human chronic myelogenous leukemia cell line k562  (ATCC)

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Anti-myeloma activity of the CXCR4 antagonist WZ811

doi: 10.1007/s00109-026-02650-4

Figure Lengend Snippet: CXCR4 expression levels in MM. A The expression of CXCR4 at the protein level was analyzed in a panel of 14 MM cell lines (MM.1S, OPM-1, OPM-2, RPMI-S, RPMI-DOX6, RPMI-DOX40, RPMI-LR5, RPMI-MR20, JJN-3, KMS-11, L-363, OCI-My5, OCI-My7, and U266) by flow cytometry (FACS Canto II flow cytometer, Becton Dickinson), represented by histograms of CXCR4-BV421 positive expression (blue) compared to the negative Ig isotype control (grey), and B by western immunoblot analysis using anti-CXCR4 and anti-GAPDH (used as a loading control) antibodies. The fold change of the geometric mean of CXCR4 expression relative to the isotype fluorescent control is shown. The data are representative from three independent experiments. C CXCR4 expression at the transcriptional mRNA level was examined using RT-PCR, showing mRNA levels of CXCR4 in MM cell lines. The data are from three independent experiments and are presented as means ± standard deviation. D The representative 3D flow cytometry-based histograms depict CXCR4 expression on plasma cells (PC) in healthy donors (HD; black), monoclonal gammopathy of unknown significance (MGUS; light blue), smoldering MM (SMM; grey), newly diagnosed MM (NDMM; brown), and relapsed/refractory MM (RRMM; blue), compared to negative Ig isotype control (NEG; pink). The column bar graph shows the percentage of CXCR4 expression on normal PC (expressing CD138+/CD45+/CD38+) and non-PC of healthy donors (HD; n = 10), and malignant PC (expressing CD138+/CD45low/CD38++) and non-PC from primary bone marrow-derived cells of premalignant MGUS ( n = 15) and SMM ( n = 23), as well as active NDMM ( n = 39) and RMM ( n = 102) MM patients analyzed by flow cytometry and calculated using the Mann-Whitney U test, * p < 0.05 and **** p < 0.0001

Article Snippet: MM cell lines MM.1S (RRID:CVCL_8792) and U266 (RRID:CVCL_0566) were sourced from the American Type Culture Collection (ATCC, Manassas, VA), while OPM-2 (RRID:CVCL_1625) and L-363 (RRID:CVCL_1357) cell lines were acquired from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Braunschweig, Germany).

Techniques: Expressing, Flow Cytometry, Control, Western Blot, Reverse Transcription Polymerase Chain Reaction, Standard Deviation, Clinical Proteomics, Derivative Assay, MANN-WHITNEY

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Anti-myeloma activity of the CXCR4 antagonist WZ811

doi: 10.1007/s00109-026-02650-4

Figure Lengend Snippet: WZ811 decreases survival of MM cells. A MM cell lines (MM.1S, OPM-1, OPM-2, RPMI-S, RPMI-DOX6, RPMI-DOX40, RPMI-LR5, RPMI-MR20, JJN-3, KMS-11, L-363, OCI-My5, OCI-My7, and U266 cells) were exposed to indicated concentrations (0.625, 1.25, 2.5, 5, 10, 20, and 40 μM) of WZ811 for 48 h and 72 h. The cell survival was determined by MTT assay, and the EC 50 values of WZ811 were examined in MM cell lines for 48 h and 72 h using the CalcuSyn software. B Freshly sorted malignant bone marrow PC (expressing CD138+/CD45low/CD38++; n = 14) and tumor microenvironment cells (non-PCs; n = 14), both obtained from the same primary MM patient samples, were exposed to the indicated concentrations (1.25, 2.5, 5, 10, 20, and 40 μM) of WZ811 for 48 h. The cell survival was assessed by CellTiterGlo assay, and the EC 50 values of WZ811 in both PC and non-PC populations were determined for 48 h using the CalcuSyn software. If an exceptionally high EC 50 value was observed, it was reported as “n.a.” C Freshly isolated peripheral blood mononuclear cells (MNCs) from healthy donors (HD; n = 6) were exposed to indicated concentrations (0.625, 1.25, 2.5, 5, 10, 20, and 40 μM) of WZ811 for 48 h, and cell survival was assessed by MTT assay. The EC 50 values of WZ811 were determined in HD for 48 h using the CalcuSyn software. EC50 data are not presented due to exceptionally high values. Each treatment with a specific concentration of WZ811 was performed in triplicate. The presented data are mean ± standard deviation, expressed as survival/viability relative to untreated controls. EC 50 (half maximal effective concentration) is shown with lower and upper 95% confidence intervals (CI). D The median EC 50 values of WZ811 in MM cell lines, malignant PC, and non-PC of primary cells derived from MM patients, as well as MNCs of HD, were compared and calculated using One-way ANOVA on ranks followed by Kruskal-Wallis multiple comparison test, with *** p < 0.001

