human myeloma cell lines hmcls jjn 3  (DSMZ)

Journal: bioRxiv

Article Title: The KDM6 histone demethylase inhibitor GSK-J4 induces metal and stress responses in multiple myeloma cells

doi: 10.1101/2024.12.28.630531

Figure Lengend Snippet: (A) Heatmap reporting as % change over solvent control (DMSO) in the viability of multiple myeloma cell lines (AMO1, JJN3, RPMI8226, MM1S), screened against a focused library of epigenetic tool compounds (SI Data 1) as measured using PrestoBlue assay. (B) Structures of GSK-J4 and analogues used in this study. (C) Binding mode and chelation of active site metal in human KDM6B by GSK-J1 (presented as sticks, PDB code 4ASK). Sphere depicts position of the Co2+ metal used in the crystallisation experiments, amino acid residues of KDM6B (His1390, Glu1392) involved in metal coordination are depicted. (D) EC 50 values for anti-proliferative effects measured in multiple myeloma cell lines treated with GSK-J4 or GSK-J5. (E) Dose response curve for GSK-J4 in JJN3 cells (EC 50 = 1.1 μM). (F) Flow cytometry data for JJN3 cells and induction of cell death upon 72 hours of DMSO, GSK-J4 and GSK-J5 treatment (2 μM).

Article Snippet: AMO-1 and JJN3 cells lines were purchased from DSMZ (Braunschweig, Germany) or LGC standards (Teddington, UK).

Techniques: Solvent, Control, Prestoblue Assay, Analogues, Binding Assay, Flow Cytometry

Journal: bioRxiv

Article Title: The KDM6 histone demethylase inhibitor GSK-J4 induces metal and stress responses in multiple myeloma cells

doi: 10.1101/2024.12.28.630531

Figure Lengend Snippet: (A) A heatmap displaying the differentially regulated genes identified by microarray analysis in JJN3 and RPMI cell lines treated with various concentrations of DMSO (0.1%) or GSK-J4 (2 μM and 0.5 μM) concentration. The map highlights the known associations with the transcription factors ATF4 or MTF1, and includes a contrast with JJN3 cells treated with the proteosome inhibitor Bortezomib (10 nM) (B-D). Volcano plots showing selected differential gene expression responses from RNA-seq data for (B) 3 hrs, (C) 6 hrs and (D) 24 hrs following GSK-J4 treatment (2 μM). (E) Motif enrichment analysis showing the top two enriched motifs at the 24 hr timepoint. (F) GO analysis showing the top enriched ontology terms at the 24 hr timepoint. (G) Scatter plots showing the expression of MT1X and MT1G following treatment of JJN3 cells with either 3 hrs, 6 hrs or 24 hrs of DMSO or GSK-J4 treatment (2 μM). (H) RT-PCR analysis of MT1X gene expression of JJN3 cells treated with GSK-J4 for 1, 3, 6, 12 and 24 hrs. (I) UMAP of single-cell sequencing of human bone marrow samples (newly diagnosed myeloma), depicting the major cell type clusters identified. The top right panel shows the expression of MT1X treated with either DMSO (laft bar) or GSK-J4 (right bar; 2 μM). The bottom right panel shows the expression of MT1G treated with either DMSO (left bar) or GSK-J4 (right bar; 2 μM).

Article Snippet: AMO-1 and JJN3 cells lines were purchased from DSMZ (Braunschweig, Germany) or LGC standards (Teddington, UK).

Techniques: Microarray, Concentration Assay, Expressing, RNA Sequencing Assay, Reverse Transcription Polymerase Chain Reaction, Sequencing

Journal: bioRxiv

Article Title: The KDM6 histone demethylase inhibitor GSK-J4 induces metal and stress responses in multiple myeloma cells

doi: 10.1101/2024.12.28.630531

Figure Lengend Snippet: (A) Left panel shows representative images of FluoZin-3 stained JJN3 cells at baseline, 48 mins and 120 mins following GSK-J4 treatment (2 μM). The right panel shows the relative fluorescence of FluoZin-3 in 3 representative cells over 120 mins of exposure with GSK-J4. (B) Dose response curve for ZnSO 4 treatment in JJN3 cells. (C) Dose response curve for GSK-J4 treatment alone or co-treatment of GSK-J4 and ZnSO 4 in JJN3 cells.

