Journal: NPJ Precision Oncology

Article Title: Single-organoid analysis reveals clinically relevant treatment-resistant and invasive subclones in pancreatic cancer

doi: 10.1038/s41698-023-00480-y

Figure Lengend Snippet: a Bar plot representation of the progression-free survival (PFS) in months per patient. b NDR results upon treatment with gemcitabine-paclitaxel highlighting the 3 distinct responses. Error bars indicate the standard deviation. c Representative brightfield/fluorescence (Cytotox Green) images of PDAC052, PDAC060 and PDAC082 indicating the presence of persistent PDAC organoid clones (black circle). One representative image was selected for visualization out of the two technical replicates. Scale bar=100 µm d Single organoid dose response based on cell death (green area/brightfield area) labeled as fraction affected and area (brightfield) of PDAC052, PDAC060 and PDAC082 treated with gemcitabine-paclitaxel (400 nM:80 nM). Dark grey region (Fraction affected <0.15) indicates resistant, middle grey region sensitive (Fraction affected 0.15-0.34) and light grey highly sensitive (Fraction affected >0.34) PDAC organoid clones. Bubble size correlates with the organoid area. e Relative fraction of resistant, sensitive and highly sensitive PDAC organoids for each patient treated with gemcitabine-paclitaxel (400 nM:80 nM). f Dynamic quantification of single-organoid responses treated with gemcitabine-paclitaxel (400 nM:80 nM) or FOLFIRINOX (20 µM 5-FU:0.0625 µM SN38:2.5 µM Oxaliplatin). The single organoid data was combined from 2 technical replicates.

Article Snippet: Cytotox Green (60 nM / well, Sartorius), Staurosporine (2 µM), 5-Fluorouracil (5-FU), SN38 (active metabolite Irinotecan), gemcitabine and paclitaxel (MedChemExpress) were dissolved in DMSO.

Techniques: Standard Deviation, Fluorescence, Clone Assay, Labeling

Journal: NPJ Precision Oncology

Article Title: Single-organoid analysis reveals clinically relevant treatment-resistant and invasive subclones in pancreatic cancer

doi: 10.1038/s41698-023-00480-y

Figure Lengend Snippet: a – c Spearman rank correlation of the % sensitive, % resistant and ratio % sensitive/resistant PDAC clones (gemcitabine-paclitaxel; 400 nM: 80 nM) with the PFS. d . Correlation of % ratio sensitive/resistant with the initial clinical response to gemcitabine-paclitaxel. Responses were based on available radiological CT protocols. Good= tumor regression and/or PFS > 10 months, Mixed= minor tumor regression, Bad= no tumor regression and/or fast disease progression. e NDR signatures indicating the NDR value (left y-axis) and % cell death (right y-axis) upon treatment with gemcitabine-paclitaxel (400 nM:80 nM) or FOLFIRINOX (4 µM 5-FU:0.0125 µM SN38:0.5 µM Oxaliplatin). f Representative CT-scans before and after treatment with gemcitabine/nab-paclitaxel or FOLFIRINOX (8 cycles). Red contours indicate the tumor margins. g Overview of the clinical characteristics of patient PDAC044, PDAC052, PDAC060, PDAC068 and PDAC087. h Comparison of the implemented readouts, suggesting a potential clinical application of combining the NDR readout with the developed single-organoid analysis.

Article Snippet: Cytotox Green (60 nM / well, Sartorius), Staurosporine (2 µM), 5-Fluorouracil (5-FU), SN38 (active metabolite Irinotecan), gemcitabine and paclitaxel (MedChemExpress) were dissolved in DMSO.

Techniques: Clone Assay, Comparison

Journal: Cell Reports Medicine

Article Title: Histone lysine demethylase 4 family proteins maintain the transcriptional program and adrenergic cellular state of MYCN-amplified neuroblastoma

doi: 10.1016/j.xcrm.2024.101468

Figure Lengend Snippet:

Article Snippet: Irinotecan , MedChem Express , HY-16562A.

