Figure legends
Figure 1. DEP screening and annotation enrichment analysis of the plasma proteome profiles of pediatric LCH patients. (A) Volcano plot of DEPs between the MS LCH and SS LCH groups. The blue points represent downregulated proteins, while the red points represent upregulated proteins. (B) Hierarchical clustering heatmap of the screened DEPs. (C) KEGG annotation and enrichment analysis of DEPs showing the signaling pathway diagram for the enriched items. (D) Gene Ontology annotation and enrichment analysis of DEPs, including biological process (BP), molecular function (MF), and cellular component (CC) terms.
Figure 2. Identification of overlapping DEPs/DEGs by scRNA-seq versus plasma proteomics. (A) T-distributed stochastic neighbor embedding (t-SNE) plot of nine PBMC subclusters from MS RO+ LCH (n= 7) and SS LCH (n=3) patients. (B) Venn diagram illustrating the intersections between the two comparisons according to plasma proteomics versus scRNA-seq. (C) The feature plot shows the clustering of common DEGs (CSF1R , CD14 , and GSN ) based on gene expression. The color legend shows the log10-transformed expression levels of the genes.
Figure 3. sCSF1R levels in plasma and CSF1R expression in LCH lesions. (A) sCSF1R levels in plasma samples from LCH patients and healthy children determined by ELISA. (B) Scatter plots showing the differences in sCSF1R levels among SS, MS RO-, and MS RO+ LCH patients. (C) Representative images of immunofluorescence staining forBRAF- V600E (pink) and CSF1R (green) in LCH skin biopsies. Original magnification: 100×; scale bar: 50 μm. Histogram of MFI (average fluorescence intensity) among SS, MS RO-, and MS RO+ LCH.
Figure 4. Prognostic value of plasma sCSF1R levels at diagnosis. (A) ROC curve schematic. (B) 3-year PFS in the cohort of 104 LCH patients treated with standard first-line treatment. (C) PFS in patients with SS LCH (n= 68). (D) 3-year PFS in patients with MS LCH (n= 36). (E-F) Univariate and multivariate analyses of prognostic factors for PFS in children with LCH.
Figure 5. Monitoring of plasma sCSF1R levels during follow-up in LCH patients. (A) The levels of plasma sCSF1R at two time points (at diagnosis and at week 6) in 23 patients receiving first-line therapy. (B-G) Dynamics of the plasma sCSF1R and cfBRAF -V600E levels in six relapsed patients treated with BRAFi therapy. The black arrows indicate the time points at which patients stopped treatment with darafenib; the red arrows indicate the time of recurrence. Treatment regimens are indicated at the bottom of each graph. The limit of the cfBRAF -V600E detection assay was determined to be 0.1%.