Journal: Precision Clinical Medicine

Article Title: Agrimol B inhibits pancreatic ductal adenocarcinoma by induction of lethal mitophagy through decreasing mitochondrial transcription termination factor 3

doi: 10.1093/pcmedi/pbag009

Figure Lengend Snippet: Agrimol B sensitizes PDAC cells to first-line chemotherapy drugs. (A) Schematic overview of PDAC PDO establishment and drug assessment. (B, C) Brightfield images of organoids treated with the indicated concentrations of Agrimol B. Scale bars, 10 μm. (D) Chemical structures of nab-paclitaxel (Paclitaxel), irinotecan, 5-fluorouracil, oxaliplatin, and gemcitabine. (E–H) Brightfield images of organoids treated with or without Agrimol B in the presence or absence of nab-paclitaxel, irinotecan, 5-fluorouracil, oxaliplatin, or gemcitabine. Scale bars, 10 μm.

Article Snippet: Hydroxychloroquine (HCQ) (HY-W031727), wortmannin (WORT) (HY-10197), Mdivi-1 (HY-15886), N-acetylcysteine (HY-B0215), Nab-Paclitaxel (HY-P99974), irinotecan (HY-16562), 5-fluorouracil (5-FU) (HY-107856), and oxaliplatin (HY-17371) were purchased from MedChem Express.

Techniques:

Journal: ImmunoTargets and Therapy

Article Title: Irinotecan Modulates Immune Checkpoints in Neuroblastoma

doi: 10.2147/ITT.S598469

Figure Lengend Snippet: Irinotecan and IFN-ɣ modulate immune checkpoint expression in neuroblastoma cells. Determination of sub-lethal irinotecan concentrations ( A and B ). SK-N-BE(2) ( A ) and SH-SY5Y ( B ) cells were treated with increasing concentrations of irinotecan, and cell growth was monitored using the IncuCyte live-cell imaging system. Sub-lethal dose was defined as ≥70% viability after 48 hours. Surface expression of immune-related markers following treatment ( C - J ). SK-N-BE(2) ( C – F ) or SH-SY5Y ( G – J ) cells were treated with irinotecan (1 µM), IFN-γ (20 ng/mL), or their combination for the indicated time points. Expression of PD-L1 ( C,G ), MHC class I ( D,H ), CD47 ( E,I ), and GD2 ( F and J ) was assessed by flow cytometry and presented as mean fluorescence intensity (MFI). Treatment groups: control (purple), irinotecan (blue), IFN-γ (green), irinotecan + IFN-γ (Orange). Data are presented as mean ± SEM. *p < 0.05; **p < 0.01.

Article Snippet: Irinotecan (Irinotecan HCl, Pfizer) was added directly to the media at noted concentrations.

Techniques: Expressing, Live Cell Imaging, Flow Cytometry, Fluorescence, Control

Journal: ImmunoTargets and Therapy

Article Title: Irinotecan Modulates Immune Checkpoints in Neuroblastoma

doi: 10.2147/ITT.S598469

Figure Lengend Snippet: Irinotecan reduces macrophage-mediated phagocytosis, which is restored by CD47 blockade. GFP-expressing neuroblastoma cells (SK-N-BE(2) or SH-SY5Y) were pre-treated with irinotecan (1 µM) for 24 hours and co-cultured with human macrophages derived from PBMCs at a 1:1 ratio. Anti-CD47 antibody (αCD47, 10 µg/mL) was added where indicated. ( A ) Phagocytosis efficiency, defined as the percentage of CD11b⁺GFP⁺ macrophages, normalized to untreated control (set as 100%). ( B ) Quantification of live tumor cells, defined as GFP⁺ viable cells by flow cytometry. ( C ) Phagocytic activity, assessed as GFP mean fluorescence intensity (MFI) within CD11b⁺GFP⁺ macrophages. Treatment groups: control (purple), irinotecan (blue), αCD47 (green), irinotecan + αCD47 (Orange). Data are presented as mean ± SEM.*p < 0.05; **p < 0.01.

Article Snippet: Irinotecan (Irinotecan HCl, Pfizer) was added directly to the media at noted concentrations.

