Journal: medRxiv

Article Title: Discovery of a radiation countermeasure therapeutic for intestinal injury enabled by human organ chips combined with AI

doi: 10.1101/2025.10.03.25337298

Figure Lengend Snippet: A) Photograph of an Organ Chip and a diagram showing how intestinal epithelial cells isolated from patient-derived ileal organoids and intestinal microvascular endothelial cells were cultured on opposite sides of a porous ECM-coated membrane that separates two parallel channels of a commercially available Organ Chip device to form epithelial-endothelium tissue interface. Culture medium is flowed through the upper epithelium-lined channel and lower endothelial channel to mimic movement of intestinal contents and vascular fluid flow, respectively. The epithelium.,,which is composed of absorptive cells and Goblet cells, forms villi-like structures and secretes a mucus layer into the lumen of the apical channel. The entire tissue-tissue interface can be stretched and relaxed rhythmically to mimic peristalsis-like motions by applying cyclic suction to hollow side chambers in the flexible device (shown with white arrows at left). B) DIC images of the villus epithelium when viewed from above in Ileum Chips after being exposed to radiation for 6 hours to 7 days. Radiation exposure results in a reduction in the number of villus-like structures (bar, 500 μm) . C) Immunofluorescence micrographs showing vertical cross-sections of Ileum Chips that demonstrate changes in the morphology of villus-like epithelial structures 7 days after different irradiation doses. Magenta, phalloidin-stained F-actin; white, DAPI-stained nuclei; Dashed yellow line, upper surface of chip membrane; dashed cyan line, upper luminal boundary of the villus (bar, 100 µm) . D) Higher magnification views of immunofluorescence micrographs from C showing regions of the ileal epithelium where it contacts the porous ECM-coated membrane on-chip (bar, 50 µm) E) Histological H&E stained vertical cross-sections confirming 16Gy radiation dose significantly reduces the height of the epithelial monolayer (bar, 50 µm). F) Immunofluorescence micrographs of vertical cross sections through the Ileum Chip showing 53bp1 staining (green) for double-stranded DNA breaks 7 days after exposures to different levels of radiation (bar, 50 µm). G) Graphs showing that intestinal barrier permeability measured with either 3 or 40 kDa Cascade Blue is significantly higher in Ileum Chips exposed to 16 Gy radiation for 7 days (n = 3 donors). Each symbol represents an individual health patient derived chip. H) Production of various pro-inflammatory cytokines and chemokines by Ileum Chips exposed to different radiation doses (n = 3 donors).

Article Snippet: Small intestine microvascular endothelial cells (HSIMECs, 10HU-065, iXCells Biotechnologies, CA) were expanded in endothelial basal medium-2 (Lonza CC-3156) supplemented with Primocin and bullet kit (Lonza CC-4176), which contains FBS, Hydrocortisone, hFGF-B, VEGF, R3-IGF-1, Ascorbic Acid, hEGF, Heparin.

Techniques: Isolation, Derivative Assay, Cell Culture, Membrane, Immunofluorescence, Irradiation, Staining, Permeability

Journal: Theranostics

Article Title: System-wide vitreous proteome dissection reveals impaired sheddase activity in diabetic retinopathy.

doi: 10.7150/thno.72947

Figure Lengend Snippet: Figure 6. ADAM10’s activator EGCG inhibits the activation of human retinal endothelial cells (HRECs) in vitro and vessel outgrowth in ex vivo models. (A and B) ADAM10 activity impairment in ocular disease models are shown. (A) ADAM10 activity in retinae from control mice and OIR mice. n = 3. (B) ADAM10 activity in choroid/RPE from control mice and CNV mice. n = 4. Data are presented as mean ± s.e.m. Statistical analysis was determined by unpaired, two-tailed Student’s t-test; * p < 0.05. (C) AlamarBlue assay demonstrated inhibition of HRECs viability by 50 µM EGCG following 24 h treatment (n = 3). (D) DAPI staining demonstrated inhibition of HRECs migration by 50 µM EGCG following 4 h treatment (n = 3). (E) EGCG inhibits choroidal vessel outgrowth in a dose-dependent manner. Representative images (left) and quantitative analysis (right) of microvessel formation from mouse choroidal explants demonstrating a significant inhibitory effect of EGCG (n = 3 independent experimental groups, n ≥ 6 explants per treatment group). (F) EGCG inhibits aortic vessel sprouts at the dosage of 50 µM. Representative images (left) and quantitative analysis (right) of macrovessel formation from mouse aortic explants demonstrating a significant inhibitory effect of EGCG (n = 3 independent experimental groups, n ≥ 6 explants per treatment group). Scale bar: 100 µm. All images shown are representative, and data are presented as mean ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test or unpaired, two-tailed Student’s t-test; * p < 0.05 and *** p < 0.001.

