Journal: Neoplasia (New York, N.Y.)

Article Title: Assessing axitinib-induced differential responses in tumor vascularization and oxygenation with combined optoacoustic angiography and diffuse optical spectroscopy

doi: 10.1016/j.neo.2026.101303

Figure Lengend Snippet: Axitinib inhibits Colo320 growth. ( a ) The experimental timeline: time points of axitinib administration are indicated by black triangles, time points of OA and DOS investigation - by white triangles, time point of morphological study – by red triangle. ( b ) Dynamics of the Colo320 tumor volumes during treatment with axitinib. Individual values, 25-75 percentiles, medians, minimum and maximum of the data set (n = 6). *, p < 0.05; **, p < 0.01 for treated versus control group (Wilcoxon test). # , p < 0.05 for current values versus initial level (Mann-Whitney test).

Article Snippet: All the experiments were conducted on subcutaneously implanted xenograft model based on the Colo320 colon adenocarcinoma cell line (Colo320DM, ATCC No CCL-220).

Techniques: Control, MANN-WHITNEY

Journal: Neoplasia (New York, N.Y.)

Article Title: Assessing axitinib-induced differential responses in tumor vascularization and oxygenation with combined optoacoustic angiography and diffuse optical spectroscopy

doi: 10.1016/j.neo.2026.101303

Figure Lengend Snippet: Axitinib-induced reduction of tumor vascularity. ( a ) Examples of OA images of Colo320 vasculature before and after treatment with axitinib. Bar is 3 mm. Dashed lines contour the tumor zones. ( b ) Volumetric vessel fraction of the Colo320 tumors during treatment with axitinib. ( c ) The corresponding projected vessel area. Individual values and M ± SD (n = 6). *, p < 0.05; **, p < 0.01 for treated versus control group (unpaired t-test). # , p < 0.05; # # , p < 0.01 for current values versus initial level (paired t-test).

Article Snippet: All the experiments were conducted on subcutaneously implanted xenograft model based on the Colo320 colon adenocarcinoma cell line (Colo320DM, ATCC No CCL-220).

Techniques: Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Assessing axitinib-induced differential responses in tumor vascularization and oxygenation with combined optoacoustic angiography and diffuse optical spectroscopy

doi: 10.1016/j.neo.2026.101303

Figure Lengend Snippet: Axitinib-induced decreases of CD31 positive blood vessels. ( a ) Examples of microimages from the tumor sections (scale bar 100 µm). ( b ) The numbers of CD31+ microvessels in the untreated and axitinib-treated Colo320 tumors after immunohistochemical staining for CD31. Individual values and M ± SD. ***, p < 0.001 for treated versus control group (unpaired t-test). ( c ), The values of CD31+ microvessels versus vascular fraction in the treated and untreated tumors.

Article Snippet: All the experiments were conducted on subcutaneously implanted xenograft model based on the Colo320 colon adenocarcinoma cell line (Colo320DM, ATCC No CCL-220).

Techniques: Immunohistochemical staining, Staining, Control

Journal: Neoplasia (New York, N.Y.)

Article Title: Assessing axitinib-induced differential responses in tumor vascularization and oxygenation with combined optoacoustic angiography and diffuse optical spectroscopy

doi: 10.1016/j.neo.2026.101303

Figure Lengend Snippet: Axitinib-induced increase of pimonidazole-positive areas. ( a ) Examples of LSM images taken from the tumor sections. Bar 2 mm. ( b ) Relative hypoxic fraction (RHF) of the untreated and axitinib-treated Colo320 tumors after immunofluorescent staining for hypoxia with pimonidazole. Individual values and M ± SEM. *, p < 0.05 for treated versus control group (unpaired t-test). ( c ) The RHF values versus StO 2 .

Article Snippet: All the experiments were conducted on subcutaneously implanted xenograft model based on the Colo320 colon adenocarcinoma cell line (Colo320DM, ATCC No CCL-220).

