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: Advanced Science

Article Title: Coixenolide Enhances Antitumor Immunity of Cytotoxic CD8 + T Cells by Inhibiting S100A8/A9‐CD36 Axis in Obesity‐Associated Pancreatic Adenocarcinoma

doi: 10.1002/advs.202515759

Figure Lengend Snippet: Transcriptomic analyses revealed that coixenolide significantly inhibited S100a8/a9 + PAAD cells in HFD‐fed mice. (A) Subclusters of epithelial cells. (B) Malignancy score of epithelial cells. (C) The heterogeneity of epithelial cell clusters among different groups and tissue samples based on cell ratio. (D,E) Changes in the proportions of Cluster 2 (D) and Cluster 8 (E) tumor cells. n = 7 for HFD group and n = 6 for CD group. Data are presented as mean ± SD; a two‐sided t ‐test was used. (F) The upregulated genes across 21 tumor cell subsets. (G) Pathway enrichment associated with the top 100 upregulated genes (ranked by p ‐value) of Clusters 2 and 8 tumor cells. (H) CytoTRACE score and distribution of tumor cell clusters. (I) Shared genes of Clusters 2 and 8. (J) The colocalization of S100A8/A9 (yellow), Pan‐CK (red), and DAPI (deep blue) in the tumor tissues of CD‐ and HFD‐fed mice; representative of three independent experiments. (K) The expression patterns of S100A8 and S100A9 between tumor and normal pancreatic tissues using the GEPIA webtool; n = 179 (tumor tissue) and 171 (normal tissue); a two‐sided Wilcoxon test was used. (L) The colocalization of S100A8/A9 (yellow), insulin (pink), Pan‐CK (white), and DAPI (deep blue) in the peritumoral pancreatic tissue, as well as in tumor tissues in PAAD patients. (M,N) Kaplan–Meier survival plot indicating that patients with high levels of S100A8 (M) or S100A9 (N) have a worse clinical outcome in the TCGA PAAD cohort; log‐rank test was used. (O,P) The expressions of S100A8 (O) and S100A9 (P) between patients with PAAD of different invasion levels, grades, and AJCC stages. The data were sourced from the TCGA cohort; a two‐sided t ‐test was used. (Q) Expression levels of S100a8 and S100a9 in Clusters 2 and 8 tumors of HFD‐fed mice following coixenolide treatment. A two‐sided t ‐test was used. (R,S) Western blot (R) and flow cytometry plot (S) measuring S100a8/a9 expression in Pan02 cells following coixenolide treatment. n = 4 independent experiments; two‐sided t ‐test was used. (T) The levels of S100a8/a9 in the supernatants of Pan02 cells with or without coixenolide treatment were determined using ELISA. n = 3 independent experiments; two‐sided t ‐test was used. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The CD8 + T cell supernatants were collected, and the secretion of TNF‐α (Mouse TNF‐alpha ELISA Kit, RK00027, Abclonal), IFN‐γ (Mouse IFN‐gamma ELISA Kit, RK00019, Abclonal), and GZMB (Mouse Granzymes B ELISA Kit, RK00007, Abclonal) was quantified using specific ELISA kits following the standard protocols.

Techniques: Expressing, Western Blot, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: Coixenolide Enhances Antitumor Immunity of Cytotoxic CD8 + T Cells by Inhibiting S100A8/A9‐CD36 Axis in Obesity‐Associated Pancreatic Adenocarcinoma

