Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Psoriasis evokes pain‐associated coping behaviors. (A) Representative images of dorsal skin and surrounding hair in Vas‐ and IMQ‐treated mice. Red‐boxed areas are magnified to highlight changes in fur around lesions. (B) Quantification of licking events over 1 h. (C) Representative immunofluorescence images of β3‐tubulin in healthy controls (HC) and psoriasis patients (Ps). (D) The %β3‐tubulin‐positive staining area in C. (E) Representative movement tracks of mice in the open field arena during a 10‐min session. (F) Total distance traveled (m) in the open field. (G) Number of entries into the central zone. (H) Duration spent in the central zone (s). (I) Diagram of the IL‐17A‐induced psoriasis model in C57BL/6 mice injected in hind paws with PBS or IL‐17A (created using Figdraw). (J) H&E staining of PBS‐ and IL‐17A‐injected hind paw skin. (K) Quantification of epidermal thickness in PBS‐ versus IL‐17A‐injected skin. (L) Paw withdrawal threshold (g) measured using Von Frey filaments. (M) Latency to jumping (s) on a hot plate (55°C). (N) Number of hind paw lifts during 5‐min cold plate exposure (4°C). (O) Total duration of hind paw lifting (s) during cold plate exposure. Scale bars as indicated. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; according to unpaired Student's t ‐test (B, D, F, G, H, K, L, M, N, O). n = 3 (D), n = 6 (B, F, G, H, K, L, M, N, O).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Immunofluorescence, Staining, Injection

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Psoriatic lesions exhibit elevated NGF with cutaneous TrkA loss and DRG accumulation. (A) Representative fluorescent immunohistochemistry images and quantification of NGF mean fluorescence intensity of NGF in healthy controls (HC) and psoriasis patients (Ps). (B) Quantification of NGF levels in skin tissues from healthy controls (HC) and psoriasis patients. (C) Western blot analysis of NGF and IL‐17A expression in dorsal skin of Vas‐ and IMQ‐treated mice. (D) Statistical analysis of NGF/IL‐17A expression from C. (E) Western blot analysis of p‐TrkA and TrkA protein expression in dorsal skin of Vas‐ and IMQ‐treated mice. (F) Quantification of p‐TrkA and TrkA expression from E. (G) Schematic of neurons with cell bodies in the dorsal root ganglia (DRG) projecting axons to the skin. (H) Western blot analysis of protein expression in DRG of Vas‐ and IMQ‐treated mice. (I) Quantification of protein expression from H. (J) Relative c‐Fos mRNA expression in DRG from Vas‐ and IMQ‐treated mice. In (E), the two arrow‐indicated bands together represent the total TrkA signal in mouse skin and were quantified in (F). Scale bar as indicated. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001; unpaired Student's t ‐test (A, B, D, F, I, J); n = 3 (A, B, I, J), n = 6 (D, F).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Immunohistochemistry, Fluorescence, Western Blot, Expressing

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: IL‐17A and NGF are associated with MLC2‐Linked cytoskeletal remodeling in Schwann cells. (A) Representative TEM images of myelinated bundles in the dorsal skin of Vas‐ and IMQ‐treated mice. (B) TEM images of myelinated bundles in PBS‐ or IL‐17A‐injected hind paws. (C) TEM images of glio‐neural complexes in PBS‐ or IL‐17A‐injected hind paws. (D) TEM images of glio‐neural complexes in healthy controls (HC) and psoriasis patients (Ps). Red arrows indicate Schwann cell nuclei in (A, B). Glio‐neural complexes at the dermal–epidermal junction were pseudo‐colored: axons (green), Schwann cells and processes (red) in (C, D). (E) Representative fluorescence images of Schwann cells under PBS (control) or IL‐17A/NGF treatment, stained with phalloidin. (F–H) Quantification of Schwann cell morphology index (F), relative cell area (G), and F‐actin mean intensity (H) under control conditions. (I, J) Schwann cells were stimulated with NGF (100 ng/mL, I) or IL‐17A (20 ng/mL, J) at the indicated time points, and total MLC2 and p‐MLC2 levels were analyzed by Western blot. Scale bars as indicated. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; one‐way ANOVA with post hoc Tukey (F–H), n = 5 (F–H), n = 3 (I, J).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Injection, Fluorescence, Control, Staining, Western Blot

