Journal: Scientific Reports

Article Title: Pyrazine ring-based Na + /H + exchanger (NHE) inhibitors potently inhibit cancer cell growth in 3D culture, independent of NHE1

doi: 10.1038/s41598-020-62430-z

Figure Lengend Snippet: EIPA, but not cariporide, potently reduces viability of MCF-7 and MDA-MB-231 spheroids. MCF-7 and MDA-MB-231 spheroids were grown for 7–9 days. Spheroids were treated simultaneously with NHE1-inhibitors (cariporide (10 µM) or EIPA (10 µM)), breast cancer subtype specific anti-cancer therapy (tamoxifen (Tam, 2 µM) or chemotherapy (Cisplatin (18.75 nM), Doxorubicin (18.75 nM) and 5-FU (0.0625 nM)) or a combination thereof as indicated, on day 2 and 4 (MDA-MB-231) or 2, 4 and 7 (MCF-7). DMSO served as vehicle for the anti-cancer therapy. Light microscopic images of spheroids were acquired on day 2, 4, 7 (MDA-MB-231) and 9 (MCF-7). ( A , C ) Representative images of MCF-7 and MDA-MB-231 spheroids, respectively. 3–6 n. Scale bar: 100 µm. ( B , D ) Cell viability of MCF-7 (day 9) ( B ) and MDA-MB-231 (day 7) ( D ) spheroids, respectively. One-way ANOVA test with Tukey’s multiple comparisons post-test was used to determine statistically significant differences between treatment groups. * and # denotes significant differences between the treatment condition relative to DMSO and between two treatment conditions, respectively. For panel 1B, the p -value is 0.0116 and 0.0147 for control vs. tamoxifen + cariporide and tamoxifen vs. tamoxifen + EIPA, respectively, while for panel 1D the p -value is 0.0011 and 0.0143 for control vs. chemotherapy + cariporide and for EIPA vs. chemotherapy + EIPA, respectively. Error bars denote SEM. 3–6 n. ( E , G ) Representative traces demonstrating BCECF-fluorescence as a function of time during pH i recovery of MCF-7 (day 4) ( E ) and MDA-MB-231 (day 4) ( G ) spheroids after NH 4 Cl pre-pulse. Fluorescence is depicted as the 485/440 nm ratio of BCECF intensity. 2–4 n ( F , H ) Quantification of pH i recovery rate of MCF-7 ( p -value = 0.343 for control vs. both cariporide and EIPA) ( F ) and MDA-MB-231 ( p -value = 0.348 for control vs. cariporide, and 0.0461 for control vs. EIPA) ( H ) spheroids determined as the slope of the first 120 s after maximum acidification. Error bars denote SEM. 2–4 n. Statistical significance determined using a one-way ANOVA test with Tukey’s multiple comparisons test. 2–3 n.

Article Snippet: Cariporide was a gift from Sanofi-Aventis and 5-( N -ethyl- N -isopropyl) amiloride (EIPA) (#E3111) was from Thermo-Fisher. β-actin antibody (#A5441), tamoxifen (#T5648), cisplatin (#P4394), 5-fluorouracil (5-FU) (#F6627), 5-( N , N -Dimethyl) amiloride (DMA) (#A4562), 5-( N , N -Hexamethylene) amiloride (HMA) (#A9561) and amiloride (#A7410) were from Sigma-Aldrich, and doxorubicin (#120629) and eniporide (#HY-106150B) from Abcam and MedChemExpress, respectively.