Techniques: Staining, Control

Journal: iScience

Article Title: Targeting USP14 enhances immunotherapy response by reprogramming tumor-associated macrophages in colon cancer

doi: 10.1016/j.isci.2026.115362

Figure Lengend Snippet: Targeting USP14 inhibits tumor growth and induces local anti-tumor immunity in vivo (A) Representative images of MC38 tumors harvested on day 21 post-inoculation from mice treated with IU1 (20 mg/kg, i.p., on days 6, 9, 12, and 15) or vehicle control. (B) Statistics of MC38 tumor growth rate following IU1 or vehicle treatment in vivo . The intraperitoneal injection dose of IU1 was 20 mg/kg, administered on days 6, 9, 12, and 15. Data are presented as the mean ± SEM ( n = 6 per group). (C) Spider diagram of the tumor volume growth in each mouse from the IU1 group and the PBS group. (D) Gating strategy for the detection of the TAMs by flow cytometry. We first obtained live cells, and then identified cells that were positive for CD45, CD11b, F4-80, and CD206 as M2 macrophages. (E–P) Proportions of neutrophil (E), M2 macrophage (F), M1 macrophage (G), MDSC (H), activated DCs (I), CD4 T cell (J), Treg cells (K), CD8 T cell (L), IFN-γ + CD8 T cell (M), precursor exhausted T cells (TCF-1 + ) (N), effective CD8 T cells (PD-1 + ) (O) and activated CD8 T cell (CD69 + ) (P) in the TME of the IU1 group and the control group by using flow cytometry. (Q–U) Cytokines IFN-γ (Q), TNF-α (R), IL-2 (S), IL-10 (T), and IL-12 (U) in the TME of each group were detected by Mul-Analyte Flow Assay Kit. (V) Schematic illustration of the proposed mechanism of action of IU1 in reprogramming the tumor microenvironment. Data are presented as the mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, and ns: not significant.

Article Snippet: Mouse colon cancer cells MC38 , Procell , CL-0972.

Techniques: In Vivo, Control, Injection, Flow Cytometry

Journal: iScience

Article Title: Targeting USP14 enhances immunotherapy response by reprogramming tumor-associated macrophages in colon cancer

doi: 10.1016/j.isci.2026.115362

Figure Lengend Snippet: Identification of the mechanism underlying IU1-mediated reprogramming of M2 macrophages (A and B) Verification of the efficiency of i.v. injection of clodronate liposome (Clo) in depleting TAMs of the blood (A) and the TME (B). (C and D) Statistics of tumor size monitored for 21 days in MC38 tumor-bearing mice after various indicated treatments. (E) Statistic of the percentage of CTLs in the TME after the indicated treatment. (F) Volcano plots of the differentially expressed genes between the PBS and the IU1 group. Red dots show significantly up-regulated genes in the IU1 group, and green dots show significantly down-regulated genes. (G) Heatmap illustrates the differentially expressed M1-and M2-related genes in TAMs in the IU1 group and the PBS group based on RNA sequencing results. (H) KEGG analysis identifies the 17 most enriched pathways based on the differentially expressed genes of the two groups. (I) Western blotting of p -JNK, p -ERK, p-p38, and GAPDH in IL-4/13-BMDM M2 cells treated with IU1 at the indicated time points. (J) RT-PCR to verify the typical M1/M2 polarization-related genes in M2 macrophages after treatment with IU1. (K) Flow cytometry analysis of CD206 expression on the IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, IU1 (10 μM) stimulation, IU1 (10 μM) stimulation in the presence of inhibitors of p38 (SB203580, 10 μM), JNK (SP600125, 10 μM), Erk1/2 (U0126-EtOH, 10 μM). Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparison test. Data are presented as the mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, and ns: not significant.

Article Snippet: Mouse colon cancer cells MC38 , Procell , CL-0972.

Techniques: Injection, RNA Sequencing, Western Blot, Reverse Transcription Polymerase Chain Reaction, Flow Cytometry, Expressing, Comparison

Journal: iScience

Article Title: Targeting USP14 enhances immunotherapy response by reprogramming tumor-associated macrophages in colon cancer

