Journal: Journal of Human Immunity

Article Title: Adult-onset STING-associated vasculopathy

doi: 10.70962/jhi.20250235

Figure Lengend Snippet: Clinical and structural basis of AO-SAVI caused by STING gain-of-function mutations. (A) Representative images of cutaneous vasculopathy in a patient with STING R281Q, showing purpuric rashes (left) and ulcerated skin lesions with biopsies revealing thrombotic vasculopathy (right). (B) Top: Schematic of the linear domain architecture of STING with locations of V155M (connector loop) and R281Q (polymeric interface) mutations. Bottom: Schematic of conformational states; wild-type (dimer unrotated, C-terminal blocked), V155M (dimer rotation, C-terminal release), and R281Q (dimer unrotated, C-terminal release). Created with BioRender.com , released under a CC-BY 4.0 license. (C) Human WT STING dimer in the apo state (PBD: 6NT5 ). On the left, the N-terminal transmembrane domain (TMD) (bottom, blue), connector loop (CL) (middle, green), and ligand-binding domain (LBD) (top, purple) are indicated. Insets show V155 and R281 position. (D) Crystal lattice of six human STING (H232) monomers bound to cGAMP (PBD: 4LOH ), denoting polymeric interface (black dotted line) and the interaction between residue R281 (red) and D301 (blue). Crystal packing patterns are thought to mimic native STING polymerization . (E) Chest computed tomography image demonstrating interstitial lung disease in a symptomatic patient with the STING V155M mutant (patient 4). (F) Brain MRI (T2-weighted image) showing demyelinating CNS inflammatory lesions in a symptomatic patient with the STING V155M mutation (patient 5). (G) Plasma IFN-α concentrations measured by digital ELISA in healthy control individuals (HC, n = 12), AGS ( n = 16), and individuals with STING1 variants identified through the PMBB ( n = 5). Each point represents a single patient, and the dotted line indicates three times the standard deviation above the HC mean. Blue triangles denote asymptomatic individuals; orange circles denote symptomatic patients on immunosuppressive therapy. cGAMP, cyclic GMP-AMP.

Article Snippet: The Simoa IFN-α MS advantage PLUS kit was purchased from Quanterix and run by the Human Immunology Core at the University of Pennsylvania on a Simoa HD-X machine according to the manufacturer’s recommendations.

Techniques: Ligand Binding Assay, Residue, Computed Tomography, Mutagenesis, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Control, Standard Deviation

Journal: Molecular Therapy Advances

Article Title: Efficacy and safety of SENS-501, a dual-AAV otoferlin gene therapy, for DFNB9 congenital deafness

doi: 10.1016/j.omta.2026.201762

Figure Lengend Snippet: Intracochlear injection of SENS-501 in NHPs results in a mild humoral and an undetectable cellular response to the capsid (A) AAV8 anti-drug antibody (ADA) titers measured in serum at pre-dose and at 16, 29, and 92 days post-vector administration in NHPs. (B) Anti-AAV8 neutralizing antibodies (NAb) titers measured in serum at pre-dose and at 16, 29, and 92 days post-vector administration in NHPs. (C and D) IFN-γ spot forming units (SFUs) measured by ELISpot assay at 29 (C; left) and 92 (D; right) days post-injection. Peripheral blood mononuclear cells (PBMCs) from the indicated groups were stimulated with three different AAV8 peptide pools and a positive control (PMA/ionomycin). The dotted line represents the assay-specific positivity threshold. Each dot represents one animal. Bars represent the mean ± SEM.

Article Snippet: After the incubation, detection was performed with a monoclonal anti-monkey IFN-γ antibody (Monkey IFN-γ ELISpot Pro Kit, Mabtech) coupled with alkaline phosphatase and incubated with BCIP/NBT (5-bromo-4-chloro-3-indolyl-1-phosphate / nitroblue tetrazolium) substrate to detect secreted IFN-γ.

Techniques: Injection, Plasmid Preparation, Enzyme-linked Immunospot, Positive Control

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: Smart Molecules

Article Title: Endocytosis‐independent cytosolic entry of messenger RNA via fluorous bilayer zippering attenuating Toll‐like receptor signaling and enables ischemic tissue salvage

doi: 10.1002/smo2.70085

Figure Lengend Snippet: Biocompatibilities and immunogenicity of mRNA‐encapsulating polyplexes. (a) Cell viabilities of HUVECs upon 24 h incubation in presence of a variety of mRNA‐encapsulating polyplexes (mean ± s.d., n = 4, ** p < 0.01, student t test). (b) Hemolytic activities of sheep red blood cells upon 2 h incubation with varied concentrated delivery materials (*** p < 0.005; Student's t ‐test, mean ± s.d., n = 4). (c) confocal laser scanning microscopy (CLSM) measurement for assessment of overall cellular internalization of mRNA‐encapsulating polyplexes into RAW264.7 cells. (d) Interferon‐β (IFN‐β): at 4 h post incubation, the expression levels of inflammatory molecules were measured with qRT‐PCR. (e) Interleukin‐8 (IL‐8): at 4 h post incubation, the expression levels of inflammatory molecules were measured with qRT‐PCR. (** p < 0.01, *** p < 0.005; Student's t ‐test, n = 4).

Article Snippet: Culture supernatants (100 μL/well) were harvested and assayed for IFN‐α using the Mouse IFN‐α ELISA Kit (PBL Interferon Source) according to the manufacturer's instructions.

Techniques: Immunopeptidomics, Incubation, Confocal Laser Scanning Microscopy, Expressing, Quantitative RT-PCR

Journal: Smart Molecules

Article Title: Endocytosis‐independent cytosolic entry of messenger RNA via fluorous bilayer zippering attenuating Toll‐like receptor signaling and enables ischemic tissue salvage

doi: 10.1002/smo2.70085

Figure Lengend Snippet: Revascularization in hindlimbs by local dosage of mVEGF‐encapsulating polyplexes. (a) Therapeutic scheme. (b) Anatomy of the established hindlimb ischemia model. Ligations were made in the femoral artery at the proximal and distal sites. (c) Angiogenesis in mouse hindlimbs post ligation. (d) Visualization of blood flow by Laser Speckle Flowgraphy on Day 28 post‐dosage of mVEGF therapeutics (mVEGF: 10 μg). The magnified inset images captured by intravital confocal laser scanning microscopy (CLSM), revealing vasculature details by intravenous dosage of FITC‐dextran (MW: 10 kDa). (e) Estimation of blood perfusion volume based on quantification by laser speckle flowgraphy on Day 28 post‐dosage of mVEGF therapeutics (mVEGF: 10 μg). The data were represented as the mean ± standard deviations (s.d.) ( n = 5). (* p < 0.05, ** p < 0.01, student t test). (f) Quantification of the expressed VEGF protein on day 4 post dosage by ELISA.

Article Snippet: Culture supernatants (100 μL/well) were harvested and assayed for IFN‐α using the Mouse IFN‐α ELISA Kit (PBL Interferon Source) according to the manufacturer's instructions.

Techniques: Ligation, Confocal Laser Scanning Microscopy, Enzyme-linked Immunosorbent Assay