Extended Data Fig. 8: Spatial proteomic profiling of Richter’s Transformation.
a Representative high-resolution images of DLBCL (top) and CLL (bottom) regions showing differences in cell size (n = 1). b Schematic of the DBiTplus workflow using CellScape on an FFPE tissue section. Created with BioRender.com. c Protein marker panel used in CellScape, including immune, stromal, and segmentation markers. d Stacked barplot showing cell type composition in CLL and DLBCL regions. e Boxplot where the central line represents the median, the bottom and upper edges of the box indicate the first and third quartiles, and the whiskers extend to the most extreme data points within 1.5 times the interquartile range (IQR) from the box edge for the quantification of average T cell-tumor cell distances in DLBCL compared to CLL. Computed using a two-tailed Mann-Whitney test (**** represents p < 0.0001, nCLL = 5,525 and nDLBCL = 4,924). Median distance 7.715 µm and 7.292 µm, respectively for DLBCL and CLL. f Violin plots of the distance of T cell subtypes from tumor cells in DLBCL versus CLL regions. Violin plots show the median (center line), the first and third quartiles (box limits) and 1.5x interquartile range (whiskers). g Boxplot, where the central line represents the median, the bottom and upper edges of the box indicate the first and third quartiles, and the whiskers extend to the most extreme data points within 1.5 times the interquartile range (IQR) from the box edge showing size distribution across all major annotated cell types. h Histogram of cell size distributions comparing large B cells (DLBCL) and small B cells (CLL). Histogram bin width of 0.25 i Spatial maps showing localization of large B cells (red) and small B cells (blue) within the tissue section (left), and CLL cells only (right). j, 2D density scatter plot showing CD20 and Ki67 expression as a function of cell size in large B cells (left) versus other B cells (right). Small B cells < 300 in size and < 0.30 in normalized Ki67 expression were annotated as CLL-like cells. k, Density distribution plots of marker expression (CD23, CD5, LEF1) in small (top) and large B cells (bottom). l Spatial maps showing localization of CLL-like cells.
