Figure 6: Clonal evolution from MDS to AML in RCMD-RS patient with SF3B1 mutation.

(a) FACS profiles of stem cell progenitors in BM MNCs in a serial sample at the MDS stage and AML stage of the disease. (b) Interphase FISH for tracking the del(7q) aberration in cells obtained at the AML stage of the disease. Two chromosome-7 aberrations, del(7q36) and del(7q22-7q36), were detected as two separate clones. Two individual cells are shown in the figure with one harbouring del(7q36) represented by a green probe and the other cell has del(7q22-7q36) represented by probes orange/green. (c) SNV profile derived from whole-exome sequencing. The shown profile depicts a heterozygous deletion (horizontal red line) of chromosome 7q in patient CD34⁺ bone marrow cells (AML stage, bottom) and absence of the deletion in paired control CD3⁺ cells (top). (d) Bar chart (FISH analysis) showing percentage of CD34⁺ cells (MDS stage), CD34⁺ cells (AML stage), MPL (AML stage), GMP (AML stage) with del(7q) aberration. (e) Bar chart (FISH analysis) showing the percentage of del(7q) aberration in LTC-derived cells from MDS-stage CD34⁺ cells (time point 1). (f) Mutational status of the gene mutations in sequential bone marrow samples (MDS-stage time-point 1, MDS-stage time-point 2 and AML stage). Coloured circles represent the MABs for each screened gene mutation. Black * represent absence of gene mutations in the AML-stage sample. Red * represent acquisition of additional mutation at the AML stage. (g) Mutational analysis of SF3B1, DNMT3A and SUV420H1 in HSCs, MLPs and GMPs from the AML stage of the disease. Sequencing depth for HSCs was >10,000 reads. (h) Proposed sequential acquisition of genetic lesions based on analysis of sequential samples, LTC-derived cells and single-cell clonogenic assays.