Techniques: Control, Fluorescence

Journal: International Journal of Biological Sciences

Article Title: Integrative Single-Cell and Spatial Transcriptomics Analysis Reveals ECM-remodeling Cancer-associated Fibroblast-Derived POSTN as a Key Mediator in Pancreatic Ductal Adenocarcinoma Progression

doi: 10.7150/ijbs.108618

Figure Lengend Snippet: CAF-derived POSTN induces the translation to EMT-subtype via PI3K/AKT/β-catenin signaling in PDAC Cells. (A) Differential gene expression analysis of ductal cells based on fibroblast-derived POSTN levels. Tumor samples from the CRA001160 dataset were classified into high- and low- POSTN groups (75th percentile cutoff), and ductal cell gene expression profiles were compared to identify differences linked to CAF-derived POSTN. (B) GSEA of upregulated genes in ductal cells from POSTN-high samples identified enriched KEGG pathways. (C-F) GSEA of upregulated genes revealed enrichment in the EMT pathway, focal adhesion pathway, PI3K-AKT pathway, and WNT pathway. (G-H) Western blot analysis of β-catenin expression, and phosphorylation of FAK, AKT, and GSK-3β in BxPC-3 and PANC-1 cells after 24-hour treatment with rhPOSTN at varying concentrations. (I) Schematic illustration of how CAF-derived POSTN drives the EMT phenotype in PDAC cells via integrin αvβ5/FAK/PI3K/AKT/β-catenin signaling pathway.

Article Snippet: BxPC-3 cells were pretreated for 24 h with CAF-oePOSTN CM, CAF-NC CM, recombinant human Periostin (rhPOSTN, Novoprotein, CJ39), or integrin αvβ5 receptor inhibitor (MCE, HY-16141).

Techniques: Derivative Assay, Gene Expression, Western Blot, Expressing, Phospho-proteomics

Journal: International Journal of Biological Sciences

Article Title: Integrative Single-Cell and Spatial Transcriptomics Analysis Reveals ECM-remodeling Cancer-associated Fibroblast-Derived POSTN as a Key Mediator in Pancreatic Ductal Adenocarcinoma Progression

doi: 10.7150/ijbs.108618

Figure Lengend Snippet: Integrin αvβ5 inhibitors partially reverse POSTN-induced proliferation, colony formation, migration, and invasion of PDAC cells. (A) Co-localization of POSTN and intergrin β5 in PDAC tissues. Immunofluorescence staining showing POSTN (green), integrin β5 (red), and nuclei (blue) in PDAC patient resection specimens. Scale bars, 25 μm. (B-C) Effect of integrin αvβ5 inhibition on POSTN-induced proliferation in BxPC-3 and PANC-1 cells. Cells were treated with: (1) negative control, (2) 500 ng/mL rhPOSTN, (3) integrin αvβ5 inhibitor (HY-16141) at 1/5 IC 50 concentration, or (4) integrin αvβ5 inhibitor pretreated for 24 hours, followed by 500 ng/mL rhPOSTN. Cell proliferation was assessed using CCK-8 assays. (D) Colony formation assays in BxPC-3 and PANC-1 cells following the same treatments as in (B-C). (E-F) Wound healing assays in BxPC-3 and PANC-1 cells after treatments as described in (B-C), assessing migration capacity. (G-H) Transwell assays in BxPC-3 and PANC-1 cells to assess migation (G) and invasion (H) under the same treatment as in (B-C). (I) Western blot analysis of β-catenin expression, and phosphorylation of FAK, AKT, and GSK-3β in BxPC-3 and PANC-1 cells after 24-hour treatment as described in (B-C).

Article Snippet: BxPC-3 cells were pretreated for 24 h with CAF-oePOSTN CM, CAF-NC CM, recombinant human Periostin (rhPOSTN, Novoprotein, CJ39), or integrin αvβ5 receptor inhibitor (MCE, HY-16141).

Techniques: Migration, Immunofluorescence, Staining, Inhibition, Negative Control, Concentration Assay, CCK-8 Assay, Western Blot, Expressing, Phospho-proteomics

