Journal: Nature Communications

Article Title: PPARα-mediated lipid metabolism reprogramming supports anti-EGFR therapy resistance in head and neck squamous cell carcinoma

doi: 10.1038/s41467-025-56675-3

Figure Lengend Snippet: a Schematic representation for the establishment of cetuximab-sensitive and -resistant patient-derived xenograft (PDX) models from treatment-naive HNSCC clinical specimens. Created in BioRender. https://BioRender.com/b56r875 . Tumor growth of cetuximab-sensitive ( b ) and -resistant ( c ) UCLHN4 PDX in nude mice weekly treated with 30 mg/kg cetuximab or vehicle. Xenografts were considered resistant when tumors reached 200% ±10% of their initial size. d , e Individual GSEA plots of FA metabolism-related pathways in RNA-seq data from cetuximab-sensitive and -resistant UCLHN4 PDX ( N = 3 for each condition). f mRNA expression for CD36 in cetuximab-sensitive and -resistant UCLHN4 PDX ( N = 3). Representative tumor slides ( g ) and quantification ( h ) for anti-CD36 immunohistochemistry on tissue sections from cetuximab-sensitive and -resistant UCLHN4 PDX. Scale bar: 100 µm ( N = 4). Follow-up of tumor growth ( i ) and relative tumor size at day 15 ( j ) for cetuximab-resistant UCLHN4 PDX in nude mice treated with 30 mg/kg cetuximab in combination with either 40 mg/kg etomoxir or 20 mg/kg GW6471. k Schematic representation for the clinical workflow of the window-of-opportunity study with HNSCC patients treated for 14 days with cetuximab. 18 FDG-PET scan was carried out before and after treatment and enabled to discriminate between partial metabolic response (PMR) and stable metabolic disease (SMD). Created in BioRender. https://BioRender.com/o74h220 . l GSEA plot of the PPARα-related gene signature in RNA-seq data from cetuximab-treated HNSCC patients (SMD vs PMR) ( N = 15). Data are plotted as the means ± SEM. N indicates the number of independent biological experiments and n indicates the number of technical replicates (when >1). Significance was determined by two-tailed unpaired Student’s t-test ( h ), one-way ANOVA with Dunnett’s multiple comparison test ( j ) or two-way ANOVA with Sidák’s multiple comparison test ( f ). P -values as indicated or *** P < 0.001. Source data are provided as a Source Data file.

Article Snippet: Briefly, polyA-containing mRNA molecules were purified using poly-T oligo attached magnetic beads from 1 μg total RNA (with the Magnetic mRNA Isolation Kit; New England Biolabs), before fragmentation using divalent cations (under elevated temperature to obtain approximately 300 bp pieces), double strand cDNA synthesis, and finally Illumina adapter ligation and cDNA library amplification by PCR for sequencing.

Techniques: Derivative Assay, RNA Sequencing Assay, Expressing, Immunohistochemistry, Two Tailed Test, Comparison