Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Anti‐GBM disease causes tubular damage and partial renal resistance to FGF23. (A) Depicts the experimental workflow in male C57BL/6 mice undergoing induction of anti‐GBM disease using nephrotoxic serum followed by 6 days of intravenous (IV) FGF23 or vehicle treatment. (B) Shows the glomerular filtration rate of different experimental groups at days 0 and 7. (C) Shows urinary albumin/creatinine ratio at day 7. (D) Shows the example renal sections negative or positive for renal tubular casts (arrows) and quantitative tubular cast scores. (E) Shows plasma phosphate and fractional excretion of phosphate of healthy mice and mice with anti‐GBM treated with vehicle or FGF23. Analyses in panel (B): paired t ‐test. Analyses in panels (C–E): two‐way ANOVA. Anti‐GBM, anti‐glomerular basement membrane; d, disease state; t, treatment.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Filtration, Clinical Proteomics, Membrane

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Six‐day course of FGF23 treatment induces renal transcriptional signatures of inflammatory responses and injury. (A) Indicates the number of differentially expressed genes according to experimental comparison in renal bulk RNA‐Seq. (B, C) Depict volcano plots of DEGs above a cutoff of adjusted p < 0.05 and log2‐fold change >1, in the comparison of FGF23 versus vehicle effect in mice with anti‐GBM (B) and the interaction between treatment and disease effect (C). (D–F) Depict significant Reactome gene set enrichment analyses of FGF23 effects in healthy mice (D), mice with anti‐GBM disease (E), and the interaction between treatment and disease effect (F). (G) Depicts a network of ligand–receptor interaction pairs that were significant for FGF23 versus vehicle comparisons in mice with anti‐GBM disease by bulk RNA‐Seq. The ligand–receptor interactions were inferred using R/BulkSignalR. Anti‐GBM, anti‐glomerular basement membrane disease. N = 3 for anti‐GBM groups and n = 4 for healthy groups.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Comparison, RNA Sequencing, Membrane

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Bulk RNA‐Seq deconvolution and immunofluorescence staining reveal an FGF23‐driven increase in overall immune cell and macrophage abundance in the kidneys of mice with anti‐GBM disease. (A, B) Depict the annotation of renal cell clusters in the reanalysis of the single‐cell RNA‐Seq dataset GSE107585 of murine kidney from 7 sex‐mixed healthy C57BL/6 mice; see also Supporting Information Figure . (C) Shows a wedding pie plot of the bulk deconvolution of the renal cellular composition according to FGF23 treatment and anti‐GBM disease state, as indicated by labels. Overall renal immune cells and macrophage‐like cells are displayed by bulk deconvolution (D, G) and by immunofluorescence with automated quantification for CD45 (E, F) and F4/80 (H, I). RNA‐Seq: N = 3 for anti‐GBM groups and n = 4 for healthy groups. Immunofluorescence: n = 4 per group. Statistical analysis: two‐way ANOVA. anti‐GBM, anti‐glomerular basement membrane disease; Baso, basophil; CD, collecting duct; d, disease state; DCT, distal convoluted tubule; DLOH, descending limb of Henle; EC, endothelial cell; Granul, granulocyte; i, interaction; IC, intercalated cells; Ly, lymphocyte; Mono, monocyte; Mph, macrophage; NK, natural killer cell; PC, principal cells; PT, proximal tubule; S, segment; t, treatment.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: RNA Sequencing, Immunofluorescence, Staining, Single Cell, Membrane

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Immune protein profiling highlights an increase in circulating soluble tumor necrosis factor receptors induced by FGF23 and anti‐GBM disease in mice. A plasma cytokine protein array shows FGF23 effects in healthy male C57BL/6 mice (A) and in mice treated with nephrotoxic serum to induce anti‐GBM disease (B). The interaction between treatment and disease state (C) and the overall disease effect (D) are shown. (E–F) Depict analyses of soluble TNF receptors 1 and 2 by two‐way ANOVA. d, disease state; t, treatment. N = 4 biologically independent replicates per group. Anti‐GBM, anti‐glomerular basement membrane.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Clinical Proteomics, Protein Array, Membrane

