Journal: Nature Communications

Article Title: A metabolic crosstalk between liposarcoma and muscle sustains tumor growth

doi: 10.1038/s41467-024-51827-3

Figure Lengend Snippet: Real-time qPCR analysis performed on C2C12 cells alone or cocultured with IB115 cells supplemented with vehicle, anti IL-6 (BE8) A , or Siltuximab B , evaluating expression of serine synthesis pathway genes: Phgdh, Psat1 and Psph . (A n = 15, B n = 6, experimental replicates). C , Proliferation assay performed on IB115 cells alone or cocultured with C2C12 cells grown in media without Serine and Glycine and supplemented with vehicle or anti IL-6 (BE8). ( n = 3 experimental replicates). D , Real-time qPCR analysis performed on C2C12 cells alone or cocultured with IB115 cells supplemented with vehicle or GP130 inhibitor (BZA), evaluating expression of serine synthesis pathway genes: Phgdh, Psat1 and Psph . ( n = 15 experimental replicates). E qChIP experiments showing the relative amounts of mono-methylation of lysine 9 of histone H3 (H3K9Me1) on the Psat1 promoter in C2C12 treated or not with recombinant IL-6. Results were represented as the relative ratio between the mean value of immunoprecipitated chromatin (calculated as a percentage of the input) with the indicated antibodies and a control irrelevant antibody. ( n = 3 experimental replicates). F NAD+/NADH ratio in C2C12 treated or not with recombinant IL-6. ( n = 3 experimental replicates). G STAT3, and P-STAT3 protein expression assessed by immunoblot in C2C12 cells incubated 16 h with recombinant IL-6, IB115, or HPAC conditioned media. Tubulin was used as the loading control. H Real-time qPCR analysis performed on C2C12 cells alone or cocultured with IB115 cells (ratio 1:2) supplemented with vehicle or STAT3 inhibitor, evaluating expression of serine synthesis pathway genes: Phgdh, Psat1 , and Psph . ( n = 15 experimental replicates). I Proliferation assay performed on IB115 cells grown in media without Serine and Glycine and cocultured with C2C12 cells treated or not with STAT3 inhibitor. ( n = 3 experimental replicates). Data are shown as means ± SEM and three or more independent experiments were performed. Statistical tests were all adjusted for multiple comparisons and included two-tailed unpaired t-test for (e, f), one-way (a, b, d, h) and two-way analysis of variance (ANOVA) for c and i. * P < 0.05, ** P < 0.01, ***<0.001, n.s. non-significant. Source data are provided as a Source Data file.

Article Snippet: Elsilimomab (BE8) et mAB 1339 (humanized αIl-6) were purchased from Evitria.

Techniques: Expressing, Proliferation Assay, Methylation, Recombinant, Immunoprecipitation, Control, Western Blot, Incubation, Two Tailed Test

Journal: Nature Communications

Article Title: A metabolic crosstalk between liposarcoma and muscle sustains tumor growth

doi: 10.1038/s41467-024-51827-3

Figure Lengend Snippet: A IL6 concentration (ng/ml), measured by ELISA. Control mice were compared to Liposarcoma, Leiomyosarcoma and Breast cancer PDX Mice. ( n = 3 independent experiments). B Real-time qPCR analysis of IL-6 mRNA level in frozen patient samples from different sarcoma subtypes (Adenocarcinoma n = 2, angiosarcoma n = 1, carcinosarcoma n = 3, leiomyosarcoma n = 10, myxoid-liposarcoma n = 7, myxofibrosarcoma n = 7, WDLPS n = 9, DD-LPS n = 3). C Tumor growth curves from patient liposarcoma subcutaneously implanted in nude mice treated or not with Siltuximab (10 mg/kg) or doxorubicine (2 mg/kg) by IP twice weekly. Tumor volume was assessed at the indicated timepoints using caliper measurements. ( n = 6 animals/group). D Tumor growth curves from IB115 LPS cell lines subcutaneously implanted in nude mice treated or not with Siltuximab (10 mg/kg) by IV daily. Tumor volume was assessed at the indicated timepoints using caliper measurements ( n = 3 animals/group). E , IL-6 expression assessed by immunofluorescence performed from IB115 LPS tumors treated or not with Siltuximab (10 mg/kg) by IP twice weekly. ( n = 6 samples/group). F , Tumor growth curves from patient liposarcoma tumor subcutaneously implanted in nude mice, fed a normal or a no Serine/Glycine diet and treated or not with anti-IL-6, BE8 (10 mg/kg) by IP twice weekly after tumor volume reached approximately 150mm 3 . Tumor volume was assessed at the indicated timepoints using caliper measurements ( n = 3 experimental replicates with 6 animals/group). Quantification of serum Serine levels (mM) from Liposarcoma patients ( n = 21 samples/group) G , from Liposarcoma patients (relapsed patients excluded); ( n = 15 samples/group) H before and 30 days after surgery, using liquid chromatography-high resolution mass spectrometry (LC/HRMS). Relapsed patients were highlighted in red. I Quantification of serum Serine levels (mM) from Sarcoma patients before and 30 days after surgery, using liquid chromatography-high resolution mass spectrometry (LC/HRMS). ( n = 38 samples/group). J Schematic representing the crosstalk between liposarcoma and muscle initiated through IL-6/STAT3 pathway activation. Data are shown as means ± SEM and three or more independent experiments were performed. Statistical tests were all adjusted for multiple comparisons and included two-tailed unpaired t-test for (e), one-way (a, b), two-way analysis of variance (ANOVA) (c, d, f) and two-tailed paired and non-parametric Wilcoxon test for g, h and i. * P < 0.05, ** P < 0.01, ***<0.001, n.s non-significant. Source data are provided as a Source Data file.

Article Snippet: Elsilimomab (BE8) et mAB 1339 (humanized αIl-6) were purchased from Evitria.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Control, Expressing, Immunofluorescence, Liquid Chromatography, Mass Spectrometry, Activation Assay, Two Tailed Test