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Application of high-throughput single-nucleus DNA sequencing in pancreatic cancer
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Application of high-throughput single-nucleus DNA sequencing in pancreatic cancer
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Application of high-throughput single-nucleus DNA sequencing in pancreatic cancer
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Journal Article
Application of high-throughput single-nucleus DNA sequencing in pancreatic cancer
2023
Overview
Despite insights gained by bulk DNA sequencing of cancer it remains challenging to resolve the admixture of normal and tumor cells, and/or of distinct tumor subclones; high-throughput single-cell DNA sequencing circumvents these and brings cancer genomic studies to higher resolution. However, its application has been limited to liquid tumors or a small batch of solid tumors, mainly because of the lack of a scalable workflow to process solid tumor samples. Here we optimize a highly automated nuclei extraction workflow that achieves fast and reliable targeted single-nucleus DNA library preparation of 38 samples from 16 pancreatic ductal adenocarcinoma patients, with an average library yield per sample of 2867 single nuclei. We demonstrate that this workflow not only performs well using low cellularity or low tumor purity samples but reveals genomic evolution patterns of pancreatic ductal adenocarcinoma as well.
Implementing high-throughput single-cell DNA sequencing for the study of solid tumours has been challenging. Here, the authors present an optimised approach for snap-frozen tissue single nuclei extraction and DNA sequencing, which can be applied to study pancreatic ductal adenocarcinoma evolution and heterogeneity.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 45/23
/ 45/62
/ Cancer
/ Carcinoma, Pancreatic Ductal - genetics
/ Carcinoma, Pancreatic Ductal - pathology
/ DNA
/ Genomics
/ High-Throughput Nucleotide Sequencing
/ Humanities and Social Sciences
/ Humans
/ Nuclei
/ Pancreatic Neoplasms - genetics
/ Pancreatic Neoplasms - pathology
/ Science
/ Tumors
/ Workflow
