Methodological overview and tomo-seq highlights spatial heterogeneity in 2 whole-tumor lesions. (A) Graphical representation of the patients’ clinical information. Each horizontal line represents a patient. The timeline is segmented and colored according to the duration and treatment agent. The red dot with a cross represents the time at which the biopsies were collected. (B) A scheme illustrating the methods used. (C-D) Longitudinal H&E images of the whole lesion of PT01A (C) and PT10A (D). The horizontal lines represent the approximate locations of the sections used for the tomo-seq. (E-F) Pearson correlation coefficients between every pair of sections calculated using the whole transcriptome profiles for PT01A (E) and PT10A (F). (G-H) Inferred cell type fractions for all sections for PT01A (G) and PT10A (H) generated using CIBERSORT and a reference of RNA sequencing data of pure stroma and immune cell types retrieved from Blueprint and ENCODE. (Gi,Hi) The panel represents all detected cell types, including PCs. (Gii,Hii) Representative fractions of cell types, excluding PCs. (I-J) Normalized counts of GPRC5D and TNFRSF17 in PT01A (I) and PT10A (J). (K) Circos plot of WGS of PT10. A, AraC; Allo SCT, allogeneic stem cell transplantation; B, BCNU; Benda, bendamustine; C, cyclophosphamide; C in PACE, cisplatine; D, doxorubicine; Dara, daratumumab; d/Dex, dexamethasone; E, etoposide; Elo, elotuzumab; F, female; HD, high dose; ide-cel, idecaptagene vicleucel; Ixa, ixazomib; K, carfilzomib; M, male; M/Mel, melphalan; P, prednisone; Pom, pomladomide; R, lenalidomide; T, thalidomide; V, bortezomib.