Article Snippet: AMO-1 and JJN3 cells lines were purchased from DSMZ (Braunschweig, Germany) or LGC standards (Teddington, UK).

Techniques: Staining, Fluorescence

Journal: bioRxiv

Article Title: The KDM6 histone demethylase inhibitor GSK-J4 induces metal and stress responses in multiple myeloma cells

doi: 10.1101/2024.12.28.630531

Figure Lengend Snippet: (A) Heatmap of all differentially expressed genes in JJN3 cells treated with either DMSO (0.1%), GSK-J4 (2 μM) or ZnSO 4 (100 μM) after 1, 3, 6 and 24 hrs. (B) Venn diagram showing the overlap of all differentially regulated genes across all timepoints following treatment with GSK-J4 and ZnSO 4 . (C) STRING analysis of genes overlapping between GSK-J4 and ZnSO 4 . (D) GO analysis of showing the ontologies that are enriched for overlapping genes between GSK-J4 and ZnSO 4 . (E-F) Heatmaps showing the expression of selected stress response (E) or transcription and chromatin factor (F) genes following GSK-J4 or ZnSO 4 treatment for 1, 3, 6 and 24 hrs.

Article Snippet: AMO-1 and JJN3 cells lines were purchased from DSMZ (Braunschweig, Germany) or LGC standards (Teddington, UK).

Techniques: Expressing

Journal: Biomolecules

Article Title: Preclinical Evaluation of a Novel Series of Polyfluorinated Thalidomide Analogs in Drug-Resistant Multiple Myeloma

doi: 10.3390/biom14060725

Figure Lengend Snippet: Treatment with Gu compounds inhibited multiple myeloma cell proliferation in vitro. ( A ) The RPMI-8226 cell proliferation during 72-h treatment was most potently inhibited by Gu1210, Gu1213, Gu1214, and Gu1215 at a concentration of 10 μM compared to that of the vehicle control (0.5% DMSO). ( B ) The JJN3 cell proliferation during 24-h treatment was most potently inhibited by Gu1210, Gu1213, Gu1214, and Gu1215 at a concentration of 10 μM compared to that of the vehicle control (0.5% DMSO). ( C ) The 72-h treatment of RPMI-8226 cells with four lead analogs at a range of concentrations (0.1 μM–20 μM) showed a dose-dependent response, with Gu1215 exhibiting the most potent inhibition (**** p < 0.0001). ( D ) The 24-h treatment of JJN3 cells with the four lead analogs at a range of concentrations (0.1 μM–20 μM) showed a dose-dependent response, with Gu1215 exhibiting the most potent inhibition (**** p < 0.0001). Note: a 5 µM dose represents two independent replicates. ( E ) The 72-h treatment of MOLP-8 cells with the four lead analogs at a range of concentrations (0.1 μM–10 μM) showed dose-dependent responses, with Gu1214 exhibiting the most potent inhibition (**** p < 0.0001). These data are representative of at least three independent experiments with at least three replicates per experiment. In all the figures, any cases in which the concentrations are grouped in a bracket indicate that all of the values within the bracket are individually statistically different from the vehicle control.

Article Snippet: The human myeloma cell line JJN3 (RRID:CVCL_2078) was purchased from DSMZ (Leibniz Institute DSMZ–German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) and grown in 40% DMEM and 40% IMDM (Life Technologies, Carlsbad, CA, USA) with 20% FBS.

Techniques: In Vitro, Concentration Assay, Control, Inhibition