Techniques: Recombinant, Modification, Synthesized, Saline, Transfection, Apoptosis Assay, Purification, Transgenic Assay, Control, Expressing, Plasmid Preparation, Software

Journal: Cancer Medicine

Article Title: Maritoclax Overcomes FBW7 Deficiency‐Driven Irinotecan Resistance in Colorectal Cancer by Targeting MCL1

doi: 10.1002/cam4.71419

Figure Lengend Snippet: Maritoclax enhanced the antitumor effect of irinotecan on FBW7 R465C‐overexpressed CRC in vivo. HA‐Vec (wild‐type group) or HA‐FBW7 R465C ‐overexpressed (R465C‐mutation group) HCT‐116 cells were subcutaneously injected into 5‐week‐old female BALB/c nude mice. Drugs were administered every 4 days after tumor volume reached 100 mm 3 . Nude mice in the wild‐type group were assigned to vehicle (DMSO) or irinotecan (35 mg/kg) treatment groups and those in R465C‐mutation group were divided into four groups that treated with vehicle (DMSO), irinotecan (35 mg/kg), Maritoclax (5 mg/kg), or irinotecan (35 mg/kg) plus Maritoclax (5 mg/kg), respectively. Nude mice were sacrificed when tumor volume reached 1500 mm 3 or diameter reached 20 mm. (A, D) Tumors were dissected and recorded by photographed. (B, E) Tumor volumes (length*width 2 /2) and (C, F) body weights of mice were recorded every other day during treatment. Western blot was performed to detect the protein level of MCL1 in tumor tissues from (G) the wild‐type and R465C‐mutation control groups; (H) the wild‐type control group and R465C‐mutation groups including control, irinotecan monotherapy, Maritoclax monotherapy, and combination therapy. Data are expressed as the mean ± SD ( n = 3, representing three independent experiments). p values were calculated by using Two‐way ANOVA or Student's t ‐test, the coloration of the p ‐value markers is consistent with the color scheme of each corresponding group. ns: p > 0.05, **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05 vs. the vehicle group; ### p < 0.001, ## p < 0.01, # p < 0.05 vs. the irinotecan group; ++ p < 0.01, + p < 0.05 vs. the Maritoclax group.

Article Snippet: Irinotecan (Cat# T6228; Targetmol), SN38 (Cat# T1703; Targetmol), Marinopyrrole A (Maritoclax, Cat# T11944 ; Targetmol) and MG132 (Cat# MB5137‐1; Meilunbio) were dissolved in dimethylsulfoxide (DMSO); puromycin (Cat# MB2005; Meilunbio) was dissolved in sterile deionized water.

Techniques: In Vivo, Mutagenesis, Injection, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Berberine Improves Irinotecan-Induced Intestinal Mucositis Without Impairing the Anti-colorectal Cancer Efficacy of Irinotecan by Inhibiting Bacterial β-glucuronidase

doi: 10.3389/fphar.2021.774560

Figure Lengend Snippet: Berberine ameliorated body weight loss, colon shortening, and histopathologic injury, in mice with CPT11-induced intestinal mucositis. (A) Changes in body weight in mice with CPT11-induced mucositis. The data are plotted as a percentage of the original body weight. Macroscopic observations (B) and the assessment of colon shortening (C) after berberine treatment. (D) Representative H&E-stained colon sections and histological scores. Scale bars correspond to 50 μm. Data are expressed as the mean ± SD ( n = 6 per group). ## p < 0.01 vs. the control group; *** p < 0.001 vs . CPT11-treated group.

Article Snippet: Irinotecan (CPT11) (C 33 H 45 N 4 O 9 ·Cl, CAS: 136572-09-3; molecular weight: 677.18) was purchased from Melone Pharmaceutical Co., Ltd (Dalian, China). β-glucuronidase (GUS, CAS: 9001-45-0, G7396-25kU) from Escherichia coli was obtained from Thermo Scientific Inc. (Waltham, MA, United States).

