Single-cell sequencing defines genetic heterogeneity in pancreatic cancer precursor lesions
Single-cell sequencing defines genetic heterogeneity in pancreatic cancer precursor lesions
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DOI：
10.1002/path.5194
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发表时间：
2019-03-01
影响因子：
7.3
通讯作者：
Wood, Laura D.
中科院分区：
文献类型：
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作者：
Kuboki, Yuko;Fischer, Catherine G.;Wood, Laura D.
导管内乳头状黏液性肿瘤(IPMN)是胰腺癌的先兆，然而，对这些病变的遗传异质性知之甚少。这项研究的目的是在单细胞水平上表征IPMN的遗传异质性。我们从10个IPMN的新鲜组织中分离出单个细胞，然后进行全基因组扩增和下一代胰腺驱动基因的定向测序。然后，我们使用一种新的多样本突变调用算法来确定单细胞基因类型。我们的分析表明，同一驱动基因的不同突变经常发生在同一IPMN中。两个IPMN在启动驱动基因KRAS中有多个突变，这些突变发生在独特的肿瘤克隆中，这表明可能是多克隆起源或在这个关键突变之前发生了不明的启动事件。较晚发生的驱动基因的多重突变也很常见，并经常定位于独特的肿瘤克隆，增加了这些基因事件在胰腺肿瘤发生中收敛进化的可能性。IPMN的单细胞测序显示了早期和晚期发生的驱动基因突变的遗传异质性，这表明在这些病变中，肿瘤进化的模式比以前所认识的更复杂。版权所有(C)2018年大不列颠和爱尔兰病理学会。作者：John Wiley&Sons，Ltd.
Intraductal papillary mucinous neoplasms (IPMNs) are precursors to pancreatic cancer; however, little is known about genetic heterogeneity in these lesions. The objective of this study was to characterize genetic heterogeneity in IPMNs at the single-cell level. We isolated single cells from fresh tissue from ten IPMNs, followed by whole genome amplification and targeted next-generation sequencing of pancreatic driver genes. We then determined single-cell genotypes using a novel multi-sample mutation calling algorithm. Our analyses revealed that different mutations in the same driver gene frequently occur in the same IPMN. Two IPMNs had multiple mutations in the initiating driver gene KRAS that occurred in unique tumor clones, suggesting the possibility of polyclonal origin or an unidentified initiating event preceding this critical mutation. Multiple mutations in later-occurring driver genes were also common and were frequently localized to unique tumor clones, raising the possibility of convergent evolution of these genetic events in pancreatic tumorigenesis. Single-cell sequencing of IPMNs demonstrated genetic heterogeneity with respect to early and late occurring driver gene mutations, suggesting a more complex pattern of tumor evolution than previously appreciated in these lesions. Copyright (c) 2018 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.