Extended Data Figure 5: CD36+ cells are defined by a lipid metabolism and metastatic signature, and require the fatty acid β-oxidation enzyme ACSL1 to promote metastasis.
From: Targeting metastasis-initiating cells through the fatty acid receptor CD36
a, b, Top categories for diseases (a) and biological process (b) upregulated in CD36+ CD44bright cells. c, Gene set enrichment analysis (GSEA) plot of CD36-associated signatures, highlighting strong enrichment for fatty acid metabolism. NES denotes normalized enrichment score. d, Comparative analysis of overlapping genes between CD36+ CD44bright and CD44bright DID+ upregulated signature, highlighting over-represented genes associated with lipid metabolism, cancer invasion and metastasis and transport and metabolism of nucleoside drugs. P = 1.359 × 10−16, hypergeometric test. e, Flow cytometry analysis of in vitro SCC-25 cells co-cultured with adipogenic OP-9 cells, showing the expression of three enzymes of fatty acid β-oxidation (ACADVL, ACADM and HADHA). Histograms show the average normalized number of events as a function of fluorescence intensity for the three enzymes (n = 2 biological replicates). f, BLI monitoring of tumours generated from OSCC cells transduced with either scrambled shRNA (SCR, n = 5 mice) or shRNA ACSL1#936 (n = 5 mice). Graphs show the frequency of developed tumours and the BLI signal quantification (**P = 0.001 and *P = 0.003, two-tailed t-test). g, Haematoxylin and eosin staining of metastatic lymph nodes from animals reported in f, showing the smaller metastases arising from Acsl1 shRNA transplants as compared to the control SCR (n = 5 animals per group). h, BLI monitoring of orthotopic transplants from CD36-overexpressing JHU-029 cells transduced with either control (SCR, n = 10 mice) or shRNA ACSL1#936 (n = 10 mice). Graphs show the BLI signal quantification (metastasis: *P = 0.03 and *P = 0.03, two-tailed t-test) and the frequency of developed tumours (CT vs OE-SCR *P = 0.03 and OE-SCR vs OE-shACSL1 *P = 0.04, Fisher exact test). i, Histogram shows the average normalized number of events as a function of CD36 fluorescence intensity. j, Relative RNA levels of OSCC cells reported in j, by RT–qPCR analysis. Human β-2-microglobulin was used as internal control gene (n = 3 biological replicates, P = 0.03, two-tailed t-test). Data in f, h, j, are given as the mean ± s.e.m. Source data from mouse experiments are in Supplementary Information.
