Journal: FEBS Open Bio

Article Title: Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps

doi: 10.1002/2211-5463.13499

Figure Lengend Snippet: Liraglutide inhibits tumor progression through neutrophils. Measure blood cells in (A) LLC and (B) Hepa1‐6 cells bearing C57BL/6 mice using complete blood count. Concentrations of circulating (C) GM‐CSF, (D) VEGF‐A, (E) MMP‐9, (F) myeloperoxidase, and (G) elastase in mice were measured using ELISA. (H) Circulating dsDNA was purified and measured using an ultramicrospectrophotometer. Statistical significance was determined with t ‐test. N = 6 mice for every group. Nonparametric tests were utilized to assess statistical significance between different treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001. Error bars represent SD.

Article Snippet: Murine Lewis lung cancer cell LLC and liver cancer cell Hepa1‐6 were cultured in Dulbecco's Modified Eagle's Medium containing 10% FBS and 1% streptomycin–penicillin (10 000 U·mL −1 ) (all: Corning, New York, NY, USA) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Enzyme-linked Immunosorbent Assay, Purification

Journal: FEBS Open Bio

Article Title: Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps

doi: 10.1002/2211-5463.13499

Figure Lengend Snippet: Liraglutide reduced tumor‐infiltrated NETs. The LLC and Hepa1‐6 cells bearing C57BL/6 mice were treated using PBS or liraglutide, and neutrophils were enriched for in vitro study. Representative images of tumor‐derived NETs from (A) LLC and (C) Hepa1‐6 cells bearing C57BL/6 mice. Blue is nuclear stained by Hoechst 33342. Green is DNA stained by SYTOX Green. Quantification of tumor‐derived NETs from (B) LLC and (D) MC38 cells bearing C57BL/6 mice were conducted using a fluorescence microplate reader ( n = 3). The tumor infiltrated neutrophils from (E) LLC and (G) Hepa1‐6 cells bearing C57BL/6 mice were stained with total reactive oxygen species (ROS) assay kit, ROS production was analyzed by flow cytometry, and the green fluorescence of ROS was detected using FITC channel. Mean fluorescence intensity (MFI) of ROS in tumor‐derived neutrophils from (F) LLC and (H) MC38 cells bearing C57BL/6 mice were conducted using a fluorescence microplate reader ( n = 3). Nonparametric tests were utilized to assess statistical significance between different treatment groups. * P < 0.05, *** P < 0.001. Error bars represent SD. Scale bar: 400 μm (A, C).

Article Snippet: Murine Lewis lung cancer cell LLC and liver cancer cell Hepa1‐6 were cultured in Dulbecco's Modified Eagle's Medium containing 10% FBS and 1% streptomycin–penicillin (10 000 U·mL −1 ) (all: Corning, New York, NY, USA) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: In Vitro, Derivative Assay, Staining, Fluorescence, ROS Assay, Flow Cytometry

Journal: FEBS Open Bio

Article Title: Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps

doi: 10.1002/2211-5463.13499

Figure Lengend Snippet: Liraglutide enhanced the antitumor efficacy of PD‐1 blockade. Tumor volumes were monitored in (A) LLC or (B) Hepa1‐6 allograft mice treated by IgG isotype control (isotype), anti‐PD‐1 mAb (αPD‐1), liraglutide, or liraglutide and anti‐PD‐1 mAb (liraglutide+αPD‐1). (C) LLC and (D) Hepa1‐6 tumor volumes for the NETs‐removed C57BL/6 mice. NETs were removed using DNase I (5 mg·kg −1 ). (E) LLC and (F) Hepa1‐6 tumor volumes for the NETs‐removed C57BL/6 mice accepting isotype, αPD‐1, liraglutide, or liraglutide+αPD‐1 treatments. NETs were removed using DNase I (5 mg·kg −1 ). Tumor volume = 0.5 × length × width 2 . Statistical significance was determined with two‐way ANOVA tests. N = 12 mice for every group. Nonparametric tests were utilized to assess statistical significance between different treatment groups. ** P < 0.01, *** P < 0.001. Error bars represent SD.

