Fig. 6: SHIN1 selectively targets LSCs in combination with standard CML therapy.

a CellTrace Violet (CTV) tracked proliferation of CML CD34⁺ cells following 72 h of treatment with 2 μM IM, 2.5 μM SHIN1 with or without the addition of 1 mM F, or a combination of imatinib and SHIN1, reported as recovery of input (%), where input represents number of input cells (n = 4 patient samples). Relative colony numbers of CML CD34⁺ (b) (n = 7 patient samples) or normal CD34⁺ (c) (n = 3 patient samples) cells following treatment as in (a). Schematics for CML and normal patient samples were created with Biorender.com (Agreement numbers: SH26CFO8OE and IK26CFP2TJ, respectively) d Schematic overview of PDX experimental design: CML CD34⁺ cells were treated with 2 μM imatinib, 2.5 μM SHIN1, or a combination of both for 48 h, and transplanted into sub-lethally irradiated female NRGW⁴¹ mice. After 12 weeks, bone marrow was collected, and engraftment determined by flow cytometry. Created with Biorender.com (Agreement number: ZP26CG12RD). Number of human CD45⁺ (e), CD34⁺ (f), and CD34⁺CD38⁻ (g) CML cells in bone marrow at experimental endpoint (n = 4 mice for untreated group; n = 6 mice for imatinib-treated group; n = 7 mice for SHIN1-treated group, n = 4 mice for combination group). Representative images from treated mice with vehicle, imatinib (50 mg/kg, BID), SHIN1 (100 mg/kg) or a combination of both (h) and quantification (i) of changes in tumour burden as assessed by bioimaging in NRGW⁴¹ mice transplanted with luciferase expressing KCL22 cells following 14 days of treatment (n = 6 mice per group). j Tumour-free survival of mice subjected to treatment is depicted in (h) (n = 6 mice per group). Data are shown as the mean ± s.e.m. P-values are derived from two-way ANOVA with Tukey’s multiple comparisons test (a) relative to untreated CML CD34⁺ cells, ordinary one-way ANOVA with Dunnett’s multiple comparisons test (b, g, i) and log-rank (Mantel–Cox) test for survival analysis (j). Exact p-values in (a) comparing division 3 between UT and SHIN1 treated cells is 0.000052, division 3 between UT and SHIN1 + IM is 0.000001, division 0 between UT and SHIN1 + IM is 0.000007. Source data are provided as a Source Data file.