Article Snippet: MM cell lines MM.1S (RRID:CVCL_8792) and U266 (RRID:CVCL_0566) were sourced from the American Type Culture Collection (ATCC, Manassas, VA), while OPM-2 (RRID:CVCL_1625) and L-363 (RRID:CVCL_1357) cell lines were acquired from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Braunschweig, Germany).

Techniques: MTT Assay, Software, Expressing, Isolation, Concentration Assay, Standard Deviation, Derivative Assay, Comparison

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Anti-myeloma activity of the CXCR4 antagonist WZ811

doi: 10.1007/s00109-026-02650-4

Figure Lengend Snippet: WZ811 induces apoptosis and autophagy in MM cells. MM.1S, RPMI-S, and OPM-1 cells were treated with indicated concentrations (10, 20, 40, and 80 μM) of WZ811 for 72 h. A Representative flow cytometry-based dot plots showing the proportions of JC-1 aggregates versus JC-1 monomers in WZ811-treated MM.1S cells at 80 μM compared to control cells for 72 h. Decrease of mitochondrial membrane potential in MM cells after exposure to WZ811 was examined by staining with the fluorescent JC1 dye, indicating increased levels of JC-1 monomers, and analyzed by a FACS Canto II flow cytometer. B Representative flow cytometry-based dot plots showing the proportions of early apoptotic (Annexin V+/PI−), late apoptotic (Annexin V+/PI+/−), and necrotic (Annexin V+/PI+) cells in WZ811-treated MM.1S cells at 80 μM compared to control cells for 72 h. Induction of Annexin V+/Pi−, Annexin V+/Pi+/−, and Annexin V+/Pi+ cells was determined with Annexin V-FITC and Pi staining and analyzed by a FACS Canto II flow cytometer. The data are from three independent experiments and presented as means ± standard deviation. Significant differences between treatments and control were identified by one-way ANOVA followed by Dunnett’s multiple comparison test with * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. Modulation of C apoptotic- and D autophagic-related regulation proteins was evaluated in MM cells after exposure to WZ811 and compared to control (DMSO-treated) cells by Western blot analysis. Western blot analyses were performed on whole cell lysates (20 μg of proteins/lane) using anti-caspase-3, -caspase-7, -caspase-8, -PARP, -Bax, -AIF, -Mcl-1, -Beclin-1, -LC3A/B, -SQSTM1/p62, -ATG3, -ATG7, -ATG5-12, -ATG16L1, and -GAPDH (serving as a loading control) antibodies. Data are representative of two independent experiments

Article Snippet: MM cell lines MM.1S (RRID:CVCL_8792) and U266 (RRID:CVCL_0566) were sourced from the American Type Culture Collection (ATCC, Manassas, VA), while OPM-2 (RRID:CVCL_1625) and L-363 (RRID:CVCL_1357) cell lines were acquired from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Braunschweig, Germany).

Techniques: Flow Cytometry, Control, Membrane, Staining, Standard Deviation, Comparison, Western Blot

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Anti-myeloma activity of the CXCR4 antagonist WZ811

