IntroductionDuring their development, tumour cells accumulate somatic mutations, structural variants and copy number alterations (CNAs). Driver events facilitate clonal expansions and lead to intra-tumour heterogeneity (ITH). While ITH is an important therapeutic challenge, its degree among different cancer types is largely unknown.Material and methodsThe pan-cancer analysis of whole genomes (PCAWG) enabled us to characterise ITH in an unprecedented set of 2778 tumour samples representing 36 histologically distinct cancer types. We applied six CNA callers and eleven subclonal reconstruction algorithms to integrate their solutions into robust consensus copy number profiles and subclonal reconstructions.Results and discussionsOur analysis revealed pervasive ITH in all examined cancer types. We found at least one subclone in 96.7% of the 1801 samples for which we had statistical power to detect subclones. In addition, we find that the average proportions of subclonal point mutations, indels, SVs and CNAs are highly variable across cancer types. These observations suggest distinct evolutionary narratives of each histological cancer type.Analysis of dN/dS ratios shows clear signs of positive selection within both clonal and subclonal mutations. We also identified subclonal mutations in driver genes that are recurrently hit and we found a significant enrichment of subclonal mutations in genes responsible for chromatin regulation. More than 5% of tumours contain driver mutations in genes for which specific treatment is available only in subclones, indicating the importance of assessing the clonality of targeted mutations for clinical decisions.Mutational signatures in the analysed samples show changes in activity over the course of tumour development. Characteristic carcinogen signatures, e.g. UV light exposure in melanomas, make less contributions to subclonal than clonal mutations, while APOBEC-induced mutagenesis has increased activity during the subclonal phase.ConclusionThe absence of a detectable driver mutation in a majority of subclones suggests that late tumour development is frequently driven by CNAs or genomic rearrangements, or that a significant number of late drivers have yet to be identified. We found that selection is widespread and likely the rule rather than the exception and we identified differential activity of mutational signatures, reflecting successive waves of subclonal expansion.