Journal: Cancer Immunology Research

Article Title: IL1R2 Deficiency Unleashes Neutrophil-Mediated Antitumor Potential in Sarcoma

doi: 10.1158/2326-6066.CIR-25-0651

Figure Lengend Snippet: Cellular mechanisms of resistance to sarcoma growth in IL1R2-deficient mice. A and B, FS6 tumor growth curve (left) or primary tumor volume at the time of sacrifice (right) in Il1r2 +/+ ( n = 24), Il1r2 −/− ( n = 23), Csf3r −/− ( n = 22), and Il1r2 −/− /Csf3r −/− ( n = 19) mice ( A ) and analysis by FACS of neutrophil frequency on CD45 + cells in the spleen and tumor of sarcoma-bearing mice expressing Il1r2 +/+ ( n = 7), Il1r2 −/− ( n = 5), Csf3r −/− ( n = 6), and Il1r2 −/− /Csf3r −/− ( n = 4; B ). C–E, FS6 tumor volume ( C–E ) and growth curve ( E ) in Il1r2 +/+ and Il1r2 −/− mice treated with anti-CD8 and/or anti-NK1.1 ( C, n = 9 Il1r2 +/+ mice + ctrl IgG, n = 9 Il1r2 +/+ mice + anti-CD8, n = 8 Il1r2 −/− mice + ctrl IgG, and n = 6 Il1r2 −/− mice + anti-CD8; D, n = 8 Il1r2 +/+ mice + ctrl IgG, n = 9 Il1r2 +/+ mice + anti-NK1.1, n = 8 Il1r2 −/− mice + ctrl IgG, and n = 7 Il1r2 −/− mice + anti-NK1.1; E, n = 7 Il1r2 +/+ mice + ctrl IgG, n = 8 Il1r2 +/+ mice + anti–CD8–NK1.1, n = 6 Il1r2 −/− mice + ctrl IgG, and n = 6 Il1r2 −/− mice + anti–CD8–NK1.1). F and G, FS6 tumor growth curve in Il1r2 +/+ and Il1r2 −/− mice treated with anti–PD-1 ( F ) or anti-IFNγ ( G ; F, n = 5 Il1r2 +/+ mice + ctrl IgG, n = 6 Il1r2 +/+ mice + anti–PD-1, n = 6 Il1r2 −/− mice + ctrl IgG, and n = 7 Il1r2 −/− mice + anti–PD-1; G, n = 5 Il1r2 +/+ mice + ctrl IgG, n = 8 Il1r2 +/+ mice + anti-IFNγ, n = 7 Il1r2 −/− mice + ctrl IgG, and n = 9 Il1r2 −/− mice + anti-IFNγ). H and I, FS6 primary tumor volume in WT ( n = 16), Il1r2 −/− ( n = 5), and Il1r1 −/− ( n = 7) mice ( H ) and analysis by FACS of neutrophil frequency on CD45 + cells in the blood, spleen, and tumor of WT ( n = 10), Il1r2 −/− ( n = 5), and Il1r1 −/− ( n = 5) sarcoma-bearing mice ( I ). J, FS6 tumor growth curve in Il1r2 −/− mice treated with vehicle ( n = 9) or anakinra ( n = 11). Mean ± SEM. Three pooled experiments ( A ) and one experiment ( B–J ) performed. Experiments of panels H and I were performed using a pool of commercial and littermate WT mice that showed similar tumor growth and neutrophil frequency. P values determined by two-way ANOVA [ A and E (left), F–H , and J ] or two-tailed Mann–Whitney U test [ A and E (right), B–D , and I ]. ns, not statistically significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Il1r1 −/− (RRID: IMSR_JAX:003245) and Csf3r −/− (RRID: IMSR_JAX:017838) mice were purchased from The Jackson Laboratory.