doi: 10.1016/j.isci.2026.115362

Figure Lengend Snippet: Combined blockade of USP14 and PD-1 exerts a synergistic anti-tumor effect in vivo (A) Gating strategy for distinguishing tumor cells, DC cells, and macrophages. (B) Flow cytometry was used to analyze the differences in PD-L1 expression in tumor cells, DC cells, and macrophages in the indicated treatment groups. (C) Flow cytometry was used to statistically analyze the differences in PD-L1 expression in tumor cells, DC cells, and macrophages in the specified groups. (D) Treatment schedule of the combination of IU1 and PD-1 antibody for the MC38 tumor-bearing mice. The intraperitoneal injection dose of IU1 was 20 mg/kg, administered on days 6, 9, 12, and 15. The intraperitoneal injection dose of anti-PD-1 antibody was 7.5 mg/kg, administered on days 7, 9, 11, and 13. (E and F) Tumor growth was monitored for 21 days in MC38 tumor-bearing mice after various indicated treatments. (G) Kaplan-Meier survival plot shows the survival of mice after the indicated treatments ( n = 10). (H–I) Multicolor immunofluorescence detection of M2 macrophage (H) and CTLs (I) in the TME of the indicated treatment group. Scale bar is 100 μm. Data are presented as the mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, and ns: not significant.

Article Snippet: Mouse colon cancer cells MC38 , Procell , CL-0972.

Techniques: In Vivo, Flow Cytometry, Expressing, Injection, Immunofluorescence

Journal: Biomaterials Research

Article Title: A Peptide Inhibitor of Lymphocyte Activation Gene-3 Interaction with Fibrinogen-like Protein 1 Synergizes with Programmed Death-Ligand 1 Blockade to Restore T Cell Activity and Inhibit Tumor Growth

doi: 10.34133/bmr.0364

Figure Lengend Snippet: Combined treatment with LAG3pep-2 and anti-programmed death-ligand 1 (anti-PD-L1) antibody restores T cell activity against tumor cells. (A) Experimental schemes. CD8+ T cells were isolated from the spleen of MC38 tumor-bearing mice and incubated for 48 h with anti-CD3/CD28 beads (activation) and interleukin-2 (IL-2)/interleukin-15 (IL-15) (proliferation). The activated T cells were co-cultured with MC38 cells in the absence or presence of LAG3pep-2 and anti-mouse PD-L1 antibody alone or in combination. Created with BioRender.com . (B) Activated CD8+ T cells were stained with carboxyfluorescein succinimidyl ester (CFSE) dye and co-cultured with tumor cells for 24 h. The population of CD3+/CFSE− cells was measured. (C to E) After co-culturing for 24 h, the culture medium was collected, and the percentage of cell death (lactate dehydrogenase [LDH] release) (C) and the concentrations of interferon-γ (IFN-γ) (D) and granzyme B (E) were measured. Data are presented as the mean ± SD of 3 independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant by one-way analysis of variance (ANOVA).

Article Snippet: HEK 293T (human embryonic kidney) cells, Jurkat T (human acute T cell leukemia) cells, THP-1 (acute monocytic leukemia) cells, HepG2 (human liver cancer) cells, and MC38 (C57BL/6 murine colon tumor) cells were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Activity Assay, Isolation, Incubation, Activation Assay, Cell Culture, Staining

Journal: Biomaterials Research

Article Title: A Peptide Inhibitor of Lymphocyte Activation Gene-3 Interaction with Fibrinogen-like Protein 1 Synergizes with Programmed Death-Ligand 1 Blockade to Restore T Cell Activity and Inhibit Tumor Growth

doi: 10.34133/bmr.0364

Figure Lengend Snippet: LAG3pep-2 synergizes with anti-programmed death-ligand 1 (anti-PD-L1) antibody to inhibit tumor growth and enhance anti-tumor immunity. (A) Experimental schemes. Mice bearing subcutaneous MC38 tumor were treated with intravenous (iv) administration of LAG3pep-2 and intraperitoneal (ip) administration of anti-mouse PD-L1 and anti-mouse lymphocyte activation gene-3 (LAG-3) antibodies (10 mg/kg body weight for single treatment and 5 mg/kg body weight for combined treatment). Created with BioRender.com . (B) Tumor volumes (mm 3 ) after treatments. Data are presented as mean ± SD ( n = 5/group). * P < 0.05 ** P < 0.01 from 2-way analysis of variance (ANOVA) including all treatment groups. (C) Tumor volumes (mm 3 ) of each treatment group after tumor inoculation. (D to G) Tumor tissue cell suspensions were prepared and gated for CD45+CD3+ lymphocytes. The populations of CD4+ T cells (D), CD8+ T cells (E), and FoxP3+ T cells (F) and the ratio of CD8+ T cells/FoxP3+ T cells (G) were measured. Data are presented as mean ± SD ( n = 4/group). * P < 0.05; *** P < 0.01; ns, not significant by one-way ANOVA.