Techniques: Expressing, Cell Culture, Derivative Assay, Control, Flow Cytometry, Activity Assay, Fluorescence

Journal: ImmunoTargets and Therapy

Article Title: Irinotecan Modulates Immune Checkpoints in Neuroblastoma

doi: 10.2147/ITT.S598469

Figure Lengend Snippet: Irinotecan pre-treatment enhances T cell–mediated cytotoxicity against neuroblastoma cells. Activated T cells were induced from human PBMCs by CD3 and CD28 co-stimulation. NBL cells, SK-N-BE(2) and SH-SY5Y, were pre-treated with 1µM of Irinotecan for 24hrs. NBL and T cells were co-cultured for 24hrs. Live NBL cells were assessed by FACS. Data are presented as the change in the percentage of tumor cell death compared to untreated control cells. Representative data from SK-N-BE(2) cells are shown. Treatment groups: Control (dot), T cells (blue). Data are shown as mean ± SEM. **p < 0.01.

Article Snippet: Irinotecan (Irinotecan HCl, Pfizer) was added directly to the media at noted concentrations.

Techniques: Cell Culture, Control

Journal: ImmunoTargets and Therapy

Article Title: Irinotecan Modulates Immune Checkpoints in Neuroblastoma

doi: 10.2147/ITT.S598469

Figure Lengend Snippet: Soluble CD47 is detectable in neuroblastoma cell culture supernatants and patient plasma. ( A ) SH-SY5Y cells were treated with irinotecan (1 µM) and/or IFN-γ (20 ng/mL) for 48 hours. CD47 levels in cell culture supernatants were measured by ELISA. ( B ) Plasma levels of soluble CD47 in neuroblastoma patients (n=5) were measured by ELISA. Samples were obtained at indicated clinical time points (pre-treatment and post-treatment where available). Patient 1 (light blue), patient 2 (purple), patient 3 (pink), patient 4 (green), patient 5 (dark blue). Data are presented as mean ± SEM. **p < 0.01.

Article Snippet: Irinotecan (Irinotecan HCl, Pfizer) was added directly to the media at noted concentrations.

Techniques: Cell Culture, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: ImmunoTargets and Therapy

Article Title: Irinotecan Modulates Immune Checkpoints in Neuroblastoma

doi: 10.2147/ITT.S598469

Figure Lengend Snippet: Effects of irinotecan and PBMCs on tumor burden in a humanized neuroblastoma mouse model. NSG mice were intraperitoneally injected with luciferase-expressing SH-SY5Y cells. On day 9, mice received total body irradiation (2 Gy) and were randomized into treatment groups: control, PBMCs alone, irinotecan alone, or irinotecan + PBMCs. PBMCs (2 × 10 6 cells) were administered intraperitoneally, and irinotecan (400 µg/mouse) intravenously. ( A ) Representative bioluminescence imaging (BLI) of tumor burden.( B ) Quantification of BLI signal over time. ( C ) Tumor burden at day 15. Treatment groups: control (purple), irinotecan (green), PBMCs (blue), irinotecan + PBMCs (Orange).Data are presented as mean ± SEM. *p < 0.05.

Article Snippet: Irinotecan (Irinotecan HCl, Pfizer) was added directly to the media at noted concentrations.

Techniques: Injection, Luciferase, Expressing, Irradiation, Control, Imaging

Journal: ImmunoTargets and Therapy

Article Title: Irinotecan Modulates Immune Checkpoints in Neuroblastoma

doi: 10.2147/ITT.S598469

Figure Lengend Snippet: Combined irinotecan and PBMC treatment is associated with increased human T-cell representation in spleen. Neuroblastoma tumors were induced by intra-peritoneal injection of NBL SH-SY5Y-LUC cells. On day 10 mice were treated with either PBMCs or Irinotecan or a combination of both. At day 31 post-treatment, spleens were harvested and analyzed by flow cytometry. ( A ) Spleen weight. ( B ) Percentage of human leukocytes (hCD45⁺) among total splenocytes. ( C ) Percentage of human T cells (hCD3⁺) among human leukocytes. ( D ) Percentage of CD8⁺ T cells among human T cells. Treatment groups: control (purple), irinotecan (blue), PBMCs (green), irinotecan + PBMCs (Orange). Data are presented as mean ± SEM.

Article Snippet: Irinotecan (Irinotecan HCl, Pfizer) was added directly to the media at noted concentrations.

Techniques: Injection, Flow Cytometry, Control