doi: 10.1007/s00109-026-02650-4

Figure Lengend Snippet: WZ811 induces G0/G1 cell cycle arrest in MM cells. A Flow cytometry-based cell cycle analysis of control and WZ811-treated MM.1S cells at 80 μM for 72 h with the distribution of cells in G0/G1, S, and G2/M phase showing the representative data out of three experiments. MM.1S, RPMI-S, and OPM-1 cells were exposed to indicated concentrations (10, 20, 40, and 80 μM) of WZ811 for 72 h, and their cell cycle profiles were analyzed using propidium iodide (Pi) staining. The distribution of cells in G0/G1, S, and G2/M phase was measured by a FACS Canto II flow cytometer and analyzed with De Novo FCS Express software. The data are from three independent experiments and are presented as means ± standard deviation. Significant differences between treatments and control were identified by one-way ANOVA followed by Dunnett’s multiple comparison test with * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001. B MM cell lines (MM.1S, RPMI-S, and OPM-1 cells) treated with WZ811 (10, 20, 40, and 80 μM) for 72 h were examined for changes in the levels of cell cycle regulatory and signaling molecules by western blot analysis. Western blot analyses of whole cell lysates (20 μg of proteins per lane) were immunoblotted using anti-mTOR, -p-mTOR, -ATM, -SIRT1, -c-Myc, -Notch1, -p-Cyclin B1, -Chk2, -Cdc2, -p-Cdc2, -histone H2AX (H2AX), -p-histone H2AX (p-H2AX), and -GAPDH (used as a loading control) antibodies. Results are representative of two independent experiments. C CXCR4 expression at the transcriptional mRNA level was examined after treatment with WZ811 (10, 20, 40, and 80 μM) for 6 h and 72 h in MM cell lines (MM.1S, RPMI-S, and OPM-1 cells) using RT-PCR analysis. Data are from three independent experiments and are presented as relative CXCR4 expression normalized to control ± standard deviation. D MM cell lines (MM.1S, RPMI-S, and OPM-1 cells) treated with WZ811 (10, 20, 40, and 80 μM) for 6 h were examined for changes in the expression levels of CXCR4 by western blot analysis. Western blot analyses of whole cell lysates (20 μg of proteins per lane) were immunoblotted using anti-CXCR4 and -GAPDH (used as a loading control) antibodies. Results are representative of two independent experiments

Article Snippet: MM cell lines MM.1S (RRID:CVCL_8792) and U266 (RRID:CVCL_0566) were sourced from the American Type Culture Collection (ATCC, Manassas, VA), while OPM-2 (RRID:CVCL_1625) and L-363 (RRID:CVCL_1357) cell lines were acquired from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Braunschweig, Germany).

Techniques: Flow Cytometry, Cell Cycle Assay, Control, Staining, Software, Standard Deviation, Comparison, Western Blot, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Anti-myeloma activity of the CXCR4 antagonist WZ811

doi: 10.1007/s00109-026-02650-4

Figure Lengend Snippet: WZ811 decreases the side population fractions and increases the expression extracellular matrix proteins CXCL12, collagen IV and laminin in MM cells. A RPMI-S cells were stained with carboxyfluorescein diacetate succinimidyl ester (CFSE) and cultured either alone or in the presence of BMSC HS-5 cells. Non-viable MM (CFSE+/7-AAD+) and HS-5 stromal (CFSE-/7-AAD+) cells were identified by 7-aminoactinomycin D (7-AAD) staining (upper row). Side population (SP) cells were determined by low intracellular staining with Hoechst 33342 fluorescence dye, gating only on CFSE+/7-AAD− RPMI-S cells, either alone or in the presence of BMSC HS-5 cells (middle row). The SP cell fraction was examined after treatment with 80 μM WZ811 for 24 h in RPMI-S cells, either alone or in the presence of BMSC HS-5 cells (lower row), using a FACS Aria Special Sorter UV laser flow cytometer (Becton Dickinson Biosciences, San Jose, CA, USA). B OPM-1 and RPMI-S cells, stained with CFSE, either alone or in the presence of BMSC HS-5 cells, were exposed to the indicated concentrations (10, 20, 40, and 80 μM) of WZ811 for 24 and 72 h. The normalized proportion of the SP cell fraction was calculated relative to the control (untreated) MM cells, both alone and in co-culture with BMSC HS-5 cells. C The expression of CXCL12, collagen IV, and laminin at the protein level in MM.1S, RPMI-S, and OPM-1 cells, stained with CFSE and treated with different concentrations of WZ811 (5, 10, 20, 40, and 80 μM) for 72 h, either alone or in the presence of unstained BMSC HS-5 cells, was analyzed by flow cytometry. The data are from three independent experiments performed in triplicate and are presented as means ± standard deviation. Significant differences between treatments and control were identified by one-way ANOVA followed by Dunnett's multiple comparison test, with * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001

Article Snippet: MM cell lines MM.1S (RRID:CVCL_8792) and U266 (RRID:CVCL_0566) were sourced from the American Type Culture Collection (ATCC, Manassas, VA), while OPM-2 (RRID:CVCL_1625) and L-363 (RRID:CVCL_1357) cell lines were acquired from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Braunschweig, Germany).