Techniques: Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Berberine Improves Irinotecan-Induced Intestinal Mucositis Without Impairing the Anti-colorectal Cancer Efficacy of Irinotecan by Inhibiting Bacterial β-glucuronidase

doi: 10.3389/fphar.2021.774560

Figure Lengend Snippet: Berberine inhibited the expression levels of pro-inflammatory mediators and promoted the mRNA expression levels of tight junction protein in vivo . (A) The effect of berberine on the expression of pro-inflammatory mediators (COX-2 and iNOS) in colonic tissues. Representative western blots and quantitative analysis of COX-2 and iNOS proteins. (B) mRNA expression levels of iNOS, IL-8, IL-1β, and TNF-α in colon tissue, as determined by qRT-PCR. (C) Representative Alcian blue (AB)/(Periodic Acid-Schiff stain) PAS-stained colonic tissue sections (Scale bars, 100 μm) and goblet cell number. (D) mRNA expression levels of Occludin, ZO-1, and Claudin-7, in colon tissue, as determined by qRT-PCR. All mRNA expression levels were normalized to β-Actin. Data are expressed as the mean ± SD ( n = 3 per group). ## p < 0.01, ### p < 0.001 vs. the control group; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CPT11 + berberine treated group.

Article Snippet: Irinotecan (CPT11) (C 33 H 45 N 4 O 9 ·Cl, CAS: 136572-09-3; molecular weight: 677.18) was purchased from Melone Pharmaceutical Co., Ltd (Dalian, China). β-glucuronidase (GUS, CAS: 9001-45-0, G7396-25kU) from Escherichia coli was obtained from Thermo Scientific Inc. (Waltham, MA, United States).

Techniques: Expressing, In Vivo, Western Blot, Quantitative RT-PCR, Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Berberine Improves Irinotecan-Induced Intestinal Mucositis Without Impairing the Anti-colorectal Cancer Efficacy of Irinotecan by Inhibiting Bacterial β-glucuronidase

doi: 10.3389/fphar.2021.774560

Figure Lengend Snippet: Berberine protected the integrity of the intestinal barrier in mice with mucositis. (A) Immunofluorescence staining of the colonic tissues for ZO-1 protein (green). Nuclei were stained with DAPI (blue). (B) The effect of berberine on the expression levels of tight junction proteins (ZO-1 and Claudin-7) in colonic tissues. Representative western blots and quantitative analysis of the proteins. (C) The fluorescence intensity of FITC-dextran was detected by fluorescence spectroscopy ( n = 4–5). The serum levels of LPS and DAO were determined by ELISA. Data are expressed as the mean ± SD ( n = 3 per group). ## p < 0.01, ### p < 0.001 vs. the control group; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CPT11 + berberine treated group.

Article Snippet: Irinotecan (CPT11) (C 33 H 45 N 4 O 9 ·Cl, CAS: 136572-09-3; molecular weight: 677.18) was purchased from Melone Pharmaceutical Co., Ltd (Dalian, China). β-glucuronidase (GUS, CAS: 9001-45-0, G7396-25kU) from Escherichia coli was obtained from Thermo Scientific Inc. (Waltham, MA, United States).

Techniques: Immunofluorescence, Staining, Expressing, Western Blot, Fluorescence, Spectroscopy, Enzyme-linked Immunosorbent Assay, Control

Journal: Frontiers in Pharmacology

Article Title: Berberine Improves Irinotecan-Induced Intestinal Mucositis Without Impairing the Anti-colorectal Cancer Efficacy of Irinotecan by Inhibiting Bacterial β-glucuronidase

