Fig. 4. Clonal evolution of a multifocal glioblastoma with the TERTp duplication.

a Clinical timeline. Multiple tumor tissue samples were obtained from the IDH-wildtype and TERTp duplicated glioblastoma at diagnosis (SF12747) and one sample was obtained at recurrence (SF13241). b, c MRI of enhancing masses in the left parieto-occipital lobes with surrounding edema and mass effect. d, e H&E staining of an FFPE section from the (d) occipital lobe tumor focus and the (e) parietal lobe tumor focus. Scale bar denotes 50 µm. f TERTp PCR spanning the native ETS motif and duplication for the DNA isolated from FFPE punches from multiple regions of the newly diagnosed tumor, and a no-template control (NTC). Lane 1 includes the DNA ladder for reference of base pair (bp) size. C# and D# indicate the block, # indicate the tumor and N the tumor-adjacent normal brain punch, respectively. The experiment was performed once because of the limitation of patient material. g SNV-based phylogenetic tree from the exome sequencing of DNA from five regions of the tumor and patient-matched normal peripheral blood. Terminal nodes (bold text) represent samples, where N indicates the Normal germline (peripheral blood); all other terminal nodes are tumor samples. Edges (lines) are proportional to the genetic distance between samples within each patient, with the number of mutations corresponding to each major edge indicated. h PyClone analysis of the mutated gene clusters from the exome sequencing of DNA from five regions of the tumor relative to patient-matched normal peripheral blood. N = 1 biologically independent samples for each region. Error bars indicate the mean s.d. (using 10,000 post-‘burn-in’ samples) of Markov chain Monte Carlo–derived cellular prevalence estimates over all mutations in a cluster as defined by PyClone algorithm. TERTp TERT promoter, HD Homozygous deletion, amp Amplification, Dup Duplication. Source data are provided as a Source Data file.