Techniques: Expressing, Staining, Cell Culture, Fluorescence, Flow Cytometry, Control, Co-Culture Assay, Standard Deviation, Comparison

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Anti-myeloma activity of the CXCR4 antagonist WZ811

doi: 10.1007/s00109-026-02650-4

Figure Lengend Snippet: WZ811 decreases tumor burden in a mouse xenograft model of human MM in vivo. A MM.1S cells (2.5 × 10 6 cells in 200 μl PBS) were injected subcutaneously into CB17/SCID mice, and treatment was initiated when tumors became palpable. Tumor-bearing mice were randomly assigned into two groups (six mice per group) treated with WZ811 (diluted in water for injection with 10% DMSO and 5% Tween 80) at a concentration 40 mg/kg or vehicle control (water with 10% DMSO and 5% Tween 80), both administered by oral gavage (once a day for 5 consecutive days followed by 2 days off, and repeated for 5 weeks) in the xenograft MM mouse model. Tumor diameters were measured every 2–3 days with a caliper, and tumor volumes were calculated according to the formula of the volume of an ellipse: V = 4/3π × ( a /2) × ( b /2) 2 , where a and b correspond to the longest and shortest tumor diameter, respectively. Representative photograph shows vehicle-treated tumors (upper row) versus WZ811-treated tumors (lower row) at the end of experiment using Pro 12MP camera system. B The mice treated with WZ811 showed significant suppression of tumor growth compared to the mice treated with vehicle. Significant differences in the volume of the tumors between WZ811- and vehicle-treated mice were evaluated by two-way ANOVA followed by Tukey multiple comparison test with * p = 0.0108. The data are presented as the means ± standard deviation. C Body weights of mice were measured every 2–3 days and compared between WZ811-treated and vehicle-treated mice. D Kaplan-Meier analysis of overall survival showed a significant increase in the survival of WZ811-treated mice compared to vehicle-treated control mice, calculated by Kaplan-Meier log-rank test, with *** p = 0.0008. Numbers at risk (# at risk), hazard ratio (HR), and 95% confidence interval (CI) are shown

Article Snippet: MM cell lines MM.1S (RRID:CVCL_8792) and U266 (RRID:CVCL_0566) were sourced from the American Type Culture Collection (ATCC, Manassas, VA), while OPM-2 (RRID:CVCL_1625) and L-363 (RRID:CVCL_1357) cell lines were acquired from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Braunschweig, Germany).

Techniques: In Vivo, Injection, Concentration Assay, Control, Comparison, Standard Deviation

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: Anti-myeloma activity of the CXCR4 antagonist WZ811

doi: 10.1007/s00109-026-02650-4

Figure Lengend Snippet: WZ811 in combination with anti-MM agents increases anti-MM activity in vitro. MM.1S, RPMI-S, and OPM-1 cells were exposed to the indicated concentrations (5, 10, and 20 μM) of WZ811 in combination with conventional agents: doxorubicin (DOX), dexamethasone (DEX) and melphalan (MEL), as well as novel agents: bortezomib (velcade; BTZ), carfilzomib (CFZ), lenalidomide (LEN), and pomalidomide (POM) anti-MM drugs for 24 h, 48 h and 72 h. Cell survival was then assessed by MTT assay, and combination effects were determined by the CalcuSyn software. A Fractions-affected (Fa; the ratio of the number of nonviable MM cells to the total number of MM cells) were visualized in a color-coded format and compared to treatment with each drug alone. B Isobologram analysis was performed to calculate the combination index (CI) for each combination by the Chou–Talalay method. The x-axis corresponds to the fractional effect at various combination doses in MM cells, and the y-axis represents the CI; CI < 1 indicates drug synergy, CI = 1 is considered additive, and CI > 1 indicates antagonism. All experiments were performed in triplicate. Data represent the median with interquartile range (IQR) of triplicate cultures

Article Snippet: MM cell lines MM.1S (RRID:CVCL_8792) and U266 (RRID:CVCL_0566) were sourced from the American Type Culture Collection (ATCC, Manassas, VA), while OPM-2 (RRID:CVCL_1625) and L-363 (RRID:CVCL_1357) cell lines were acquired from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSMZ, Braunschweig, Germany).

Techniques: Activity Assay, In Vitro, MTT Assay, Software

Journal: Blood Research

Article Title: Hypermethylation-mediated silencing of RASD1 drives multiple myeloma pathogenesis

doi: 10.1007/s44313-026-00125-6

Figure Lengend Snippet: a MSP gel electrophoresis showing methylation (M) and unmethylation (U) status in representative samples. b RASD1 mRNA expression in U266 cells following DAC treatment. c Quantification of apoptosis rates (mean ± SD) in U266 cells (** P < 0.01). d Representative flow cytometry plots of apoptosis (Annexin V-YF647A/PI staining); Total apoptosis: 5.04% in the control group and 12.08% in the DAC-treated group

Article Snippet: The human MM cell line U266 was obtained from the American Type Culture Collection, and was maintained in Dulbecco’s Modified Eagle Medium supplemented with 10% fetal bovine serum.