doi: 10.1002/advs.202515759

Figure Lengend Snippet: Cell–cell communication analysis indicated that S100a8/a9 served as a critical target for coixenolide, regulating the interactions between PAAD cells and CD8 + T cells. (A) The overview of cell–cell interactions between malignant epithelial cells and T cells in the tumor tissues treated with vehicle control and coixenolide, respectively. (B) The intensity of intercellular signaling, showing both the sent and received signals. The cells in the upper right corner exhibit the strongest incoming and outgoing interactions. S100a8/a9 + tumor cell clusters (Clusters 2 and 8) are highlighted in red boxes. Cytotoxic CD8 + T and Treg cells are indicated by red arrows. (C) The outgoing communication patterns of epithelial cells and T cells in the tumor of HFD‐fed mice with or without coixenolide treatment. (D) CD8 + T cell killing assays were conducted using coculture of Pan02 and CD8 + T cells. Pan02 or CD8 + T cells were pretreated with 25 mg/mL coixenolide before coculture. (E) Primary CD8 + T cells were activated by incubation with CD3/CD28 antibody and labeled with carboxy‐fluorescein succinimidyl ester (CFSE). (F–H) The levels of TNF‐α (F), IFN‐γ (G), and Granzyme B (GZMB) (H) in the supernatants after co‐culture of CD8 + T and Pan02 cells pretreated with coixenolide (+) or vehicle control (−) were determined using ELISA. (I–K) The expression of TNF‐α (I), IFN‐γ (J), and Granzyme B (GZMB) (K) in CD8 + T cells after co‐culture of CD8 + T cells and Pan02 cells pretreated with coixenolide (+) or vehicle control (−) was determined using flow cytometry. (L) Flow cytometry analyses of T cell exhaustion markers (CTLA‐4) on CD8 + T cells after coculture of Pan02 and CD8 + T cells pretreated with coixenolide (+) or vehicle control (−). (M,N) Flow cytometry analyses of CD8 + T cell killing (M) and division (N) activities. CD8 + T cells were cocultured with S100a9 knockout (KO) or wild‐type (WT) Pan02 cells. (O–Q) The levels of TNF‐α (O), IFN‐γ (P), and GZMB (Q) in the supernatants after coculture of CD8 + T cells with WT or S100a9 KO Pan02 cells were determined using ELISA. n = 3 or 4 independent coculture experiments or replicates. One‐way ANOVA with Tukey's post‐hoc test (E–L), two‐sided t ‐test (M–N), or two‐way ANOVA (O‐Q) was used. All p values are two‐sided. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The CD8 + T cell supernatants were collected, and the secretion of TNF‐α (Mouse TNF‐alpha ELISA Kit, RK00027, Abclonal), IFN‐γ (Mouse IFN‐gamma ELISA Kit, RK00019, Abclonal), and GZMB (Mouse Granzymes B ELISA Kit, RK00007, Abclonal) was quantified using specific ELISA kits following the standard protocols.

Techniques: Control, Incubation, Labeling, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry, Knock-Out

Journal: Cell Reports Medicine

Article Title: A biomimetic hybrid membrane vesicle nanoplatform attenuates tendinopathy through neuroinflammation modulation and tendon regeneration

doi: 10.1016/j.xcrm.2026.102937

Figure Lengend Snippet: MSM-DTM adsorbs inflammatory cytokines, suppresses TSPCs inflammation and senescence, and promotes tenogenic differentiation and migration (A) The remaining concentrations of TNF-α, IL-1β, IL-6, IFN-γ, LPS, and NGF were detected by ELISA after co-culture with different concentrations of MSM. (B and C) RT-qPCR of IL-6 and CCL-2 in TSPCs under different treatments. (D) Western blot of COL1A2, MMP3, and TNMD. (E) ELISA of TNF-α, IL-6, and IFN-γ in supernatants. (F)Western blot of p16 and p21 after 7 days of treatment. (G) β-Galactosidase staining analysis after 7 days of treatment. Scale bars, 100 μm. (H and I) Immunofluorescence and quantification of TNMD and TNC after 14-day treatment. Scale bars, 50 μm. (J) Relative mRNA expression of tenogenic differentiation markers SCX and TNC. (K and L) Representative images and quantification of Transwell assay. Scale bars, 100 μm. (M and N) Representative images and quantification of wound healing assay. Scale bars, 100 μm. Statistical comparisons were performed with one-way ANOVA with Tukey’s multiple comparisons test. Data are presented as the mean ± SD, n = 3. Ns, no significance, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 between groups.

Article Snippet: IFN-γ ELISA kit , ABclonal , RK00015.

