lkb1 specific inhibitor pim1 akk1  (MedChemExpress)

Journal: Inflammation

Article Title: Keratinocyte-derived Exosomal LKB1 Drives Vitiligo Progression by Activating CD8 + T Cells

doi: 10.1007/s10753-026-02479-6

Figure Lengend Snippet: Exosomal LKB1 serves as a key mediator of vitiligo progression. Proteomic profiling of differentially expressed proteins between Vexo versus Hexo ( A ) and H₂O₂/exo versus Con/exo ( B ), screened using p < 0.05 and fold change > 2. Red indicates upregulated proteins; blue indicates downregulated proteins. ( C ) Venn diagram showing the intersection of proteins upregulated in both Vexo versus Hexo and H₂O₂/exo versus Con/exo datasets, identifying LKB1 as the sole common candidate. ( D ) Western blot analysis of LKB1 expression in keratinocytes isolated from lesional skin of three vitiligo patients and keratinocytes from three healthy individuals. ( E ) Multiplex immunofluorescence staining of CD8 (red) and CD69 (green) expression in lesional skin from vitiligo patients with nuclear counterstaining by DAPI (blue). ( F ) Multiplex immunofluorescence staining of LKB1 (red) and CD69 (yellow) expression in lesional skin from vitiligo patients and healthy control skin, with nuclear counterstaining by DAPI (blue). ( G ) Quantification of LKB1 expression in skin biopsies from 12 vitiligo patients and 12 healthy controls. ( H ) Correlation analysis between LKB1 expression and CD69 expression in the lesional skin of vitiligo patients. Vexo, exosomes derived from keratinocytes of vitiligo patients; Hexo, exosomes derived from healthy donors; H 2 O 2 /exo, exosomes derived from H 2 O 2 -treated HaCaT cells; Con/exo, exosomes derived from untreated HaCaT cells; * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant

Article Snippet: Vitiligo mice treated with 40 mg/kg LKB1 inhibitor (HY-10371, MCE, USA) administered intraperitoneally.

Techniques: Western Blot, Expressing, Isolation, Multiplex Assay, Immunofluorescence, Staining, Control, Derivative Assay

Journal: Inflammation

Article Title: Keratinocyte-derived Exosomal LKB1 Drives Vitiligo Progression by Activating CD8 + T Cells

doi: 10.1007/s10753-026-02479-6

Figure Lengend Snippet: Exosomal LKB1 drives CD8⁺ T cells activation and vitiligo progression. Western blot analysis of LKB1 expression in CD8⁺ T cells treated with Hexo or Vexo ( A ) and Con/exo or H₂O₂/exo ( B ). ELISA quantification of granzyme B ( C ) and IFN-γ ( D ) secretion by CD8⁺ T cells treated with exosomes from LKB1-overexpressing HaCaT cells (HaCaT/exo LKB1 ) or vector control exosomes (HaCaT/exo vector ). ELISA quantification of granzyme B ( E ) and IFN-γ ( F ) secretion by CD8⁺ T cells treated with exosomes from H₂O₂-treated HaCaT cells with LKB1 knockdown (H₂O₂/exo shLKB1 ) or negative control shRNA (H₂O₂/exo shNC ). Flow cytometric analysis of melanocyte apoptosis following co-culture with CD8⁺ T cells treated with HaCaT/exo LKB1 or HaCaT/exo vector , shown as representative plots ( G ) and quantitative summary ( H ). ( I ) Representative tail skin photographs of vitiligo mice from the untreated control group and the group treated with 40 mg/kg LKB1 inhibitor administered intraperitoneally. Immunofluorescence staining of CD69 expression in tail skin from control and LKB1 inhibitor–treated vitiligo mice, shown as representative images ( J ) and quantification ( K ). Vexo, exosomes derived from keratinocytes of vitiligo patients; Hexo, exosomes derived from healthy donors; HaCat/exoLKB1, exosomes derived from LKB1-overexpressing HaCat keratinocytes; H 2 O 2 /exo shLKB1 , exosomes derived from LKB1-knockdown keratinocytes under H 2 O 2 treat; * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant

Article Snippet: Vitiligo mice treated with 40 mg/kg LKB1 inhibitor (HY-10371, MCE, USA) administered intraperitoneally.