Article Snippet: Murine Lewis lung cancer cell LLC and liver cancer cell Hepa1‐6 were cultured in Dulbecco's Modified Eagle's Medium containing 10% FBS and 1% streptomycin–penicillin (10 000 U·mL −1 ) (all: Corning, New York, NY, USA) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Control

Journal: FEBS Open Bio

Article Title: Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps

doi: 10.1002/2211-5463.13499

Figure Lengend Snippet: Long‐term immune responses induced by the combination therapy rejected rechallenge. The measurement of IFN‐γ concentration in the culture media of CD8 + T cells from (A, B) lymph node, (C, D) spleen, and (E, F) tumor tissues in LLC or Hepa1‐6 allograft mice after 7 days of treatment. N = 12 mice for every group. (G) LLC or (H) Hepa1‐6 allograft C57BL/6 mice were treated with STR. Mice pre‐treated with PBS, or the combination therapy received tumor rechallenge by administration of respective cell lines. N = 6 mice for every group. Tumor volume = 0.5 × length × width 2 . Statistical significance was determined with two‐tailed t ‐test. Nonparametric tests were utilized to assess statistical significance between different treatment groups. * P < 0.05, ** P < 0.01, *** P < 0.001. Error bars represent SD.

Article Snippet: Murine Lewis lung cancer cell LLC and liver cancer cell Hepa1‐6 were cultured in Dulbecco's Modified Eagle's Medium containing 10% FBS and 1% streptomycin–penicillin (10 000 U·mL −1 ) (all: Corning, New York, NY, USA) in a humidified incubator at 37 °C containing 5% CO 2 .

Techniques: Concentration Assay, Two Tailed Test

Journal: Cancer Research

Article Title: The circRNA cEMSY Induces Immunogenic Cell Death and Boosts Immunotherapy Efficacy in Lung Adenocarcinoma

doi: 10.1158/0008-5472.CAN-24-1484

Figure Lengend Snippet: cEMSY expression level correlates with an enhanced immunotherapy response. A, Illustrative images of 65 lung adenocarcinoma (LUAD) tissues showing cEMSY (magenta), CD8 + T cells (green), CD11c + DCs (white), and DAPI (blue). Samples were categorized based on cEMSY fluorescence into cEMSY high (above the mean intensity) and cEMSY low (below the mean intensity). B, Analysis of the proportion of intratumor CD11c + DCs and CD8 + T cells within DAPI + cells across the two cEMSY stratifications. Data presented as median ± quartiles for groups cEMSY low ( n = 37) and cEMSY high ( n = 28). C, The Kaplan–Meier curve for the disease-free survival (DFS) of patients with LUAD, who were divided into two groups according to the mean expression of cEMSY. D, Heatmap showing Spearman correlations between CD11c + DCs and CD8 + T-cell infiltration and cEMSY signature gene set variation analysis score of indicated gene sets in each TCGA cohort. ACC, adrenocortical carcinoma; BLCA, bladder urothelial Carcinoma; BRCA, breast invasive carcinoma; CESC, cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL, cholangiocarcinoma; COAD, colon adenocarcinoma; DLBC, lymphoid neoplasm diffuse large B-cell lymphoma; ESCA, esophageal carcinoma; GBM, glioblastoma multiforme; HNSC, head and neck squamous cell carcinoma; KICH, kidney chromophobe; KIRC, kidney renal clear cell carcinoma; KIRP, kidney renal papillary cell carcinoma; LGG, lower grade glioma; LIHC, liver hepatocellular carcinoma; LUSC, lung squamous cell carcinoma; MESO, mesothelioma; OV, ovarian serous cystadenocarcinoma; PAAD, pancreatic adenocarcinoma; PCPG, pheochromocytoma and paraganglioma; PRAD, prostate adenocarcinoma; READ, rectum adenocarcinoma; SARC, sarcoma; SKCM, skin cutaneous melanoma; STAD, stomach adenocarcinoma; TGCT, tescticular germ cell tumors; THCA, thyroid carcinoma; THYM, thymoma; UCEC, uterine corpus endometrial carcinoma; UCS, uterine carcinosarcoma; UVM, uveal melanoma. E, cEMSY signature expression level of the responders (R) and nonresponders (NR) of two representative LUAD ICB cohorts. F, Progression-free survival (PFS) of ICB cohorts based on the level of cEMSY signature. Statistics were applied using Spearman rank correlation, Student t test, or log-rank test.

Article Snippet: Human bronchial epithelial (HBE) cells and human lung adenocarcinoma cell lines (H358, A549, H23, H1975, PC9, A427, and H373), as well as murine lung adenocarcinoma cell lines (LLC1 and LA795), were obtained from the China Center for Type Culture Collection.

Techniques: Expressing, Fluorescence