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Quantitative RT-PCR, Western Blot, Staining, Immunofluorescence, Expressing, Transwell Assay, Wound Healing Assay

Journal: Cell Reports Medicine

Article Title: A biomimetic hybrid membrane vesicle nanoplatform attenuates tendinopathy through neuroinflammation modulation and tendon regeneration

doi: 10.1016/j.xcrm.2026.102937

Figure Lengend Snippet:

Article Snippet: IFN-γ ELISA kit , ABclonal , RK00015.

Techniques: Recombinant, Lysis, Hydroxyproline Assay, Enzyme-linked Immunosorbent Assay, Isolation, SYBR Green Assay, Bicinchoninic Acid Protein Assay, Staining, Software

Journal: Advanced Science

Article Title: Senkyunolide I Inhibits mtDNA‐cGAS‐STING Signaling in Macrophages via Targeting VDAC1 Oligomerization to Attenuate Ulcerative Colitis

doi: 10.1002/advs.77045

Figure Lengend Snippet: ABPP proteomic analysis confirms VDAC1 as target protein of SEI. (A) Chemical structures of SEI and SEI probe (SEI‐P). (B) Cell viability of iBMDM cells treated with SEI or SEI‐P. (C) Release of inflammatory cytokine IL‐1β and (D) cell viability in TNF‐α plus IFN‐γ‐induced iBMDM cells. (E) Cellular imaging of SEI‐P with different exposure times in iBMDM cells. (F) Dose‐dependent labeling of proteins by SEI‐P in iBMDM cells. (G) Competition between SEI and SEI‐P for protein binding in situ (red star: 35 kD). (H) Chemical proteomics analysis workflow for identifying potential targets of SEI, created using Figdraw. (I) Volcano plot of proteins identified in the ABPP method. The graph displayed the log 2 FC of the competition group (100 µ m SEI + 50 µ m SEI‐P) versus SEI‐P (50 µ m ) ( x ‐axis) against the −log 10 ( p‐ value) ( y ‐axis). Among these, points with p < 0.05 and log 2 FC < −1 (blue) were selected as target protein candidates. Values were expressed as mean ± SD ( n = 3).

Article Snippet: Senkyunolide I (HY‐N0745), Erastin (HY‐15763), H‐151 (HY‐112693), TNF‐α (HY‐P7058 or HY‐P7090), IFN‐γ (HY‐P7025 or HY‐P7071), Lipopolysaccharides (LPS, HY‐D1056), Adenosine 5'‐triphosphate (ATP, HY‐B2176) were purchased from MedChem Express.

Techniques: Imaging, Labeling, Protein Binding, In Situ

Journal: Advanced Science

Article Title: Senkyunolide I Inhibits mtDNA‐cGAS‐STING Signaling in Macrophages via Targeting VDAC1 Oligomerization to Attenuate Ulcerative Colitis

doi: 10.1002/advs.77045

Figure Lengend Snippet: SEI directly interacts with VDAC1 to inhibit its oligomerization. (A) The interaction between SEI and VDAC1 was detected using molecular docking. (B) Lysates from iBMDM cells were incubated with or without SEI (100 µ m ) for 24 h. Different concentrations of pronase E were added for 20 min, and VDAC1 content was analyzed using WB analysis. (C) Lysates from iBMDM cells were incubated with SEI at the indicated concentrations for 24 h, with a final concentration of 0.01% pronase E added for 20 min. The level of VDAC1 was assessed through WB analysis. (D) iBMDM cells were incubated with SEI (100 µ m ) for 24 h. These samples were then analyzed using CETSA. Values were expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01 versus SEI group. (E) MST demonstrating a direct interaction between SEI and EGFP‐tagged VDAC1 in lysates from EGFP‐VDAC1 expressing HEK293T cells. (F) Immunoblotting analysis of VDAC1 cross‐linking in iBMDM cells, untreated or stimulated with TNF‐α plus IFN‐γ, with or without addition of SEI (100 µ m ). (G) The VDAC1‐OE plasmid was transfected into iBMDM cells, and cGAMP production was subsequently measured by ELISA. (H) iBMDM cells were transfected with VDAC1‐OE, and then the phosphorylation levels of STING and IRF3 were measured by WB. (I) The mRNA expression levels of CCL5 , CXCL10 , and ISG15 in iBMDM cells transfected with VDAC1‐OE were measured by qRT‐PCR. (J) qRT‐PCR analysis of cytoplasmic mtDNA ( mt‐Nd1 , D‐loop and mt‐Cytb ). Values were expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Senkyunolide I (HY‐N0745), Erastin (HY‐15763), H‐151 (HY‐112693), TNF‐α (HY‐P7058 or HY‐P7090), IFN‐γ (HY‐P7025 or HY‐P7071), Lipopolysaccharides (LPS, HY‐D1056), Adenosine 5'‐triphosphate (ATP, HY‐B2176) were purchased from MedChem Express.