Techniques: Nucleic Acid Electrophoresis, Methylation, Expressing, Flow Cytometry, Staining, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: YMN-V115: a novel humanized BCMA/GPRC5D/CD3 trispecific antibody in relapsed/refractory multiple myeloma

doi: 10.1136/jitc-2025-013986

Figure Lengend Snippet: The expression of GPRC5D on r/r MM and generation of anti-GPRC5D clone 48# mAb. ( A ) The transcript expression levels of GPRC5D and BCMA in patients with MM (n=600) from MMRF-CoMMpass dataset. ( B–C ). Correlation analysis showing a low association between GPRC5D and BCMA expression (r=0.36). ( D ) Serum titers against GPRC5D in immunized mice (A90 and A91) measured 1 week after the final boost; recombinant B7H3-His served as a negative control. ( E ) Indirect ELISA demonstrating the binding activity of anti-GPRC5D antibodies to recombinant human GPRC5D protein. ( F ) Immunofluorescence microscopy (100×) confirming strong binding of anti-GPRC5D mAbs to GPRC5D-GFP-HeLa cells. Scale bars, 5 µm. ( G ) Flow cytometric analysis of antibody binding to MM cell lines (RPMI 8226, MM.1S) and negative control HEK293 cells. ( H ) Real-time cell analysis showing potent, GPRC5D-specific cytotoxicity of anti-GPRC5D mAbs. BCMA, B-cell maturation antigen; GPRC5D, G protein-coupled receptor class C group 5 member D; mAb, monoclonal antibody; MM, multiple myeloma; MMRF, Multiple Myeloma Research Foundation; r/r MM, relapsed/refractory multiple myeloma.

Article Snippet: The cell lines RPMI 8226, MM.1S, MM.1R, U937, and MV-4–11 were procured from the American Type Culture Collection (Manassas, USA).

Techniques: Expressing, Recombinant, Negative Control, Indirect ELISA, Binding Assay, Activity Assay, Immunofluorescence, Microscopy, Cell Analysis

Journal: Journal for Immunotherapy of Cancer

Article Title: YMN-V115: a novel humanized BCMA/GPRC5D/CD3 trispecific antibody in relapsed/refractory multiple myeloma

doi: 10.1136/jitc-2025-013986

Figure Lengend Snippet: Humanization and characterization of the anti-GPRC5D (clone 48#) mAb. ( A ) SDS-PAGE analysis confirmed the high purity of the humanized anti-GPRC5D scFv-hFc fusion proteins hu481-hFc and hu482-hFc. ( B ) Flow cytometric assessment of antibody binding to MM cell lines (RPMI 8226, MM.1S) and their GPRC5D knockout variants. ( C ) Immunofluorescence microscopy (100×) showing specific binding of hu482 to endogenous GPRC5D in MM.1S cells. Scale bar, 5 µm. ( D ) Indirect ELISA demonstrating comparable binding affinity of hu482 and the parental mu48 antibody. ( E ) Cross-species reactivity of hu482 analyzed by flow cytometry using HEK293T cells expressing human, cynomolgus, or murine GPRC5D. ( F ) Structural docking model of hu482 with GPRC5D predicted by ClusPro and visualized in PyMOL. ( G ) Schematic representation of GPRC5D chimeras highlighting the NTD, ECLs, TM, and CTD. ( H ) Flow cytometric evaluation of hu482 binding to HEK293T cells transfected with wild-type mGPRC5D or hGPRC5D mutant constructs. Representative results from two independent experiments are shown. CTD, C-terminal domain; DAPI, (4',6-Diamidino-2-phenylindole) dihydrochloride; ECLs, extracellular loops; GPRC5D, G protein-coupled receptor class C group 5 member D; mAb, monoclonal antibody; MM, multiple myeloma; NTD, N-terminal domain; scFv, single-chain variable fragment; SDS-PAGE, Sodium Dodecyl Sulfate PolyAcrylamide Gel Electrophoresis; TM, transmembrane.

Article Snippet: The cell lines RPMI 8226, MM.1S, MM.1R, U937, and MV-4–11 were procured from the American Type Culture Collection (Manassas, USA).

