluded its analysis by next-generation RNA sequencing (RNAseq). We have recently established a protocol to isolate CAS and normal stroma from archival FFPE tumours using laser-capture microdissection followed by RNAseq. Using this approach, we have analysed stroma from 15 canine mCA. Our data reveal strong reprogramming of canine CAS. We demonstrate a high grade molecular homology between canine and human CAS, and show that enrichment of upregulated canine CAS genes strongly correlates with the enrichment of an independently derived human stromal signature in the TCGA breast tumour dataset. Relationships between different gene signatures observed in human breast cancer are largely maintained in the canine model, suggesting a close interspecies similarity in the network of cancer signalling circuitries. Finally, we establish the prognostic potential of the canine CAS signature in human samples, emphasising the relevance of studying canine CAS as a model of the human disease. In conclusion, we provide a proof-of-principle to analyse specific subsections of FFPE tissue by RNAseq, and compare stromal gene expression between human and canine mCA to reveal molecular drivers in CAS supporting tumour growth and malignancy.