Article Snippet: HEK 293T (human embryonic kidney) cells, Jurkat T (human acute T cell leukemia) cells, THP-1 (acute monocytic leukemia) cells, HepG2 (human liver cancer) cells, and MC38 (C57BL/6 murine colon tumor) cells were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Activation Assay

Journal: Biomaterials Research

Article Title: A Peptide Inhibitor of Lymphocyte Activation Gene-3 Interaction with Fibrinogen-like Protein 1 Synergizes with Programmed Death-Ligand 1 Blockade to Restore T Cell Activity and Inhibit Tumor Growth

doi: 10.34133/bmr.0364

Figure Lengend Snippet: LAG3pep-2 does not induce immunotoxicity in mice. (A) LAG3pep-2 was administered intraperitoneally to keyhole limpet hemocyanin (KLH)-challenged C57BL/6 mice at the indicated doses (0.01 to 100 mg/kg body weight). Cyclophosphamide (CPA) was used as a positive control. Mice were sacrificed, and spleen weights were measured on day 7 posttreatment. (B and C) Serum levels of total IgG2a (B) and total IgE (C). (D to F) The population of CD4+IFN-γ+ T cells (D), CD4+IL-4+ T cells (E), and CD4+IL-17A+ T cells (F) in the spleen. Data are presented as mean ± SD ( n = 5/group).

Article Snippet: HEK 293T (human embryonic kidney) cells, Jurkat T (human acute T cell leukemia) cells, THP-1 (acute monocytic leukemia) cells, HepG2 (human liver cancer) cells, and MC38 (C57BL/6 murine colon tumor) cells were obtained from the American Type Culture Collection (Manassas, VA).

Techniques: Positive Control

Journal: bioRxiv

Article Title: Lung-targeted cytokine-coding RNA-lipoplexes induce T and NK cell-mediated anti-tumor immune response

doi: 10.64898/2026.05.06.723126

Figure Lengend Snippet: ( A-D ) Luc–encoding RNA formulated as RNA-LPX was administered i.v. to non-tumor bearing (A, B; n=3) or CT26 metastases-bearing (C, D) BALB/c mice (n=3-9). ( A ) BLI of total body, ( B ) normalized organ signal from explanted organs. Data were analyzed by ordinary one-way ANOVA and Tukey’s test for multiple comparisons. ( C ) Kinetics of BLI of the lung signal in metastases-bearing mice, significance was determined by mixed-effects analysis with Geisser-Greenhouse correction and Tukey’s test for multiple comparisons. ( D ) Luc RNA, luc protein and CD31 (PECAM-1) were detected via RNAscope and immunohistochemistry on consecutive sections 1, 6 or 24 hours post injection, scale bar: 100 µm, tumor tissue is indicated with dashed line. ( E ) Cytokine quantification in lung after the indicated time points. ( F ) Cytokine fold increase in lung normalized to spleen at 6 hours after injection. ( G ) FDG positron emission tomography (PET) imaging 24 h after cytokine RNA mix injection into naïve mice, ( H ) ex vivo measurement of FDG uptake. Significance was determined using a two-tailed t-test for unpaired samples.

Article Snippet: Colon Carcinoma cell line CT26 was purchased from ATCC (CT26: CRL-2638, lot no. 58494154, female).

Techniques: RNAscope, Immunohistochemistry, Injection, Positron Emission Tomography, Imaging, Ex Vivo, Two Tailed Test

Journal: bioRxiv

Article Title: Lung-targeted cytokine-coding RNA-lipoplexes induce T and NK cell-mediated anti-tumor immune response