Techniques: Expressing, Two Tailed Test, MANN-WHITNEY

Journal: Cancer Immunology Research

Article Title: IL1R2 Deficiency Unleashes Neutrophil-Mediated Antitumor Potential in Sarcoma

doi: 10.1158/2326-6066.CIR-25-0651

Figure Lengend Snippet: Phenotype of neutrophils from sarcoma-bearing IL1R2-deficient mice. A and B, Analysis by FACS of CD101 + cell frequency on total neutrophils ( A ) or frequency of CD101 + and CD101 − neutrophil subsets on total CD45 + cells in the blood, spleen, and tumor ( B ) of Il1r2 +/+ ( n = 10) and Il1r2 −/− ( n = 13) FS6-bearing mice. C–F, MFI ( C ) and frequency ( D–F ) of cells positive for selected markers on total neutrophils in the blood, spleen, and tumor of Il1r2 +/+ [ n = 5 ( C ), 10 ( D and E ), and 7 ( F )] and Il1r2 −/− [ n = 8 ( C ), 13 ( D and E ), and 5 ( F )] FS6-bearing mice. G and H, Quantification of ROS production ( G ) and frequency of iNOS + ( H ) cells gated on total neutrophils infiltrating FS6 tumors in Il1r2 +/+ [ n = 10 ( G ) and 21 ( H )] and Il1r2 −/− [ n = 14 ( G ) and 22 ( H )] mice. I and J, MFI ( I ) and frequency ( J ) of cells positive for selected markers on total neutrophils in tumors of Il1r2 +/+ ( n = 7), Il1r2 −/− ( n = 5), Csf3r −/− ( n = 6), and Il1r2 −/− /Csf3r −/− ( n = 4) FS6-bearing mice. K–M, Frequency of CD44 low/high ( K ), PD-1 + ( L ), and CD25 + ( M ) cells on tumor-infiltrating Tregs in Il1r2 +/+ [ n = 14 ( K ) and 16 ( L and M )] and Il1r2 −/− ( n = 12) FS6-bearing mice. Frequency ( N and O ) of cells positive for selected markers on Tregs in the tumor ( N ) and spleen ( O ) of Il1r2 +/+ ( n = 4) and Il1r2 −/− ( n = 5) FS6-bearing mice. Mean ± SEM. Two ( A and B , D and E , G , and K–M ) and four ( H ) pooled experiments; one representative experiment of two ( C ); and one experiment ( F , I , J , N , and O ) performed. P values determined by two-tailed Mann–Whitney U test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Il1r1 −/− (RRID: IMSR_JAX:003245) and Csf3r −/− (RRID: IMSR_JAX:017838) mice were purchased from The Jackson Laboratory.

Techniques: Two Tailed Test, MANN-WHITNEY

Journal: Cancer Immunology Research

Article Title: IL1R2 Deficiency Unleashes Neutrophil-Mediated Antitumor Potential in Sarcoma

doi: 10.1158/2326-6066.CIR-25-0651

Figure Lengend Snippet: Association between IL1R2 deficiency signature, neutrophil infiltration, and survival in patients with UPS. A, Violin plots showing IL1R2 expression levels in myeloid and lymphoid populations from single-cell RNA-seq data (Tabula Sapiens datasets; ref. ) of human donor-derived BM, blood, and spleen (left) and percentage of IL1R2 + in the indicated cell subsets from single-cell RNA-seq data ( GSE212527 dataset) of human sarcomas (right). B, Enrichment score of IL1R2 expression (left) or single-sample GSEA (ssGSEA) enrichment score of the IL1R2 deficiency signature (right) in patients with UPS divided according to their low or high expression of CSF3R or TAN signature. C and D, Scatter plots representing the correlation between CSF3R expression ( C ) or TAN signature ( D ) and IL1R2 deficiency signature enrichment in patients with UPS. Blue line, linear model interpolating curve; darker bands, linear model confidence intervals. E–H, Kaplan–Meier OS curves of total patients with sarcoma ( E and G ) or UPS ( F and H ) from TGCA, divided on the basis of median enrichment score of IL1R2 ( E and F ) or IL1R2 deficiency signature ( n = 180 genes; G and H ). Shaded areas of Kaplan–Meier survival curves represent 95% upper and lower confidence intervals. Enrichment threshold is set based on the median value. I, Enrichment score of the IL1R2 deficiency signature in patients with UPS divided according to their low or high expression of T1, T2, or T3 signature . B–D , F , H , and I, UPS: n = 51 patients; n = 52 tumor samples in total; and n = 26 tumor samples in each group. E and G, Total sarcoma: n = 259 patients; n = 130 patients in the high group; and n = 129 patients in the low group. B and I, Boxes: 25–75 range; whiskers: 10–90 range. Exact P values of the two-sided log-rank test (for survival curve E–H ) and correlation test (Spearman correlation for scatterplots; C and D ). Two-tailed Mann–Whitney U test ( B and I ). ns, not statistically significant; **, P < 0.01; ****, P < 0.0001. ECs, endothelial cells; Mϕ, macrophages; Mono., monocytes; Nϕ, neutrophils; rsem, RNA-sequencing by expectation maximization; SMCs, smooth muscle cells.

