Journal: MedComm

Article Title: An Engineered Extracellular Vesicle With Enhanced Tumor and Lymph Nodes Targeting as siRNA Delivery System for Robust Tumor Immunotherapy

doi: 10.1002/mco2.70673

Figure Lengend Snippet: DP‐EVs exhibit enhanced accumulation in tumors and lymph nodes. (A) Experimental timeline of in vivo biodistribution. (B and C) Fluorescence imaging of normal saline (NS group), DIR‐labeled EVs and DP‐EVs 24 h postinjection in E.G7‐OVA tumor bearing mice. (D) Representative CLSM images of frozen sections of major organs after subcutaneous injection of DIR‐EVs and DIR‐DP‐EVs. Scale bars, 100 µm. (E and F) Flow cytometry of LNs and tumor after subcutaneous injection of DIR‐EVs and DIR‐DP‐EVs in E.G7‐OVA tumor bearing mice. (G) Schematic illustration of the LNs and tumor targeting mechanism for DP‐EVs. All values presented in this figure are expressed as the mean ± s.d. Student's t ‐test was utilized for two‐group comparisons. * p < 0.05, ** p < 0.01. N = 3.

Article Snippet: The E.G7‐OVA and 4T1 cell line was obtained from the American Type Culture Collection.

Techniques: In Vivo, Fluorescence, Imaging, Saline, Labeling, Injection, Flow Cytometry

Journal: MedComm

Article Title: An Engineered Extracellular Vesicle With Enhanced Tumor and Lymph Nodes Targeting as siRNA Delivery System for Robust Tumor Immunotherapy

doi: 10.1002/mco2.70673

Figure Lengend Snippet: DP‐EVs can be effectively internalized by multiple cells and facilitated BMDCs migration and maturation. (A) Schematic illustration of DP‐EVs endocytosed by BMDCs. (B) Uptake efficiency of DIO‐EVs and DP‐EVs in E.G7‐OVA, BMDCs, and BMDMs. Scale bars, 100 µm. (C) Representative colocalization images in BMDCs of three uptake pathway markers with EVs and DP‐EVs, CT‐B, a marker of the caveolin pathway; transferrin, a marker of the clathrin pathway; and Dextran Texas, a marker of the macropinocytosis pathway. Scale bars, 20 µm. (D) Uptake efficiency of DIO‐EVs and DIO‐DP‐EVs in BMDCs pretreated with three inhibitors, amiloride (20 µM, an inhibitor of the macropinocytosis pathway); chlorpromazine (5 µM, an inhibitor of the clathrin‐mediated endocytosis pathway); and genistein (30 µM, an inhibitor of the caveolin‐mediated endocytosis pathway). (E) Representative colocalization images at 24 h after incubation in BMDCs of early endosomes, late endosomes, and lysosomes with EVs and DP‐EVs. Scale bars, 10 µm. (F) The percentages of mature DCs after different treatments. (G) The proportion of M1/M2 after different treatment. (H) The expression of CCR7 after different treatments detected by qPCR. (I) The expression of CCR7, AKT, p‐AKT, PI3K, p‐PI3K, JNK, p‐JNK, MAPK, p‐MAPK, ERK, p‐ERK, p65, and p‐p65 proteins in BMDCs after different treatment. (J) Schematic illustration of DC migration caused by DP‐EVs. (K) The percentages of migrated DCs after different treatments. All values presented in this figure are expressed as the mean ± s.d. Statistical analysis was performed using one‐way ANOVA with Dunnett's multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. N = 3.

Article Snippet: The E.G7‐OVA and 4T1 cell line was obtained from the American Type Culture Collection.

Techniques: Migration, Marker, Incubation, Expressing, Comparison

Journal: MedComm

Article Title: An Engineered Extracellular Vesicle With Enhanced Tumor and Lymph Nodes Targeting as siRNA Delivery System for Robust Tumor Immunotherapy

doi: 10.1002/mco2.70673

Figure Lengend Snippet: DP‐EVs delivered siRNA to downregulate PD‐L1 both in E.G7‐OVA and BMDCs. (A) Schematic illustration of DP‐EVs loading siRNA. Scale bars = 100 nm. (B) TEM images DP‐EVs and DP‐EVs/siRNA. (C) Size detection by NTA of DP‐EVs and DP‐EVs/siRNA. (D) Zeta potential detection through NTA of DP‐EVs and DP‐EVs/siRNA. (E) RNase A protection assay of EVs/siRNA and DP‐EVs/siRNA. (F) Uptake efficiency of DIO‐EVs/cy3‐siRNA and DIO‐DP‐EVs/cy3‐siRNA. (G) Representative colocalization images at 6 and 24 h after incubation in BMDCs of early endosomes, late endosomes, and lysosomes with DP‐EVs/cy3‐siRNA. (H) Detection of PD‐L1 expression at the mRNA level in E.G7‐OVA and BMDCs by qPCR. (I and J) Detection of PD‐L1 expression at the protein level in E.G7‐OVA and BMDCs by western blot. All values presented in this figure are expressed as the mean ± s.d. Statistical analysis was performed using one‐way ANOVA with Dunnett's multiple comparison test. *** p < 0.001, **** p < 0.0001. N = 3.

Article Snippet: The E.G7‐OVA and 4T1 cell line was obtained from the American Type Culture Collection.

