Journal: bioRxiv

Article Title: Inhibiting EZH2 Alleviates Osteoarthritis and Pain in an Experimental Murine Model Through Modulating Synovial and Macrophage Inflammation, Axon Guidance, and Osteoclastogenesis

doi: 10.1101/2025.01.19.633762

Figure Lengend Snippet: Schema of in vivo experimentation (A). Osteoarthritis was induced in mice by intra-articular injection of 0.75 mg of monosodium iodoacetate diluted in 10 μL of sterile saline solution. Two, seven and thirty days after OA induction, EZH2 inhibitor (EPZ-6438, 10 µM, 10 µL) was administrated directly into the right synovial capsule of OA mice. Control mice received injections of vehicle at the same time. At day 56, mice were euthanized and histology were performed. OA knees were stained with Safranin O and counterstained with Fast Green. The slides were observed using a light microscope (B). Cartilage degradation was assessed by the OARSI and Mankin scores (C). Synovial thickness was also measured (D). Results are present as mean ± SEM (n=4). In addition, functional pain was evaluated once a week during all the experiments using Von Frey filaments (E), and at day 56 using static weight-bearing distribution (F). Results are present as median ± interquartile range (n=8). *: p-value < 0.05; **: p-value < 0.01. For figure E, the “vehicle” condition was compared to the “sham” condition (##) or “EPZ-6438” (**).

Article Snippet: The EZH2 inhibitor, EPZ-6438 also called tazemetostat (Selleckchem, Souffelweyersheim, France) was resuspended in dimethylsulfoxide (DMSO, Dutscher, Bernolsheim, France) at a concentration of 10 mM, then diluted directly in medium ( in vitro culture) or in saline solution ( in vivo experiments) to reach the concentration of 10 μM.

Techniques: In Vivo, Injection, Sterility, Saline, Control, Staining, Light Microscopy, Functional Assay

Journal: bioRxiv

Article Title: Inhibiting EZH2 Alleviates Osteoarthritis and Pain in an Experimental Murine Model Through Modulating Synovial and Macrophage Inflammation, Axon Guidance, and Osteoclastogenesis

doi: 10.1101/2025.01.19.633762

Figure Lengend Snippet: Human fibroblast-like synoviocytes were cultured 48h with IL-1β (1 ng/ml). Then, EZH2 mRNA expression was analyzed by RT-PCR (A). Cells stimulated with IL-1β in the presence or not of EPZ-6438 (10 µM) were marked for H3K27me3 (FITC, green) by immunocytochemistry. DAPI was used to stain the nuclei. Representative images of 3 independent experiments were shown. Graph represents the percentage of H3K27me3 positive cells (B). mRNA of IL-1β, MMP-13 and NGF was also determined using RT-PCR (C). Means + SEM of at least 3 independent experiments are shown. *: p-value < 0.05; **: p-value < 0.01; ***: p-value < 0.001. Autophagy was also assessed though the detection of autophagosomes by immunofluorescence (blue signal). Representative images from three independent experiments are shown (D).

Article Snippet: The EZH2 inhibitor, EPZ-6438 also called tazemetostat (Selleckchem, Souffelweyersheim, France) was resuspended in dimethylsulfoxide (DMSO, Dutscher, Bernolsheim, France) at a concentration of 10 mM, then diluted directly in medium ( in vitro culture) or in saline solution ( in vivo experiments) to reach the concentration of 10 μM.

Techniques: Cell Culture, Expressing, Reverse Transcription Polymerase Chain Reaction, Immunocytochemistry, Staining, Immunofluorescence

Journal: bioRxiv

Article Title: Inhibiting EZH2 Alleviates Osteoarthritis and Pain in an Experimental Murine Model Through Modulating Synovial and Macrophage Inflammation, Axon Guidance, and Osteoclastogenesis

doi: 10.1101/2025.01.19.633762

Figure Lengend Snippet: Fibroblast-like synoviocytes were incubated for 48h with IL-1β and the binding of EZH2 on gene promoters were investigated by Chip-Seq. A simple binary comparison (identifying common or unique peaks) was performed, and functional enrichments in Biological Process (Gene Ontology) from genes having unique peaks for EZH2 at their promoters in IL-1β condition were done and visualized using STRING database (A). The network of predicted protein-protein interactions is also showed with blue name corresponding to genes involved in “axon guidance” (B). Peaks obtained from Chip-Seq and corresponding of loci of some of these genes are showed (C).

Article Snippet: The EZH2 inhibitor, EPZ-6438 also called tazemetostat (Selleckchem, Souffelweyersheim, France) was resuspended in dimethylsulfoxide (DMSO, Dutscher, Bernolsheim, France) at a concentration of 10 mM, then diluted directly in medium ( in vitro culture) or in saline solution ( in vivo experiments) to reach the concentration of 10 μM.

Techniques: Incubation, Binding Assay, ChIP-sequencing, Comparison, Functional Assay, Protein-Protein interactions

Journal: Cancers

Article Title: BET and CDK Inhibition Reveal Differences in the Proliferation Control of Sympathetic Ganglion Neuroblasts and Adrenal Chromaffin Cells

doi: 10.3390/cancers14112755

Figure Lengend Snippet: Effect of IGF, EZH2, ALK, and WNT inhibitors on the proliferation of chromaffin cells, neuroblasts, and NESTIN-expressing cells (NECs). Dose–response curves are shown for IGFR inhibitor (PPP) ( A ), EZH2 inhibitor (EPZ6438) ( B ), Alk inhibitor (Alectinib), ( C ) and Wnt inhibitor (ICG001) ( D ). Data represent the mean ± s. e. m. of at least three independent experiments.

Article Snippet: The inhibitors that were used were the BET inhibitors JQ1 (Tocris Biotechne, Wiesbaden, Germany; 4499) and GSK1324726A (iBET 726) (Selleckchem Biozol, Eching, Germany), the CDK-7 inhibitors THZ1 (Medchem Express Biotrend, Köln, Germany) and YKL-5-125 (Selleckchem), the CDK12/13 inhibitor THZ 531 (Selleckchem), the IGF1-R inhibitor picropodophyllin (PPP) (Tocris 2956), the EZH2 inhibitor EPZ 6438 (Axon Medchem, Groningen, NL; 2227), the WNT inhibitor ICG001 (Axon Medchem 1766), and the ALK inhibitor Alectinib (Selleckchem S276).

Techniques: Expressing