Post-mortem Plasma Cell-Free DNA Sequencing: Proof-of-Concept Study for the "Liquid Autopsy".
Post-mortem Plasma Cell-Free DNA Sequencing: Proof-of-Concept Study for the "Liquid Autopsy".
复制标题
死后血浆无细胞 DNA 测序：“液体尸检”的概念验证研究。
DOI：
10.1038/s41598-020-59193-y
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Goto A.
中科院分区：
文献类型：
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作者：
Ito Y;Yoshida M;Inoue T;Habuchi T;Ikoma S;Katoh H;Kato M;Shibata T;Ishikawa S;Yachida S;Goto A.
最近的基因组研究在快速尸检过程中获得的癌症组织提供了深入了解克隆进化和异质性的癌症。然而，死后血液尚未进行与癌症有关的基因分析。我们首先证实，大量的细胞游离DNA存在于12例尸检的死后血浆。然后，我们集中在一个试点的情况下，前列腺癌与多个转移的基因分析。对快速尸检过程中收集的死后血浆来源的细胞游离DNA和8个冷冻转移癌组织进行全外显子组测序，并比较它们的突变状态。对死后血浆游离DNA成功测序，鉴定出344个突变。其中，160个在至少一个转移灶中检测到。此外，所有转移瘤共有的99%的突变存在于血浆中。桑格测序的30个额外的福尔马林固定转移使我们能够映射的克隆携带突变最初只在血浆中检测到。总之，死后的血液，这通常是在传统的尸检处理，可以提供有价值的数据，如果详细测序，特别是关于癌症的异质性。此外，死后血浆无细胞DNA测序（液体尸检）可以成为癌症研究的新平台和基因组病理学的工具。
Recent genomic studies on cancer tissues obtained during rapid autopsy have provided insights into the clonal evolution and heterogeneity of cancer. However, post-mortem blood has not been subjected to genetic analyses in relation to cancer. We first confirmed that substantial quantities of cell-free DNA were present in the post-mortem plasma of 12 autopsy cases. Then, we focused on a pilot case of prostate cancer with multiple metastases for genetic analyses. Whole-exome sequencing of post-mortem plasma-derived cell-free DNA and eight frozen metastatic cancer tissues collected during rapid autopsy was performed, and compared their mutational statuses. The post-mortem plasma cell-free DNA was successfully sequenced and 344 mutations were identified. Of these, 160 were detected in at least one of the metastases. Further, 99% of the mutations shared by all metastases were present in the plasma. Sanger sequencing of 30 additional formalin-fixed metastases enabled us to map the clones harboring mutations initially detected only in the plasma. In conclusion, post-mortem blood, which is usually disposed of during conventional autopsies, can provide valuable data if sequenced in detail, especially regarding cancer heterogeneity. Furthermore, post-mortem plasma cell-free DNA sequencing (liquid autopsy) can be a novel platform for cancer research and a tool for genomic pathology.