Extended Data Fig. 6: Mutation selection in metastases.

a. Comparison of maximum cancer cell fraction (CCF) in subclonal primary-unique and seeding clusters (Wilcoxon rank-sum test, p = 6.4e-5) and clonal dispersion of primary-unique and seeding clusters (Wilcoxon rank-sum test, p = 1.6e-8). b. Higher dispersion and CCF is seen in the seeding clusters of both primary LN/satellite lesions and recurrence/progression samples compared to non-seeding clusters. Clusters that are found in both primary LN/satellite lesions and recurrence/progression samples were excluded from this analysis. c. Cohort level selection (n genes = 111) of only subclonal mutations in seeding vs. primary-unique mutations vs. mutations in non-metastasizing primary tumours. d. Cohort level selection (n genes = 111) of primary LN/satellite lesions vs. recurrence/progression seeding mutations vs. primary-unique mutations vs. mutations in non-metastasizing primary tumours. Dots represent dN/dS estimates; the asterisks indicate values that are significantly different from 1. e. Gene-level dN/dS values of seeding mutations vs primary-unique and non-metastasizing primary tumour mutations split by lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). Genes are classified as seeding favoured if the odds ratio (OR) of dN/dS of seeding vs. primary-unique mutations >2, primary favoured if OR <0.5, and otherwise classified as both primary and seeding favoured. Genes highlighted in purple and green are significantly enriched in seeding and non-seeding mutations respectively. f. Phylogenetic tree of CRUK0587. Clusters annotated in green are primary-unique, clusters in light purple are shared, while clusters in dark purple are metastasis-unique. There is a metastasis-unique TP53 splice site mutation which occurred independently of a primary-unique S34X TP53 mutation. Lines indicate the 95% confidence intervals for c,d and e. The box plots represent the upper and lower quartiles (box limits), the median (centre line) and the vertical bars span the 5th to 95th percentiles. All tests are two-sided unless otherwise specified.