Fig. 3.

Assessment of allelic status of TP53 lesions. A possibilities of different TP53 configurations and del(17p) presence resulting in monoallelic vs. biallelic lesions. For illustration purpose, four cells scheme is presented. For biallelic mutations, the sum VAF% is more than 50% in hemizygous, homozygous uniparental disomy (UPD) or compound heterozygous configurations. For monoallelic mutations, the sum VAF% is less than 50% in different configurations including heterozygous, hemizygous, or subclonal mosaicism. B The variant allele frequency (VAF) of two TP53 mutations of patients in our cohort was plotted in two colors (red area, obligatory biallelic) with the sum of VAF1 and VAF2 exceeding 50% and (blue area, non-obligatory biallelic) with sum of VAF1 and VAF2 less than 50. C Kaplan–Meier survival estimates of obligatory biallelic, probable biallelic, probable monoallelic, and wild-type TP53 patients after applying different VAF cutoffs based on random forest analysis separating probable monoallelic from probable biallelic mutations. D VAF cutoffs cross-validation by randomly splitting the data into test/train sets with %20/%80 ratios and calculating the Harrell’s C-index (Concordance-index) in the test set over 30 runs. A VAF of 23% resulted optimal for separating the monoallelic and biallelic TP53 mutations. E A novel algorithm for the precise classification of TP53^MT into obligatory biallelic, probable biallelic, or probable monoallelic groups