Journal: Advanced Science

Article Title: Macrophage TRIM21 Inhibition Ameliorates Murine Acute Pancreatitis via PHB2‐Mediated Mitochondrial Stabilization

doi: 10.1002/advs.202517877

Figure Lengend Snippet: Vilazodone‐mediated activation of TRIM21 exacerbated L‐arginine‐induced AP. (A‐G) Trim21 M‐WT mice pretreated with vehicle or vilazodone intraperitoneally received saline or L‐arginine for 72 h (Vehicle + Saline, Vehicle + L‐arginine, Vilazodone + Saline, Vilazodone + L‐arginine; n = 10 per group). (A) Serum amylase levels. (B) Representative gross morphology of the pancreas. (C) Pancreas‐to‐body weight ratio. (D) H&E staining (upper panel) and histopathological scores (lower panel) of the pancreatic tissue. (E) Representative images (upper panel) and quantification (lower panel) of IHC staining for IL‐1β and TNF‐α in pancreatic tissue. (F, G) IL‐1β and TNF‐α levels in serum samples and BALF. Data are expressed as mean ± SEM (A, C–G), and statistical analyses were conducted using ANOVA with Tukey HSD post hoc test (A, C), Kruskal‐Wallis test with Dunn's post hoc test (D), and Welch's ANOVA with Games‐Howell post hoc test (E–G).

Article Snippet: TRIM21 was activated using vilazodone (25 mg/kg; MedChemExpress) and downregulated using quisinostat (10 mg/kg; MedChemExpress).

Techniques: Activation Assay, Saline, Staining, Immunohistochemistry

Journal: International journal of molecular sciences

Article Title: Pharmacological Inhibition of NHE1 Protein Increases White Matter Resilience and Neurofunctional Recovery after Ischemic Stroke.

doi: 10.3390/ijms241713289

Figure Lengend Snippet: Figure 4. Efficacy of the post-stroke delivery of the novel NHE1 inhibitor Rimeporide. (A) Experimental protocol. (B,C) TTC staining and infarct volume and brain swelling analysis of the Veh- and RIM-treated mice at 3 d post-stroke. N = 3. Dotted line and arrow: infarct tissues. (D,E) Neurological scoring and rotarod accelerating test results for motor-sensory functions in Veh- or RIM-treated mice from 1 to 14 d post-stroke. N = 9 for Veh, N = 10 for RIM. (F–H) Open field test, Y-maze spontaneous alternation test (at 28-30 d post-stroke), and novel spatial recognition test results (at 37-40 d post-stroke) in the same cohort of mice in (D,E) Data are the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: HOE642 (Cariporide, Sigma-Aldrich, St. Louis, MO, USA) or Rimeporide (EMD-87580, MedChemExpress, Monmouth Junction, NJ, USA) was dissolved at 1 mg/mL in DMSO as stock solution and diluted to 0.025 mg/mL in PBS immediately before injection.

Techniques: Staining

Journal: International journal of molecular sciences

Article Title: Pharmacological Inhibition of NHE1 Protein Increases White Matter Resilience and Neurofunctional Recovery after Ischemic Stroke.

doi: 10.3390/ijms241713289

Figure Lengend Snippet: Figure 5. Post-stroke delivery of Rimeporide improved white matter myelination after stroke. (A) Representative images and quantitative analysis of MBP intensity and APC+ counts in the Veh- and RIM-treated brains at 3 d post-stroke. Arrows: APC+ cells. N = 3. (B) Representative images and quantitative analysis of NG2+Olig2+, Caspase3+Olig2+, and H3K9me3+Olig2+ cells in the CL and IL hemispheres of the CC at 3 d post-stroke. Arrows: double positive cells. N = 3. Data are the mean ± SEM. ** p < 0.01, *** p < 0.001.

Article Snippet: HOE642 (Cariporide, Sigma-Aldrich, St. Louis, MO, USA) or Rimeporide (EMD-87580, MedChemExpress, Monmouth Junction, NJ, USA) was dissolved at 1 mg/mL in DMSO as stock solution and diluted to 0.025 mg/mL in PBS immediately before injection.