Techniques: Incubation, Concentration Assay, Expressing, Western Blot, Plasmid Preparation, Transfection, Enzyme-linked Immunosorbent Assay, Phospho-proteomics, Quantitative RT-PCR

Journal: Advanced Science

Article Title: Senkyunolide I Inhibits mtDNA‐cGAS‐STING Signaling in Macrophages via Targeting VDAC1 Oligomerization to Attenuate Ulcerative Colitis

doi: 10.1002/advs.77045

Figure Lengend Snippet: SEI directly binds to K12 on VDAC1. (A) The RMSD of the VDAC1 backbone was simulated for a range of 100 ns. (B) The RMSF values of all amino acid residues were simulated. (C) Radius of gyration of the apo (blue) and protein when bound to the ligand (red) for the 100 ns simulation. (D) DCCM analysis matrix of VDAC1 protein; the region in red indicates residue pairs in horizontal and vertical coordinates have positive correlation in movement patterns, while the region in blue indicates negative correlation. (E) DCCM analysis matrix of SEI‐VDAC1 complex. (F–H) Movement correlation of each residue with a negative correlation coefficient ranged from −0.6 to −0.8 of VDAC1 protein (F), ranged from −0.4 to −0.6 of VDAC1 protein (G), and ranged from −0.4 to −0.6 of SEI‐VDAC1 complex (H). (I) The total binding free energy was calculated, and a series of contribution components were analyzed. Data are presented as energy changes in units of kJ/mol. (J) Ten residues of the VDAC1‐SEI complex were selected and analyzed. Data were presented as energy changes in a unit of kJ/mol with different contributors indicated by colors. (K) Free energy landscape. (L) Sequence conservation analysis of VDAC1 protein using ESPript 3.0. (M) The iBMDM cells were transfected with K174, K12, and G172 mutation plasmids and then treated with DMSO or SEI (100 µ m ) for 1 h. The interaction between SEI and VDAC1 was detected using the CETSA assay. Values were expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 versus Ctrl group. (N) VDAC1 −/− iBMDM cells were transfected with Flag‐VDAC1(WT), Flag‐VDAC1(K174A), Flag‐VDAC1(K12A), or Flag‐VDAC1(G172A). Immunoblotting analysis of VDAC1 cross‐linking in iBMDM cells stimulated with TNF‐α plus IFN‐γ, with or without addition of SEI (100 µ m ). (O) VDAC1 −/− iBMDM cells were transfected with an empty vector, Flag‐VDAC1‐WT, Flag‐VDAC1(K174A), Flag‐VDAC1(K12A), or Flag‐VDAC1(G172A). WB analyses of the quantity of p‐TBK1 and p‐IRF3 after stimulation with TNF‐α plus IFN‐γ and treatment with SEI (100 µ m ) or left untreated (control) for 24 h. Values were expressed as mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Senkyunolide I (HY‐N0745), Erastin (HY‐15763), H‐151 (HY‐112693), TNF‐α (HY‐P7058 or HY‐P7090), IFN‐γ (HY‐P7025 or HY‐P7071), Lipopolysaccharides (LPS, HY‐D1056), Adenosine 5'‐triphosphate (ATP, HY‐B2176) were purchased from MedChem Express.

Techniques: Residue, Binding Assay, Sequencing, Transfection, Mutagenesis, Western Blot, Plasmid Preparation, Control