Article Snippet: Il1r1 −/− (RRID: IMSR_JAX:003245) and Csf3r −/− (RRID: IMSR_JAX:017838) mice were purchased from The Jackson Laboratory.

Techniques: Expressing, Single Cell, RNA Sequencing, Derivative Assay, Two Tailed Test, MANN-WHITNEY

Journal: Signal Transduction and Targeted Therapy

Article Title: Colony-stimulating factor 3 as a key mediator in the progression of idiopathic pulmonary fibrosis: a novel therapeutic target

doi: 10.1038/s41392-025-02421-6

Figure Lengend Snippet: CSF3 induces pulmonary fibrosis through the CSF3R/STAT3 signaling axis. a qRT-PCR analysis of fibrosis markers (α-SMA, COL1A1, fibronectin) in human lung fibroblast cell line (HLF, left) and mouse primary lung fibroblast cells (MLF, right) treated with recombinant CSF3 (rCSF3) (200 ng/ml, 24 h). b and c Representative immunofluorescence images of α-SMA and COL1A1 ( b ), and Western blot analysis of fibrosis markers and STAT3 activation (p-STAT3) ( c ) in HLF and MLF cells treated with rCSF3. β-Actin was used as a loading control. Scale bars: 200 μm. d Hydroxyproline content assay in HLF and MLF cells treated with rCSF3. e Schematic illustration of the procedure used to induce lung fibrosis in C57BL/6 mice with recombinant mouse CSF3. BLM was administered IP on day 1, followed by IP injections of rCSF3 three times per week. f Quantification of the fibrosis area was performed using Orbit software based on Masson’s trichrome staining images. The green area represents the fibrotic region. Scale bar: 500 μm. g Hydroxyproline content in mouse lung tissues from groups treated with BLM and rmCSF3 as indicated. h–k qRT-PCR analysis of α-SMA ( h ), COL1A1 ( i ) and fibronectin ( j ) expression, and hydroxyproline content ( k ) assay in wild-type (CSF3 +/+ ) MLF cells and CSF3 knock-out (CSF3 −/− ) MLF cells treated with BLM and rCSF3. l and m qRT-PCR analysis of fibrosis markers and CSF3 expression ( l ), and representative immunofluorescence images of α-SMA and COL1A1 ( m ) in HLF cells transfected with si-CSF3R and treated with rCSF3 (200 ng/ml, 24 h). Scale bars: 200 μm. n and o Western blot analysis of α-SMA and COL1A1 ( n ), and representative immunofluorescence images of α-SMA and COL1A1 ( o ) in CSF3 −/− MLF cells treated with BLM and rCSF3. β-Actin was used as a loading control. Scale bars: 200 μm. Statistical significance was determined using ANOVA with multiple comparison or t -test

Article Snippet: The TGFβRII or CSF3R siRNA was purchased from Santa Cruz (Cat. No. sc-36657 for TGFβRII) or Genolution, Inc. for CSF3R.

Techniques: Quantitative RT-PCR, Recombinant, Immunofluorescence, Western Blot, Activation Assay, Control, Software, Staining, Expressing, Knock-Out, Transfection, Comparison