Techniques: Activation Assay, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Control, Knockdown, Negative Control, shRNA, Co-Culture Assay, Immunofluorescence, Staining, Derivative Assay

Journal: Inflammation

Article Title: Keratinocyte-derived Exosomal LKB1 Drives Vitiligo Progression by Activating CD8 + T Cells

doi: 10.1007/s10753-026-02479-6

Figure Lengend Snippet: Exosomal LKB1 mediates CD8 + T cells activation via the AMPK pathway. ( A ) Heatmap of RNA-seq analysis showing differentially expressed genes in CD8⁺ T cells treated with Hexo or Vexo. ( B ) KEGG pathway enrichment analysis of differentially expressed genes in Vexo-treated versus Hexo-treated CD8⁺ T cells, highlighting significant activation of the AMPK signaling pathway. ( C–D ) Western blot analysis of LKB1 and p-AMPK expression in CD8⁺ T cells treated with different exosome preparations (Hexo, Vexo, H₂O₂/exo, or Con/exo) in the presence or absence of LKB1 inhibitor (10 µM). ( E ) Western blot analysis of p-AMPK expression in CD8⁺ T cells after LKB1 overexpression. ELISA quantification of granzyme B ( F ) and IFN-γ ( G ) secretion by CD8⁺ T cells treated with indicated exosome groups with or without AMPK inhibition (20 µM). ( H ) Flow cytometric analysis of IFN-γ⁺ CD8⁺ T cells following treatment with indicated exosome preparations. Vexo, exosomes derived from keratinocytes of vitiligo patients; Hexo, exosomes derived from healthy donors; LBK1in, LKB1 inhibitor; H 2 O 2 /exo, exosomes derived from H 2 O 2 -treated HaCaT cells; Con/exo, exosomes derived from untreated HaCaT cells; LKB1-oe, LBK1 overexpression; * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant

Article Snippet: Vitiligo mice treated with 40 mg/kg LKB1 inhibitor (HY-10371, MCE, USA) administered intraperitoneally.

Techniques: Activation Assay, RNA Sequencing, Western Blot, Expressing, Over Expression, Enzyme-linked Immunosorbent Assay, Inhibition, Derivative Assay

Journal: Inflammation

Article Title: Keratinocyte-derived Exosomal LKB1 Drives Vitiligo Progression by Activating CD8 + T Cells

doi: 10.1007/s10753-026-02479-6

Figure Lengend Snippet: IFN-γ secreted by CD8 + T cells positively feeds back on the excessive production of LKB1 in keratinocytes. ( A ) qRT-PCR analysis of LKB1 expression in HaCaT cells at various time points following IFN-γ treatment. qRT-PCR ( B ) and Western blot ( C ) analyses of LKB1 expression in HaCaT cells treated with increasing concentrations of IFN-γ. ( D ) Schematic representation of the co-culture system between keratinocytes and CD8⁺ T cells pretreated with Hexo or Vexo. qRT-PCR analysis of LKB1 expression in HaCaT cells co-cultured with CD8⁺ T cells treated with Vexo ( E ) or H₂O₂/exo ( F ). ( G ) Western blot analysis of p-STAT1 levels in HaCaT cells following IFN-γ stimulation. ( H ) Chromatin immunoprecipitation–qRT-PCR analysis showing recruitment of STAT1 to the LKB1 promoter region. ( I ) Predicted STAT1-binding motifs within the LKB1 promoter identified from the JASPAR database. A dual luciferase reporter plasmid containing the wild-type or mutant binding sites (#1–3) was constructed. ( J ) Luciferase activity in HEK293T cells co-transfected with the indicated LKB1 promoter constructs and STAT1 expression plasmid. qRT-PCR ( K ) and Western blot ( L ) analyses of LKB1 expression in HaCaT cells treated with IFN-γ in the presence or absence of a STAT1 inhibitor (5 µM). * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant

Article Snippet: Vitiligo mice treated with 40 mg/kg LKB1 inhibitor (HY-10371, MCE, USA) administered intraperitoneally.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Co-Culture Assay, Cell Culture, Chromatin Immunoprecipitation, Binding Assay, Luciferase, Plasmid Preparation, Mutagenesis, Construct, Activity Assay, Transfection

Journal: Inflammation

Article Title: Keratinocyte-derived Exosomal LKB1 Drives Vitiligo Progression by Activating CD8 + T Cells

doi: 10.1007/s10753-026-02479-6

Figure Lengend Snippet: Proposed mechanistic model of keratinocyte-derived exosomal LKB1 in vitiligo pathogenesis. Schematic illustration of the proposed pathogenic feedback loop in vitiligo. Vitiligo lesional keratinocytes secrete exosomes enriched in LKB1 (Vexo), which are internalized by CD8⁺ T cells. Exosomal LKB1 activates the AMPK signaling pathway in CD8⁺ T cells, leading to increased secretion of IFN-γ and granzyme B, thereby enhancing melanocyte-directed cytotoxicity. IFN-γ released by activated CD8⁺ T cells further stimulates keratinocytes via the JAK/STAT1 pathway, resulting in increased LKB1 expression and further exosome release. This establishes a self-amplifying loop between keratinocytes and CD8⁺ T cells that sustains autoimmune melanocyte destruction and promotes vitiligo progression

Article Snippet: Vitiligo mice treated with 40 mg/kg LKB1 inhibitor (HY-10371, MCE, USA) administered intraperitoneally.

Techniques: Derivative Assay, Expressing