Techniques: Zeta Potential Analyzer, Incubation, Expressing, Western Blot, Comparison

Journal: MedComm

Article Title: An Engineered Extracellular Vesicle With Enhanced Tumor and Lymph Nodes Targeting as siRNA Delivery System for Robust Tumor Immunotherapy

doi: 10.1002/mco2.70673

Figure Lengend Snippet: Antitumor efficacy of DP‐EVs/siPD‐L1 in E.G7‐OVA subcutaneous tumor mouse model. (A) The timeline process of the experimental scheme. (B) Individual tumor growth curves for E.G7‐OVA tumors in mice after different treatments ( n = 7). (C) Average tumor growth curves for E.G7‐OVA tumors in mice after different treatments ( n = 7). (D) Survival of mice with E.G7‐OVA tumors after various treatments ( n = 7). (E and F) TUNEL, CD31, and Ki67 staining of E.G7‐OVA tumors after various treatments ( n = 3). All values presented in this figure are expressed as the mean ± s.d. Statistical analysis was performed using one‐way ANOVA with Dunnett's multiple comparison test. Survival curves were obtained using the Kaplan–Meier method and compared by the log‐rank test. * p < 0.05, *** p < 0.001.

Article Snippet: The E.G7‐OVA and 4T1 cell line was obtained from the American Type Culture Collection.

Techniques: TUNEL Assay, Staining, Comparison

Journal: MedComm

Article Title: An Engineered Extracellular Vesicle With Enhanced Tumor and Lymph Nodes Targeting as siRNA Delivery System for Robust Tumor Immunotherapy

doi: 10.1002/mco2.70673

Figure Lengend Snippet: Detection of TME reprogramming and immune response activation after different treatments. (A) The timeline process of the experimental scheme. (B) t‐Distributed stochastic neighbor embedding (t‐SNE) results of DCs, macrophages, T cells, and CD3 − cells in E.G7‐OVA tumors after different treatments. (C) t‐SNE results of T cells in lymph nodes after different treatments. (D–K) Flow cytometry detection of DCs, macrophages, T cells, and CD3 − cells in E.G7‐OVA tumors’ microenvironments after different treatments. (L) Schematic illustration of TME variation after DP‐EVs/siPD‐L1 administration. (M and N) Flow cytometry detection of T cells in lymph nodes after different treatments. All values presented in this figure are expressed as the mean ± s.d. Statistical analysis was performed using one‐way ANOVA with Dunnett's multiple comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. N = 3.

Article Snippet: The E.G7‐OVA and 4T1 cell line was obtained from the American Type Culture Collection.

Techniques: Activation Assay, Flow Cytometry, Comparison

Journal: bioRxiv

Article Title: Targeting phosphodiesterase 10A disrupts MAPK signaling pathways in the tumor microenvironment to unleash antitumor immunity

doi: 10.64898/2026.02.26.708383

Figure Lengend Snippet: (A-C) Detection of ICD markers. Tumor cells were treated with DMSO or ADT-030 for 24 hours. Cell surface expression of CRT was detected by FACS, and MFI values are summarized in (A) as mean ± SEM. Extracellular ATP and HMGB1 levels from the indicated cells were measured and results shown as mean +/- SEM are summarized in (B) and (C), respectively. (D) ADT-030-treated tumor cells promote DC maturation. The schema depicts the experimental setup in which BMDCs were co-cultured with ADT-030 or DMSO pre-treated 4T1.GFP cells for 48 hrs. Uptake of the GFP signal by DCs and DC activation markers (MHC-II, CD80 and CD86) were assessed by FACS. MFIs are summarized as mean ± SEM in bar graphs. (E, F) cDC1-dependent priming of tumor-specific CD8⁺ T cells. The schema depicts the experimental procedures (E). EG7.OVA tumor cells were s.c. implanted to the flanks of WT or Batf3KO mice. Mice with palpable tumors were treated with vehicle or ADT-030. Peripheral blood was collected 10 days after treatment to detect OVA-specific CD8+ T cells by OVA-Kb tetramer. Representative dot plots are shown in (F). Numbers of the gated region demark percent of tetramer-positive cells in CD8+ T cell population. Results shown as mean +/- SEM are summarized in the bar graph. (G) ADT-030 efficacy in mice diminishes in the absence of cDC1. As depicted in the schema, EL4 or KPC cells were s.c. implanted to WT or Batf3KO mice. Mice with palpable tumors were treated daily with vehicle or ADT-030 daily till the endpoint. Tumor growth curves are shown as mean ± SEM, with the number of mice per group indicated. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Murine tumor cell lines 4T1, A20, B16F10, CT26, EL4, EG7.OVA and MC38 were purchased from ATCC.

Techniques: Expressing, Cell Culture, Activation Assay

Journal: bioRxiv

Article Title: Targeting phosphodiesterase 10A disrupts MAPK signaling pathways in the tumor microenvironment to unleash antitumor immunity

doi: 10.64898/2026.02.26.708383

Figure Lengend Snippet: (A) ADT-030 exhibits curative antitumor effect in EG7.OVA tumor model. Following the workflow depicted in the schema, WT C57BL/6 mice were implanted s.c. with EG7.OVA tumor cells. Mice with established tumors (50-100mm2) were treated with ADT-030 for 12 consecutive days. 3 mice that achieved complete tumor regression were rechallenged with EG7.OVA tumor cells. Naïve mice inoculated with EG7.OVA tumors at the same time servied as controls. (B) ADT-030 antitumor efficacy is compromised in Batf3KO mice. Following the workflow depicted in the schema, EG7.OVA cells were s.c. implanted to WT or Batf3KO mice. Mice with established tumors were treated daily with vehicle or ADT-030 daily till the endpoint. Tumor growth curves are shown as mean ± SEM, with the number of mice per group indicated. *, P < 0.05.

Article Snippet: Murine tumor cell lines 4T1, A20, B16F10, CT26, EL4, EG7.OVA and MC38 were purchased from ATCC.

Techniques: