Extended Data Fig. 1: Loss of ATRX expression is associated with metastasis.
From: Loss of colonic fidelity enables multilineage plasticity and metastasis
(a) International Cancer Genome Consortium data of top 20 mutated cancer genes with high functional impact mutations in colorectal cancer (CRC). (b) Representative ATRX staining of human normal and CRC tissue microarray. Examples of positive and negative staining are shown. Scale bars, 500 µm. (c) Quantification of ATRX expression using immunohistochemistry H-score method analysed using QuPath. Samples are separated into normal, non-metastatic (stage I and II) and metastatic (stage III and IV) groups, n = 8 vs 47 vs 25 tumours. (d) Summary data indicating presence (H-score > 10) or absence (H-score <10) of ATRX staining in non-metastatic and metastatic samples. Number of tumours in each group indicated on graph, n = 47 vs 25 tumours. (e) Summary data indicating presence or absence of ATRX mutation in CRIS-B vs all other CRIS transcriptional subtypes. Data extracted from TCGA dataset where CRIS tumour annotation is known. Number of tumours in each group indicated on graph, n = 43 vs 278 tumours. (f) Overall survival data of patients with CRIS-B tumours separated on presence or absence of ATRX mutation. Data extracted from TCGA dataset, n = 37 vs 6 patients. For (c) data are mean ± SD. P values were calculated using ordinary one-way ANOVA with multiple comparisons. For (d) and (e) p values were calculated using two-sided Fisher’s exact test. For (f) P value was calculated using Log-rank (Mantel-Cox) test. (g) Lollipop plot of TCGA PanCancer mutational data for ATRX. ATRX mutations were analysed using cBioPortal (07/12/23) with TCGA PanCancer Atlas Studies selected. (h) Western-blot analysis of AKP ATRXKO organoids for ATRX and β-actin. n = 2 technical replicates. (i) Representative images of haematoxylin and eosin (H&E) stained lung metastases in mice injected with AKP Control or AKP AtrxKO2 organoid cells via tail vein. Scale bars, 500 µm. (j) Quantification of number of lung metastases per mouse, n = 7 vs 8 mice. (k) Quantification of total lung tumour burden per mouse, n = 7 vs 8 mice. (l) Summary data indicating presence or absence of lung metastases. Number of mice with lung metastases or no metastases indicated on graph, n = 7 vs 8 mice. For (j) and (k) data are mean ± SD. P values were calculated using two-tailed Mann-Whitney test. For (l) p value was calculated using two-sided Fisher’s exact test.