Article Snippet: Primary human retinal endothelial cells (HRECs) were purchased from Angioproteomie (Boston, MA, USA) and maintained in endothelial growth media 2 (EGM2), which contained EBM2 basal media and Endothelial Cell Growth Medium BulletKitsTM (Lonza, Basel, Switzerland) or endothelial basal media 2 (EBM2), which contained 0.2 % FBS according to the supplier’s instruction.

Techniques: Activation Assay, In Vitro, Ex Vivo, Activity Assay, Control, Two Tailed Test, Alamar Blue Assay, Inhibition, Staining, Migration, Comparison

Journal: Theranostics

Article Title: System-wide vitreous proteome dissection reveals impaired sheddase activity in diabetic retinopathy.

doi: 10.7150/thno.72947

Figure Lengend Snippet: Figure 7. AXL is ADAM10’s downstream substrate and ADAM10’s function is dependent on the activity of AXL. (A) Full-length AXL protein levels in HREC with 50 µM ADAM10 activator EGGC or 10 µM ADAM10 inhibitor GI 254023X is shown at both 4- and 24-h post treatment. GAPDH is used as loading control (B) Combination treatment of AXL inhibitor R428 and ADAM10 activator EGCG have no additive effects on HRECs viability. AlamarBlue assay demonstrated inhibition of HRECs viability by 50 µM EGCG, 1 µM R428 or combination treatment of 50 µM EGCG, 1 µM R428 following 24 h treatment (n = 3). (C) Combination treatment of AXL inhibitor R428 and ADAM10 activator EGCG have no additive effects on HRECs migration. DAPI staining demonstrated inhibition of HRECs migration by 50 µM EGCG, 5 µM R428 or combination treatment of 50 µM EGCG, 5 µM R428 following 4 h treatment (n = 3). (D) R428 reverses the promoting effect of EGCG inhibitor GI 254023X on HRECs viability. AlamarBlue assay demonstrated effects of HRECs viability by 10 µM GI 254023X, 1 µM R428 or combination treatment of 10 µM GI 254023X, 1 µM R428 following 24 h treatment (n = 3). (E) R428 reverses the promoting effect of GI 254023X on HRECs migration. DAPI staining demonstrated effects of HRECs migration by 10 µM GI 254023X, 5 µM R428 or combination treatment of 10 µM GI 254023X, 5 µM R428 following 4 h treatment (n = 3). Scale bar: 100 µm. All images shown are representative, and data are presented as means ± s.e.m. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparison test; ** p < 0.01 and *** p < 0.001.

Article Snippet: Primary human retinal endothelial cells (HRECs) were purchased from Angioproteomie (Boston, MA, USA) and maintained in endothelial growth media 2 (EGM2), which contained EBM2 basal media and Endothelial Cell Growth Medium BulletKitsTM (Lonza, Basel, Switzerland) or endothelial basal media 2 (EBM2), which contained 0.2 % FBS according to the supplier’s instruction.

Techniques: Activity Assay, Control, Alamar Blue Assay, Inhibition, Migration, Staining, Comparison

Journal: Cell Death & Disease

Article Title: Cinacalcet-mediated activation of the CaMKKβ-LKB1-AMPK pathway attenuates diabetic nephropathy in db/db mice by modulation of apoptosis and autophagy

doi: 10.1038/s41419-018-0324-4

Figure Lengend Snippet: a The changes of [Ca 2+ ]i in HGECs exposed to cinacalcet and high-glucose media. To determine whether the addition of cinacalcet might modulate [Ca 2+ ]i in HGECs, FURA-2AM-loaded HGECs were stimulated using different concentrations (15, 100 nM) of cinacalcet in low-glucose (LG; 5 mmol/l D-glucose) or high-glucose (HG; 30 mmol/l D-glucose) media. The area under curve (AUC) was estimated from the baseline of normalized data (at the point of injection) to a fluorescence level and between time points of injection (0 min) and 10 min. The peak of the curve was measured as highest value of the curve. The peak amplitude and AUC of [Ca 2+ ]i were significantly increased by cinacalcet in dose-dependent manners in both LG and HG media. In Fig. 1a, the arrow denotes the administration of cinacalcet (15 and 100 nM, respectively) ( n = 6 independent experiments in each experiments). * p < 0.05; ** p < 0.01 compared with LG and HG. b The changes of intracellular signaling in HGECs exposed to cinacalcet and high-glucose media. Representative immunofluorescent ( n = 6 independent experiments in each experiments) and western blot analyses ( n = 4 independent experiments in each experiments) of CaSR, CaMKKα/β, phospho-Ser 428 LKB1, and phospho-Thr 172 AMPK in the cultured HGECs in low-glucose (LG; 5 mmol/l D-glucose) or high-glucose (HG; 30 mmol/l D-glucose) conditions with or without cinacalcet treatment (15 nM) and the quantitative analyses of the results are shown. * P < 0.05; ** P < 0.01 and # P < 0.001 compared with other groups

Article Snippet: Human glomerular endothelial cells (HGECs) were purchased from Anigio-Proteomie (Boston, MA) and subcultured in endo-growth media (Angio-Proteomie, Boston, MA).