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Schwann cell proliferation in psoriatic‐like lesions. (A) Representative immunofluorescence images of mouse skin stained for SOX10 (Schwann cells, red), β3‐Tubulin (nerve fibers, green), and DAPI (nuclei, blue). (B) Western blot analysis of Schwann cell‐ and nerve‐related proteins in Vas‐ and IMQ‐treated mice. (C) Quantification of S100B, SOX10, MBP, and β3‐Tubulin from B. (D) Workflow schematic for isolation of primary Schwann cells from skin. (E) Representative flow cytometry plot showing identification of Schwann cells gated as SOX10 + S100B + . (F) Quantification of SOX10 + S100B + cells as a percentage of live singlets. (G) Representative immunofluorescence images of Ki67+ RSC96 cells (green) after IL‐17A stimulation (20 ng/mL); nuclei labeled with DAPI (blue). (H) Quantification of Ki67+ RSC96 cells. Scale bars as indicated. * p < 0.05, *** p < 0.001, **** p < 0.0001; unpaired Student's t ‐test (C, F, H); n = 6 (C), n = 3 (F, H).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Immunofluorescence, Staining, Western Blot, Isolation, Flow Cytometry, Labeling

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Schwann cells attenuate NGF‐Driven DRG TrkA signaling via p75NTR, while IL‐17 potentiates the response. (A) PCR and agarose gel electrophoresis analysis of p75NTR and TrkA in Schwann cells (SCs). (B, C) Validation of p75NTR knockdown in primary skin SCs and SCs using siRNA. (D, E) Western blot analysis of p75NTR expression in primary skin SCs and SCs treated with increasing NGF concentrations; accompanying quantification is shown. (F) Western blot and quantification of p75NTR in SCs treated with CHX (70 µg/mL) for the indicated times, with or without NGF (12 h). (G) Schematic of the experimental design for SCs, PC12 cells, and co‐culture under ±NGF and ±IL‐17A conditions (created using Figdraw). (H) Western blot of p‐TrkA and total TrkA in indicated cells after 5‐min NGF (100 ng/mL) stimulation; quantification of p‐TrkA/TrkA ratio shows reduced TrkA phosphorylation in the co‐culture relative to PC12 cells alone. (I) Immunofluorescence of p‐TrkA (red), GFP (green; lentiviral marker in PC12), and DAPI (blue); arrowheads indicate GFP‐negative/DAPI‐positive nuclei corresponding to SCs in the co‐culture. (J) Western blot and quantification of TrkA phosphorylation in co‐cultures treated with NGF and IL‐17A. (K) Schematic of DRG neuron and RSC96 co‐culture assay ±IL‐17A; DRG neurons cultured independently served as controls (created using Figdraw). (L) Representative immunofluorescence images of neurite outgrowth in co‐culture. (M) Quantification of average neurite area per cell. Scale bars as indicated. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; one‐way ANOVA with post hoc Tukey (D, E), two‐way ANOVA with post hoc Tukey (F, M), unpaired Student's t ‐tests (H, J), n = 3 (D, E, F, H, J), n = 17 (M).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Agarose Gel Electrophoresis, Biomarker Discovery, Knockdown, Western Blot, Expressing, Co-Culture Assay, Phospho-proteomics, Immunofluorescence, Marker, Co-culture Assay, Cell Culture