doi: 10.64898/2026.05.06.723126

Figure Lengend Snippet: ( A ) Experimental design: BALB/c mice (n=15 per group) were injected i.v. with CT26 tumor cells; subsequently, cytokine RNA mix or irrelevant RNA was administered i.v. as RNA-LPX twice per week for a total of 8 injections from d3 to d27. Survivor mice from the cytokine RNA mix-treated group (n=5) compared to naïve BALB/c mice (n=10) were re-challenged with CT26 tumor cells. ( B ) Survival after the initial i.v. CT26 tumor cell inoculation. ( C ) Survival after re-challenge. Significance was determined via Mantel-Cox logrank. ( D-F ) BALB/c mice (n=5 per group) were injected i.v. with CT26 tumor cells, RNA-LPX was administered i.v. at d3, d6 and d10 p.t.i., mice were sacrificed at d11, lungs were collected and analyzed via flow cytometry. Significance for pairwise comparisons was determined by unpaired two-tailed t-test. Flow cytometric analysis of ( D ) tumor burden, ( E ) CD8 + T and NK cells and ( F ) CD4 + Foxp3 + CD25 + T reg . ( G ) Functional analysis: after tumor cell inoculation, mice were treated with RNA-LPX at d3, d6, d10 and d13 p.t.i., mice were sacrificed at d14 for an ex vivo stimulation assay with PMA/Ionomycin ( H ) qRT-PCR was done to determine fold-change expression of cytokine RNA mix-treated normalized to irrelevant RNA LPX-treated lung samples (n=5 per group indicated in rows) for the indicated cytokines and chemokines.

Article Snippet: Colon Carcinoma cell line CT26 was purchased from ATCC (CT26: CRL-2638, lot no. 58494154, female).

Techniques: Injection, Flow Cytometry, Two Tailed Test, Functional Assay, Ex Vivo, Quantitative RT-PCR, Expressing

Journal: bioRxiv

Article Title: Lung-targeted cytokine-coding RNA-lipoplexes induce T and NK cell-mediated anti-tumor immune response

doi: 10.64898/2026.05.06.723126

Figure Lengend Snippet: Experimental design: BALB/c mice (n=5 per group) were injected i.v. with CT26 tumor cells; cytokine RNA mix or irrelevant RNA was administered i.v. at d3, d6 and d10 post tumor cell injection, mice were sacrificed at d11, lungs were collected and pooled at same ratios before sorting of CD45 + cells. CD45 + cells were subjected to scRNAseq. ( A,B ) Uniform manifold approximation and projection (UMAP) of 22 assigned clusters ( A ) and cell type frequencies ( B ) of CD45 + cells isolated from the lungs of cytokine RNA mix-treated versus irrelevant RNA-treated mice. ( C ) Effector function visualized as bubble plot. ( D ) Selected differentially expressed genes in cytokine RNA mix-treated versus irrelevant RNA-treated samples are shown as average log2 fold change. Note only significantly expressed values (p ≤ 0.05) are shown in colouring (blue = downregulated, red = upregulated), non-significant values are set to “0”/white.

Article Snippet: Colon Carcinoma cell line CT26 was purchased from ATCC (CT26: CRL-2638, lot no. 58494154, female).

Techniques: Injection, Isolation

Journal: bioRxiv

Article Title: Lung-targeted cytokine-coding RNA-lipoplexes induce T and NK cell-mediated anti-tumor immune response

doi: 10.64898/2026.05.06.723126

Figure Lengend Snippet: ( A ) Experimental design for ( B-C; CT26 tumor model) and ( E; CT26 B2M k.o. tumor model); n=15 BALB/c mice per group. Depletion or blocking antibody treatment was started 2 days prior to RNA-LPX treatment to ensure depletion before treatment start. ( B, C ) Survival according to termination criteria. ( D ) Experimental design and survival in CT26B2M k.o. or CT26gp70 k.o. tumor cells i.v. tumor model, n=15 mice per group. ( E ) BALB/c mice injected i.v. with CT26B2M k.o and depletion/neutralization antibodies were applied as shown in ( A ). Note that data in ( E ) were generated within the same experiment, irrelevant RNA + isotype mix as well as cytokine mix RNA + isotype mix refers to the same groups in all 3 plots. Survival was analyzed via Mantel-Cox logrank test. For ( B ) and ( C ), groups 1 and 2 were furthermore compared via logrank test with emphasis on early and late differences, ( B ) ## Logrank test with emphasis on late differences ((rho=0, lambda=1): group 1 vs 2: p=0,00235; ( C ) # Logrank test with emphasis on early differences (rho=1, lambda=0): group 1 vs 2: p=0,0378.

Article Snippet: Colon Carcinoma cell line CT26 was purchased from ATCC (CT26: CRL-2638, lot no. 58494154, female).

Techniques: Blocking Assay, Injection, Neutralization, Generated