Journal: International Journal of Molecular Sciences

Article Title: Targeting ADAR1 Restores Interferon Signaling and Enhances Immunotherapy Response in Multiple Myeloma

doi: 10.3390/ijms27156602

Figure Lengend Snippet: Single-cell transcriptomic analysis showing ADAR1 expression patterns and its association with IFN signaling in multiple myeloma. ( A ) UMAP visualization of single-cell RNA-seq data from the GSE124310 dataset showing major cell populations in the MM bone marrow microenvironment. ( B ) Feature plot showing the distribution of ADAR1 expression across different cell clusters. ( C ) Violin plot comparing ADAR1 expression levels among different immune cell populations. ( D ) Comparison of ADAR1 expression between malignant plasma cells and plasma cells from healthy donors. ( E ) Analysis of T-cell exhaustion signatures in MM samples. ( F ) Type I IFN pathway activity in malignant plasma cells stratified by ADAR1 expression levels.

Article Snippet: For IFN stimulation, recombinant human IFNα (10 ng/mL; MedChemExpress, HY- P78672 , Monmouth Junction, NJ, USA) was added to the co-culture system and incubated for 48 h. For immune checkpoint inhibition, pembrolizumab (20 μg/mL; MedChemExpress, Cat. No. HY-P9902) was applied for 48 h. Control groups received PBS or isotype control antibodies.

Techniques: Single Cell, Expressing, RNA Sequencing, Comparison, Clinical Proteomics, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Targeting ADAR1 Restores Interferon Signaling and Enhances Immunotherapy Response in Multiple Myeloma

doi: 10.3390/ijms27156602

Figure Lengend Snippet: ADAR1 suppresses MDA5-mediated type I interferon signaling in multiple myeloma. ( A ) Comparison of A-to-G RNA editing frequency between ADAR1-high and ADAR1-low samples in the GSE110486 dataset. ( B ) RIP-qPCR analysis showing the enrichment of Alu dsRNA associated with ADAR1 and MDA5 following ADAR1 knockdown. ( C ) Western blot validation of MDA5 immunoprecipitation in the RIP assay. ( D ) Western blot analysis of IFNα and IFNβ expression after ADAR1 knockdown in U266 and H929 cells co-cultured with HS-5 stromal cells. ( E ) ELISA measurement of IFNα and IFNβ secretion following ADAR1 knockdown. ( F ) Comparison of IFNα levels in peripheral blood samples from healthy donors and MM patients. ( G ) Spearman correlation analysis between ADAR1 expression and IFNα levels in MM patient samples. ( H ) Western blot analysis of IFNα, IFNβ, and ISG15 expression following simultaneous knockdown of ADAR1 and MDA5. ( I ) ELISA measurement of IFNα and IFNβ secretion after co-silencing ADAR1 and MDA5. ( J ) Western blot analysis of ISG15 expression following combined knockdown of ADAR1 and STAT1. ( K ) ELISA analysis of IFNα and IFNβ secretion after STAT1 knockdown. ns, not significant; * p < 0.05; *** p < 0.001.

Article Snippet: For IFN stimulation, recombinant human IFNα (10 ng/mL; MedChemExpress, HY- P78672 , Monmouth Junction, NJ, USA) was added to the co-culture system and incubated for 48 h. For immune checkpoint inhibition, pembrolizumab (20 μg/mL; MedChemExpress, Cat. No. HY-P9902) was applied for 48 h. Control groups received PBS or isotype control antibodies.

Techniques: Comparison, Knockdown, Western Blot, Biomarker Discovery, Immunoprecipitation, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Targeting ADAR1 Restores Interferon Signaling and Enhances Immunotherapy Response in Multiple Myeloma

doi: 10.3390/ijms27156602

Figure Lengend Snippet: The ADAR1–MDA5–IFNα axis regulates CD8 + T-cell activity and affects the response of multiple myeloma cells to PD-1 blockade. ( A ) ELISA analysis of IFNα and IFNβ secretion in U266 monoculture and U266–HS-5 co-culture systems. ( B ) CCK-8 assay measuring the relative CD8 + T-cell proliferation in the triple co-culture system. T-cell proliferation in the NC group was normalized to 100%, and values in the remaining groups are presented relative to the NC control. ( C ) ELISA analysis of PF1 and GZMB secretion in the triple co-culture system. ( D ) Spearman correlation analysis between IFNα levels and CD8 + T-cell proliferation and cytotoxic molecule expression. ( E ) CCK-8 assay evaluating U266 cell proliferation under different experimental conditions. ( F ) Flow cytometric analysis of U266 cell apoptosis using Annexin V/PI staining. ( G ) Flow cytometric analysis of U266 cell apoptosis, including early, late, and total apoptosis. ns, not significant; * p < 0.05.

Article Snippet: For IFN stimulation, recombinant human IFNα (10 ng/mL; MedChemExpress, HY- P78672 , Monmouth Junction, NJ, USA) was added to the co-culture system and incubated for 48 h. For immune checkpoint inhibition, pembrolizumab (20 μg/mL; MedChemExpress, Cat. No. HY-P9902) was applied for 48 h. Control groups received PBS or isotype control antibodies.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, CCK-8 Assay, Control, Expressing, Staining

Journal: International Journal of Molecular Sciences

Article Title: Targeting ADAR1 Restores Interferon Signaling and Enhances Immunotherapy Response in Multiple Myeloma

doi: 10.3390/ijms27156602

Figure Lengend Snippet: ADAR1 inhibition enhances the antitumor efficacy of PD-1 blockade in a multiple myeloma mouse model. ( A ) Endpoint tumor volumes in the MOPC315/BALB/c mouse model across different treatment groups. ( B ) Tumor growth curves of mice treated with NC, 8-azaadenosine, PD-1 blockade, or combination therapy. ( C ) Representative TUNEL staining of tumor sections showing apoptotic cells (red) with DAPI nuclear counterstaining (blue). ( D ) Western blot analysis of IFNα protein expression in tumor tissues. ( E ) Immunohistochemical staining showing CD8 + T-cell infiltration in tumor tissues. ** p < 0.01.

Article Snippet: For IFN stimulation, recombinant human IFNα (10 ng/mL; MedChemExpress, HY- P78672 , Monmouth Junction, NJ, USA) was added to the co-culture system and incubated for 48 h. For immune checkpoint inhibition, pembrolizumab (20 μg/mL; MedChemExpress, Cat. No. HY-P9902) was applied for 48 h. Control groups received PBS or isotype control antibodies.

Techniques: Inhibition, TUNEL Assay, Staining, Western Blot, Expressing, Immunohistochemical staining

Journal: International Journal of Molecular Sciences

Article Title: Targeting ADAR1 Restores Interferon Signaling and Enhances Immunotherapy Response in Multiple Myeloma

doi: 10.3390/ijms27156602

Figure Lengend Snippet: Proposed model of ADAR1-mediated immune evasion in multiple myeloma. ADAR1 promotes A-to-I editing of endogenous dsRNA, thereby limiting its recognition by the innate immune sensor MDA5. Suppression of the MDA5–MAVS pathway attenuates type I interferon signaling, resulting in reduced IFN-α production and impaired downstream JAK/STAT activation. Consequently, CD8 + T-cell function is compromised, contributing to the establishment of an immunosuppressive bone marrow microenvironment.

Article Snippet: For IFN stimulation, recombinant human IFNα (10 ng/mL; MedChemExpress, HY- P78672 , Monmouth Junction, NJ, USA) was added to the co-culture system and incubated for 48 h. For immune checkpoint inhibition, pembrolizumab (20 μg/mL; MedChemExpress, Cat. No. HY-P9902) was applied for 48 h. Control groups received PBS or isotype control antibodies.

Techniques: Activation Assay, Cell Function Assay

Journal: bioRxiv

Article Title: Human antibodies against West Nile and related orthoflaviviruses

doi: 10.64898/2026.04.02.715800

Figure Lengend Snippet: (A) IgG binding to WNV EDIII by serial serum dilution. ELISA curves for samples from the 72 WND/WNF-suspected individuals and 3 orthoflavivirus naïve controls (black) are shown. The average of two independent experiments is shown. (B) Serum neutralization screening with WNV RVP . Shown is the rank-ordered NanoLuc activity relative to no-serum control for the 72 WND/WNF-suspected individuals and 1 orthoflavivirus naïve control (black); lower values correspond to higher neutralization. Samples with relative NanoLuc signal below 0.3 (dotted line) were selected for neutralization curves in (C). Each bar represents and individual participant sample analyzed at 1:100 dilution. Average signal of triplicate wells from a single experiment. (C) Neutralization of WNV RVP by serial serum dilution. Shown is the NanoLuc activity relative to no-serum control for 36 WND/WNF cases and 1 orthoflavivirus naïve control (black). Mean ± SD of triplicates. Representative of two independent experiments. (D-F) Identification of serum autoantibodies to IFN-α2 and IFN-ω. Plots compare the ability of serum IgG to bind, and of serum to neutralize, IFN-α2 (D) and IFN-ω (E) (n=39; only samples for which sufficient serum was available were assayed). The comparison of IFN-α2 and IFN-ω neutralization is shown in (F). Binding is shown as relative Mean Fluorescence Intensity (MFI) and neutralization as relative ISG15-promoter driven luciferase signal compared to no serum control. The dotted lines indicate the threshold for positivity of the assay. , Representative of 2 independent experiments. (G) Age and gender distribution of the study participants with or without IFN-α2 and/or IFN-ω neutralizing autoantibodies. Welch’s test. (H and I) Serum IgG binding to WNV EDIII and serum neutralization of WNV RVP in study participants with or without IFN-α2 and/or IFN-ω neutralizing autoantibodies. The two groups are compared with respect to (H) IgG binding to WNV EDIII (Area Under the Curve (AUC) of ELISA and (I) neutralization of WNV RVP (NT 50 values). Horizontal lines indicate the mean. Mann-Whitney test. In (A to C), green, blue and red indicate samples from WNV-infected individuals from which antibodies were derived (Figure S2B and C). In (D to I), female is circle, male is square, dark blue neutralizes both IFN-α2 and IFN-ω, light blue neutralizes IFN-α2 only, grey does not neutralize either.

Article Snippet: Serum samples were diluted 1:20 in OptiMEM (Optimized Minimal Essential Medium; Gibco, 31985070) containing 0.01 ng/mL IFN-α2 (Novus Biologicals, NBP2-34971) or 0.02 ng/mL IFN-ω (Novus Biologicals, NBP2-35893), together with a live-cell Renilla luciferase substrate (EnduRen, Promega, E6481; 1:10,000), and incubated for 1 hour with constant shaking at 600 rpm.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Neutralization, Activity Assay, Control, Comparison, Fluorescence, Luciferase, MANN-WHITNEY, Infection, Derivative Assay