Genomic evolution and clonal dynamics of pancreatic cancer at single-nucleus resolution

Key Methodology: High-Throughput Targeted snDNA-seq by Tapestri
The Mission Bio Tapestri platform was utilized to resolve single-cell copy number alterations (CNAs) and single-nucleotide variants (SNVs) simultaneously across multi-region and longitudinal tissue samples.

Method: Researchers applied an optimized archival single-nucleus DNA sequencing workflow to 24 patient cases (including surgical resections and rapid autopsies). Single nuclei were isolated, microfluidically encapsulated on the Tapestri platform, and targeted multiplex PCR was performed to generate high-coverage sequencing libraries.

Detection & Benefits: Tapestri allowed the accurate reconstruction of phylogenetic trees by distinguishing co-occurring versus mutually exclusive mutations. It revealed that copy number variations (CNVs) account for the vast majority of spatial tumor heterogeneity and uncovered hidden subclonal resistance mechanisms to KRAS inhibition. Additionally, the single-cell resolution exposed varied biallelic inactivation mechanisms of BRCA2 and the precise evolutionary timing of TGF-β pathway alterations during tissue invasion.

Read more