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: IL‐17A enhances NGF secretion and p75NTR regulation in Schwann cells. (A, B) Western blot and quantification of p75NTR and NGF in SCs stimulated with IL‐17A at the indicated time points. (C, D) Dot plots showing the expression of p75NTR and Ngf in SCs from the Vas and IMQ groups. Dot size indicates the percentage of cells expressing each gene, and dot color represents the average expression level. (E) Changes in p75NTR protein levels in SCs treated with IL‐17A (20 ng/mL) in the presence of bafilomycin A1 (Baf A1, 40 nM) or MG132 (20 µM). (F) Western blot and quantification of NGF expression in RSC96 cells treated with IL‐17A for the indicated times. (G) Western blot of NGF in supernatants of RSC96 cells stimulated with IL‐17A (20 ng/mL, 12 h); GM130 served as a negative control to confirm the absence of cellular contamination. (H) Schematic of PC12 treatment with RSC96‐derived conditioned media stimulated by IL‐17A (created using Figdraw). (I) Western blot of PC12 lysates after 5‐min exposure to RSC96‐conditioned media. (J) Quantification of protein levels in I. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; one‐way ANOVA with post hoc Tukey (B, E, F, J), unpaired Student's t ‐test (G), n = 3.

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Western Blot, Expressing, Negative Control, Derivative Assay

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Keap1 downregulation mediates IL‐17A‐Induced Nrf2 activation and NGF upregulation. (A) mRNA expression of Nrf2 and Keap1 in RSC96 cells treated with IL‐17A (20 ng/mL) at indicated times. (B) Western blot analysis of Nrf2 and Keap1 protein levels in RSC96 cells under IL‐17A (20 ng/mL) treatment. (C) Quantification of protein expression from B. (D) Representative immunofluorescence images of Nrf2 (green) in RSC96 cells. (E) Western blot of nuclear and cytosolic Nrf2 in RSC96; Lamin B and GAPDH served as nuclear and cytoplasmic loading controls. (F) Quantification of nuclear Nrf2 from E. (G) Western blot of RSC96 lysates treated with IL‐17A and ML385 (5 µM). (H) Quantification of protein levels in G. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; unpaired Student's t ‐test (F), one‐way ANOVA with post hoc Tukey (A, C, H), n = 3.

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Activation Assay, Expressing, Western Blot, Immunofluorescence

Journal: Advanced Science

Article Title: Depletion of p75NTR in Schwann Cells Driven by Inflammation Mediates Cutaneous Pain in Psoriasis

doi: 10.1002/advs.202523189

Figure Lengend Snippet: Schwann cell–targeted p75NTR overexpression attenuates IL‐17A–evoked nociceptive sensitization. (A) Schematic of the AAV expression cassette. (B) Experimental timeline: plantar AAV injection on day 1; in vivo imaging validation on day 21; repeated plantar IL‐17A (or PBS) injections on days 21, 23, 25, and 27; followed by behavioral testing. (C) Representative in vivo fluorescence imaging showing localized reporter signals at the hind‐paw injection sites in AAV‐Vector and AAV‐p75NTR mice. (D) Immunoblotting of p75NTR in mouse plantar skin lysates with GAPDH as a loading control, and quantification of relative p75NTR protein levels. (E) Mechanical sensitivity assessed by von Frey test (withdrawal threshold, g) in four groups. (F,G) Cold plate test showing the number of paw‐lifting events (F) and lifting duration (G). (H) Hot plate test showing thermal latency. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; unpaired Student's t ‐test (D), one‐way ANOVA with post hoc Tukey (E–H), n = 4 (D), n = 8 (E‐H).

Article Snippet: Human IL‐17A protein (HY‐ P70527 , MCE), Rat IL‐17A protein (HY‐ P78556 , MCE), human β‐NGF (HZ‐1222, Proteintech), Tanezumab (HY‐P99221, MCE), ML385 (HY‐100523, MCE) and CPZ (HY‐12708, MCE) were used for cell stimulation as specified in the experiments. siRNAs targeting human p75NTR ( NGFR ) and a negative control siNC were synthesized commercially.

Techniques: Over Expression, Expressing, Injection, In Vivo Imaging, Biomarker Discovery, In Vivo, Fluorescence, Imaging, Plasmid Preparation, Western Blot, Control, Hot Plate Test