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. 2023 Aug 17;72(12):2294–2306. doi: 10.1136/gutjnl-2022-329140

Figure 6.

Figure 6

Small molecule inhibition of NOTUM inhibits tumour growth and metastasis in a mouse model of colorectal cancer. (A) Brightfield image of Apc mutant and APKS mouse tumouroids treated with the small molecule inhibitor of NOTUM, ABC99 or vehicle control (DMSO) (n=3 technical). Scale bar: 100 µm. (B) Single cell clonal seeding efficiency and EdU assay quantifying fraction of cells in S phase from tumouroids in (A) (n=3 technical replicates). (C) Bioluminescent imaging (BLI) of mice after endoscope-guided orthotopic implantation of APKS mouse tumouroids into the colonic mucosa. Mice were treated with ABC99 or vehicle (corn oil) control after 4 weeks of engraftment and tumour growth. Mice are treated for indicated number of weeks (week 0=4 weeks post-implantation) (n=7 mice). (D) Quantification of luminescence radiance from (C) at indicated time points (n=7 mice). (E–G) Quantification of primary tumour weight (E), number of macrometastases in the liver (F), and number of lymph node metastases (G) in mice from (C) at the termination of the experiment (8 weeks total, 4 weeks post-treatment). (H) Immunofluorescence staining for KI67 and E-CADHERIN in primary tumour and liver metastases from tumours in (C) (n=7 mice). Nuclei are visualised by DAPI staining. Scale bar of top panels: 200 µm. Lower panels: 50 µm. (I) Kaplan-Meier survival analysis for mice harbouring orthotopic APKS tumours as in (C), treated with ABC99 or vehicle (corn oil) control for 16 weeks following an initial 4-week period of engraftment and tumour growth (total experimental time course of 5 months) (n=10 mice). For all panels, **p<0.01, ***p<0.001, Student’s t-test.