doi: 10.3389/fphar.2021.774560

Figure Lengend Snippet: Effect of berberine on SN38-induced disruption of barrier function in vitro . (A) NCM460 cell monolayers were treated with berberine (50 μM), SN38 (250 nM) or SN38 (250 nM)) + Ber (50 μM) for 4, 12, 24, and 36 h, respectively; then, we measured trans-endothelial electrical resistance (TEER). (B) The flux of 4 kDa FITC-dextran was measured by fluorescence spectroscopy. (C) Representative images of scratch wound assays for NCM460 cell monolayers at 0 and 24 h after scratching. (D) The effect of berberine on the expression levels of tight junction proteins (ZO-1, Occludin, and Claudin-7) in NCM460 cells. (E) The expression of Occludin (red) was evaluated by immunofluorescence. Nuclei were stained with DAPI (blue). Data were expressed as the mean ± SD ( n = 3 per group). ## p < 0.01, ### p < 0.001 vs. the control group; * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CPT11 + berberine treated group.

Article Snippet: Irinotecan (CPT11) (C 33 H 45 N 4 O 9 ·Cl, CAS: 136572-09-3; molecular weight: 677.18) was purchased from Melone Pharmaceutical Co., Ltd (Dalian, China). β-glucuronidase (GUS, CAS: 9001-45-0, G7396-25kU) from Escherichia coli was obtained from Thermo Scientific Inc. (Waltham, MA, United States).

Techniques: Disruption, In Vitro, Fluorescence, Spectroscopy, Expressing, Immunofluorescence, Staining, Control

Journal: Frontiers in Pharmacology

Article Title: Berberine Improves Irinotecan-Induced Intestinal Mucositis Without Impairing the Anti-colorectal Cancer Efficacy of Irinotecan by Inhibiting Bacterial β-glucuronidase

doi: 10.3389/fphar.2021.774560

Figure Lengend Snippet: Berberine inhibited intestinal GUS activity. (A) Fecal pellets were collected to evaluate the GUS activity in mice after CPT11 and CPT11 + berberine treatment by 4-MUG assay. (B) The assessment of fecal pellets for GUS-producing bacteria was determined by culture with 4-MUG agar medium. (C) Mice were treated with CPT11 or CPT11 + berberine for 14 days; GUS activity was then imaged using the fluorescent substrate FDGlcU. Data were expressed as the mean ± SD ( n = 3 per group). ## p < 0.01, ### p < 0.001 vs. the control group; ** p < 0.01, *** p < 0.001 vs. CPT1 + berberine treated group.

Article Snippet: Irinotecan (CPT11) (C 33 H 45 N 4 O 9 ·Cl, CAS: 136572-09-3; molecular weight: 677.18) was purchased from Melone Pharmaceutical Co., Ltd (Dalian, China). β-glucuronidase (GUS, CAS: 9001-45-0, G7396-25kU) from Escherichia coli was obtained from Thermo Scientific Inc. (Waltham, MA, United States).

Techniques: Activity Assay, Mug Assay, Bacteria, Control

Journal: Frontiers in Pharmacology

Article Title: Berberine Improves Irinotecan-Induced Intestinal Mucositis Without Impairing the Anti-colorectal Cancer Efficacy of Irinotecan by Inhibiting Bacterial β-glucuronidase

doi: 10.3389/fphar.2021.774560

Figure Lengend Snippet: Berberine (50 mg/kg) improved the anti-tumor effects of CPT11 in a colon cancer xenograft model. (A) Image of colorectal tumor in each group. (B) Tumor weights were measured after animals were euthanized. (C) Tumor volumes were measured during the test period. (D) Changes in body weight were monitored throughout the study. (E) Images of hematoxylin and eosin staining in tumor tissues. Data are expressed as the mean ± SD ( n = 5 per group). ### p < 0.001 vs. the control group; *** p < 0.001 vs. CPT11 + berberine treated group.

Article Snippet: Irinotecan (CPT11) (C 33 H 45 N 4 O 9 ·Cl, CAS: 136572-09-3; molecular weight: 677.18) was purchased from Melone Pharmaceutical Co., Ltd (Dalian, China). β-glucuronidase (GUS, CAS: 9001-45-0, G7396-25kU) from Escherichia coli was obtained from Thermo Scientific Inc. (Waltham, MA, United States).

Techniques: Staining, Control