Representative Sequencing: Unbiased Sampling of Solid Tumor Tissue.
Although thousands of solid tumors have been sequenced to date, a fundamental under-sampling bias is inherent in current methodologies. This is caused by a tissue sample input of fixed dimensions (e.g., 6 mm biopsy), which becomes grossly under-powered as tumor volume scales. Here, we demonstrate representative sequencing (Rep-Seq) as a new method to achieve unbiased tumor tissue sampling. Rep-Seq uses fixed residual tumor material, which is homogenized and subjected to next-generation sequencing. Analysis of intratumor tumor mutation burden (TMB) variability shows a high level of misclassification using current single-biopsy methods, with 20% of lung and 52% of bladder tumors having at least one biopsy with high TMB but low clonal TMB overall. Misclassification rates by contrast are reduced to 2% (lung) and 4% (bladder) when a more representative sampling methodology is used. Rep-Seq offers an improved sampling protocol for tumor profiling, with significant potential for improved clinical utility and more accurate deconvolution of clonal structure.
['Litchfield|Kevin|K|', 'Stanislaw|Stacey|S|', 'Spain|Lavinia|L|', 'Gallegos|Lisa L|LL|', 'Rowan|Andrew|A|', 'Schnidrig|Desiree|D|', 'Rosenbaum|Heidi|H|', 'Harle|Alexandre|A|', 'Au|Lewis|L|', 'Hill|Samantha M|SM|', 'Tippu|Zayd|Z|', 'Thomas|Jennifer|J|', 'Thompson|Lisa|L|', 'Xu|Hang|H|', 'Horswell|Stuart|S|', 'Barhoumi|Aoune|A|', 'Jones|Carol|C|', 'Leith|Katherine F|KF|', 'Burgess|Daniel L|DL|', 'Watkins|Thomas B K|TBK|', 'Lim|Emilia|E|', 'Birkbak|Nicolai J|NJ|', 'Lamy|Philippe|P|', 'Nordentoft|Iver|I|', 'Dyrskjøt|Lars|L|', 'Pickering|Lisa|L|', 'Hazell|Stephen|S|', 'Jamal-Hanjani|Mariam|M|', '|||', 'Larkin|James|J|', 'Swanton|Charles|C|', 'Alexander|Nelson R|NR|', 'Turajlic|Samra|S|']
D014408:Biomarkers, Tumor D001706:Biopsy D059014:High-Throughput Nucleotide Sequencing D006801:Humans D008175:Lung Neoplasms D009154:Mutation D047368:Tumor Burden D001749:Urinary Bladder Neoplasms
homogenization tumor mutational burden representative sampling biomarkers molecular profiling tumor hetereogeneity tumor sampling tumor sequencing