Techniques: SDS Page, Binding Assay, Knock-Out, Immunofluorescence, Microscopy, Indirect ELISA, Flow Cytometry, Expressing, Transfection, Mutagenesis, Construct, Polyacrylamide Gel Electrophoresis

Journal: Journal for Immunotherapy of Cancer

Article Title: YMN-V115: a novel humanized BCMA/GPRC5D/CD3 trispecific antibody in relapsed/refractory multiple myeloma

doi: 10.1136/jitc-2025-013986

Figure Lengend Snippet: Structural design and characterization of BCMA/GPRC5D/CD3 TriTEs. ( A ) Schematic representation of YMN-V115 and YMN-V118, trispecific antibodies targeting BCMA (green, clone:hu388), GPRC5D (blue, clone:hu482), and CD3 (orange, clone: OKT3) based on a knob-into-hole platform. ( B ) Size-exclusion chromatography analysis showing high purity with minimal aggregation. ( C–D ) Binding activity of YMN-V115 and YMN-V118 compared with isotype control in MM.1R ( C ) and RPMI 8226 ( D ) cell panels, including wild-type, BCMA-KO, GPRC5D-KO, and double-KO variants, confirming target-specific engagement of each arm. ( E–G ) Thermal stability of YMN-V115 assessed by Tm, Tagg, and DLS using the UNcle system (Unchained Labs). Data represent two independent experiments. ( H ) YMN-V115 maintained stable binding to MM.1R cells across serial dilutions of human serum. ( I ) sBCMA did not interfere with YMN-V115 binding to MM.1R cells. ( J ) Binding affinity remained unchanged in the presence of patient serum samples (n=3). APC, Allophycocyanin; BCMA, B-cell maturation antigen; DLS, dynamic light scattering; GPRC5D, G protein-coupled receptor class C group 5 member D; MFI, mean fluorescence intensity; MM, multiple myeloma; mAU, milli–absorbance units; sBCMA, soluble BCMA; Tagg, aggregation onset temperature; Tm, melting temperature; TriTE, trispecific T-cell engager.

Article Snippet: The cell lines RPMI 8226, MM.1S, MM.1R, U937, and MV-4–11 were procured from the American Type Culture Collection (Manassas, USA).

Techniques: Size-exclusion Chromatography, Binding Assay, Activity Assay, Control, Fluorescence

Journal: Journal for Immunotherapy of Cancer

Article Title: YMN-V115: a novel humanized BCMA/GPRC5D/CD3 trispecific antibody in relapsed/refractory multiple myeloma

doi: 10.1136/jitc-2025-013986

Figure Lengend Snippet: YMN-V115 shows superior antitumor activity to TCE benchmarks in the RPMI 8226 xenograft model of MM. ( A ) Schematic of the experimental procedure. Beginning on day 9, mice received different treatment regimens: PBS control, single bsAb treatment (JNJ-7957 or JNJ-7564), combination bsAb treatment (JNJ-7957/JNJ-7564), sequential treatment (bsAb first, followed by YMN-V115 on tumor relapse), or YMN-V115 alone (trispecific antibody, TriTE). All combination treatments were administered at a 1:1 ratio. ( B ) Bioluminescence imaging of the mice was conducted, with color gradients indicating luminescence intensity (red representing the highest and blue the lowest). ( C ) The quantification of average radiance (p/s/cm²/sr) of the luminescence is depicted. ( D ) Survival curves of RPMI 8226 xenograft mice treated with YMN-V115 and control formulations. ( E–G ). Serum proinflammatory cytokines in mice. After 1 day of treatment, the secretion levels of TNF-α, IFN-γ, and IL-2 in the MM.1S model. Representation of average fluorescence intensity in the limb ( H ) and in major organs, including the heart, liver, spleen, lung, kidney, and brain ( I ) on day 28. Data are presented as mean±SD. Statistical significance was assessed using one-way or two-way ANOVA, with significance levels indicated as follows: *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. ANOVA, analysis of variance; bsAb, bispecific antibody; IFN-γ, interferon-gamma; IL, interleukin; i.v., intravenous; MM, multiple myeloma; ns, not significant; PBS, phosphate-buffered saline; TCE, T-cell engager; TNF-α, tumor necrosis factor-alpha.

Article Snippet: The cell lines RPMI 8226, MM.1S, MM.1R, U937, and MV-4–11 were procured from the American Type Culture Collection (Manassas, USA).

Techniques: Activity Assay, Control, Imaging, Fluorescence, Saline