Application of high-throughput single-nucleus DNA sequencing in pancreatic cancer

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Abstract
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.
Funding Information
  • Mochida Memorial Foundation for Medical and Pharmaceutical Research
  • Daiichi Sankyo Foundation of Life Science
  • U.S. Department of Health & Human Services | NIH | National Cancer Institute (F31 CA260796-01, K99/R00 CA229979-01A1)
  • U.S. Department of Health & Human Services | NIH | National Cancer Institute
  • U.S. Department of Health & Human Services | National Institutes of Health (CA220508)

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