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Renal immune cell recruitment driven by FGF23 excess is exposure time dependent. Renal microarray transcriptome datasets GDS3361 of male Fgf23 transgenic and control mice or sex‐matched Hyp mice of dataset GDS879 (B) underwent bulk deconvolution with reference to single‐cell RNA‐Seq dataset GSE107585 of murine kidney from 7 sex‐mixed healthy C57BL/6 mice and, subsequently, visualization of overall fractions of inferred immune cells and macrophage‐like cells, as indicated. (C) Shows the experimental workflow of experiments with female BALB/c mice undergoing Adriamycin (doxorubicin) nephropathy followed by a single intravenous (IV) injection of FGF23 or vehicle. (D) Shows the urinary albumin/creatinine ratio 7 days after induction of Adriamycin nephropathy. Statistical analysis: two‐way ANOVA. d, disease state. (E–G) Show significant Reactome gene set enrichment analyses of renal FGF23 effects in healthy mice (E), mice with Adriamycin nephropathy (F), and the interaction between treatment and disease effect (G). (H) Depicts the log‐fold change of 8 transcripts with lowest adjusted p ‐value in the interaction analysis of FGF23 effect in diseased versus FGF23 effect in healthy mice in a 2 × 2 factorial design. N = 5 (A), 10 (B), or 4 (C–H) biologically independent replicates per group.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Microarray, Transgenic Assay, Control, Single Cell, RNA Sequencing, IV Injection

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: Intrarenal proinflammatory effects of FGF23 applied ex vivo in PCKS. (A) Male DBA/2J mice underwent dietary treatment with 0.2% adenine for 15 weeks, followed by organ collection and preparation of 300 µm PCKS for a 24‐h treatment with FGF23 or vehicle ex vivo. No disease‐free controls were used for this substudy. (B) Depicts the fibrotic changes in a representative 4 µm section of PCKS stained with hematoxylin and eosin (left scale bar, 500 µm; right, 100 µm). (C) Shows the log‐fold change of upregulated transcripts with the lowest adjusted p ‐value in the FGF23 versus vehicle comparison. (D–F) Show gene set enrichment analyses of FGF23 effects in Reactome (D), WikiPathways (E), and Pathway Interaction Database (F) gene sets. N = 4 biologically independent replicates per group. PCKS, precision‐cut kidney slices.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: Ex Vivo, Staining, Comparison

Journal: Journal of Cell Communication and Signaling

Article Title: The renal response to FGF23 shifts from phosphaturia toward inflammation in kidney disease

doi: 10.1002/ccs3.70061

Figure Lengend Snippet: FGF23 is associated with renal immune cell content in human patients with IgA nephropathy. As a reference, single‐nucleus RNA‐Seq data from 5 human kidney biopsies ( GSE199711 ) of 2 healthy controls and 3 patients with chronic kidney disease (CKD) were reanalyzed and annotated (A); see also Supporting Information Figure . 35 patients with IgA nephropathy from the Karolinska Kidney Biopsy Cohort showed an inverse univariable association between circulating FGF23 and measured glomerular filtration rate (GFR) (B). The associations between transcriptome‐inferred renal fibroblasts and immune cells (C–D) or macrophages (E–F) and circulating FGF23 are shown, with adjustment for GFR, 25OH‐vitamin D and parathyroid hormone. (D, F) Show disaggregated data stratified by GFR in CKD stages I–II and III–V.

Article Snippet: Recombinant human FGF23 was obtained from R&D Systems, Minneapolis, MN, USA, and distributed via Thermo Fisher (Cat. #100‐52).

Techniques: RNA Sequencing, Filtration