Techniques: Injection, Fluorescence, Western Blot, Cell Culture

Journal: Cell Death & Disease

Article Title: Cinacalcet-mediated activation of the CaMKKβ-LKB1-AMPK pathway attenuates diabetic nephropathy in db/db mice by modulation of apoptosis and autophagy

doi: 10.1038/s41419-018-0324-4

Figure Lengend Snippet: The effect of cinacalcet on intracellular signaling for AMPK-eNOS oxidative stress and apoptosis in the HGECs cultured in low-glucose (LG; 5 mmol/l D-glucose) or high-glucose (HG; 30 mmol/l D-glucose) conditions with or without cinacalcet treatment (1, 5, 15 nM) ( a–d ). Representative Western blot analyses and quantitative analyses of total AMPK, phosphor-Thr 172 AMPK, total eNOS, phospho-Ser 1177 eNOS ( a , * P < 0.05 and ** P < 0.01 compared with LG control), SOD1 and SOD2 ( b , * P < 0.05 compared with other groups), dihydroethidium expression (as an oxidative stress marker; c , * P < 0.05 and # P < 0.001 compared with other groups), Bcl-2, Bax, and TUNEL-positive HGECs ( e , * P < 0.05 and ** P < 0.01 compared with other groups), and β-actin levels in the cultured HGECs and their quantitative analyses of the results are shown ( n = 4 independent experiments in each experiments). d The effect of BAPTA-AM (25 μM) on cinacalcet-indueced in the HGECs cultured in low-glucose or high-glucose (HG; 30 mmol/l D-glucose) with or without cinacalcet treatment (15 nM). Representative Western blot analyses and quantitative analyses of CaMKKβ, phospho-LKB1, and total LKB1 ( n = 4 independent experiments in each experiments). * P < 0.05 and ** P < 0.01 compared with LG control. f The changes of intracellular signaling related to autophagy in HGECs exposed to cinacalcet and high-glucose media. Representative Western blot analyses and quantitative analyses of beclin-1, LC3-II/LC3-I ratio, and β-actin levels in the cultured HGECs and their quantitative analyses of the results are shown ( n = 4 independent experiments in each experiments). Representative immunofluorescent analyses of LC3 punctae in HGECs and the quantitative analyses of the results are shown ( n = 6 independent experiments in each experiments). ** P < 0.01 compared with other groups

Article Snippet: Human glomerular endothelial cells (HGECs) were purchased from Anigio-Proteomie (Boston, MA) and subcultured in endo-growth media (Angio-Proteomie, Boston, MA).

Techniques: Cell Culture, Western Blot, Control, Expressing, Marker, TUNEL Assay

Journal: Cell Death & Disease

Article Title: Cinacalcet-mediated activation of the CaMKKβ-LKB1-AMPK pathway attenuates diabetic nephropathy in db/db mice by modulation of apoptosis and autophagy

doi: 10.1038/s41419-018-0324-4

Figure Lengend Snippet: Immunoblot for CaMKKβ, LKB1, phospho-AMPK, SIRT1 and phospho-Ser 1177 eNOS in AMPKα1 siRNA, AMPKα2 siRNA, or SIRT1 siRNA knock-down HGECs in a high-glucose environment with cinacalcet treatment ( a and b ). The cultured HGECs were transfected with a final concentration of 50 nM CaMKKβ and LKB1, α1 and α2-AMPK, SIRT1 siRNAs for 24-h by transfection reagent and treated with cinacalcet (15 nM) in high-glucose media. Representative Western blot analyses of CaMKKβ and phospho-Ser 428 LKB1 ( a ), as well as phospho-Thr 172 AMPK, total AMPK, SIRT1, phospho-Ser 1177 eNOS ( b ) and β-actin levels and the quantitative analyses of the results are also shown ( a and b , respectively) ( n = 4 independent experiments in each experiments).* P < 0.05, ** P < 0.01 compared with control siRNA with HG

Article Snippet: Human glomerular endothelial cells (HGECs) were purchased from Anigio-Proteomie (Boston, MA) and subcultured in endo-growth media (Angio-Proteomie, Boston, MA).

Techniques: Western Blot, Knockdown, Cell Culture, Transfection, Concentration Assay, Control

Journal: Cell reports

Article Title: Cancer-associated fibroblasts maintain critical pancreatic cancer cell lipid homeostasis in the tumor microenvironment

doi: 10.1016/j.celrep.2024.114972

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Human pancreatic microvascular endothelial cells were cultured in Endothelial Cell Basal Medium (iXCells, MD-0010B) cultured for 48 h under 21% O 2 or 0.5% O 2 .

Techniques: Recombinant, Western Blot, Transfection, Reverse Transcription, Flow Cytometry, Bicinchoninic Acid Protein Assay, Plasmid Preparation, ROS Assay, Mass Spectrometry, shRNA, Control, Software, Microscopy, Real-time Polymerase Chain Reaction