Techniques:

Journal: International journal of molecular sciences

Article Title: Pharmacological Inhibition of NHE1 Protein Increases White Matter Resilience and Neurofunctional Recovery after Ischemic Stroke.

doi: 10.3390/ijms241713289

Figure Lengend Snippet: Figure 6. Post-stroke administration of HOE642 or Rimeporide reduced microglial inflammation without affecting astrogliosis (A). Representative images and quantitative analysis of GFAP+ and IBA1+ cells in the CL and IL cortex of Veh- and HOE-treated mice at 3 d post-stroke. (B) Representative gating strategy for CD11b+CD45+ microglia/macrophages. (C) Quantitative analysis of inflammatory profiling markers within parent CD11b+CD45+ microglia/macrophages in Veh- and HOE-treated mice at 3 d post-stroke. (D) Representative images and quantitative analysis of GFAP+ and IBA1+

Article Snippet: HOE642 (Cariporide, Sigma-Aldrich, St. Louis, MO, USA) or Rimeporide (EMD-87580, MedChemExpress, Monmouth Junction, NJ, USA) was dissolved at 1 mg/mL in DMSO as stock solution and diluted to 0.025 mg/mL in PBS immediately before injection.

Techniques:

Journal: The Journal of Clinical Investigation

Article Title: Aging-dependent microglial heterogeneity worsens outcomes in models of traumatic brain injury

doi: 10.1172/JCI196112

Figure Lengend Snippet: ( A ) High-throughput virtual screening was performed on 8,561 potential ELF1 inhibitor compounds approved by the FDA or already in clinical use. 200 compounds with the highest docking scores were screened after sequential SP and XP docking analysis. Finally, 10 compounds were screened out of all 200 compounds with the highest potential to cross the blood-brain barrier (BBB). ( B ) Docking scores, molecular weights, and clinical information for the 10 compounds selected. ( C ) Ratio of NLRP3 + microglia after Imeglimin treatment (HMC3). n = 3/group. ( D ) Chemical structure of Imeglimin and in silico docking to the active pocket of the human ELF1 protein. ( E and F ) ECAR and OCR of Imeglimin-treated microglia cell line (HMC3) and vector controls. n = 3/group. ( G ) Flow chart of Imeglimin in vivo treatment of mice with TBI. ( H ) Survival curves of Imeglimin-treated mice versus vector control mice after 45 days of TBI (left, young group; right, aged group; start: n Sham = 12, n Vector+Sham = 20, and n Ime+TBI = 20). ( I and J ) mNSS ( I ) and rotarod test ( J ) between Imeglimin-treated and vector control mice after 45 days of TBI (young group, n Sham = 12, n Vector+TBI = 17, and n Ime+TBI = 18; aged group, n Sham = 12, n Vector+TBI = 13, and n Ime+TBI = 19). ( K ) Concentrations of IL-1β and IL-6 in the cerebrospinal fluid of Imeglimin-treated and vector control mice after 45 days of TBI ( n Young-vector = 17, n Aged-vector = 13, n Young-Ime = 18, and n Aged+Ime = 19). Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01, **** P < 0.0001. Statistical analyses were performed using unpaired 2-tailed Student’s t test ( C ), 2-way ANOVA followed by Tukey’s post hoc test ( I – K ), and Kaplan-Meier survival analysis ( H ). The schematic was generated using BioRender.

Article Snippet: Imeglimin (MedChemExpress, HY-14771A) was dissolved in 10% DMSO and 90% corn oil.

Techniques: High Throughput Screening Assay, In Silico, Plasmid Preparation, In Vivo, Control, Generated

Journal: Pharmaceutics

Article Title: Development of Imeglimin Electrospun Nanofibers as a Potential Buccal Antidiabetic Therapeutic Approach

doi: 10.3390/pharmaceutics15041208

Figure Lengend Snippet: SEM images that show ( a ) blank nanofibers and ( b ) imeglimin-loaded nanofibers as smooth, non-porous, and non-beaded, with average fibers diameters of 450 ± 82 nm and 361 ± 54 nm, respectively (n = 70). The diameter distribution of ( c ) blank nanofibers and ( d ) imeglimin-loaded nanofibers show the average diameters of the fibrous systems.

Article Snippet: Imeglimin hydrochloride (CAS No. 775351-61-6) was bought from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Development of Imeglimin Electrospun Nanofibers as a Potential Buccal Antidiabetic Therapeutic Approach

doi: 10.3390/pharmaceutics15041208

Figure Lengend Snippet: XRD patterns of imeglimin, PM, blank, and imeglimin-loaded nanofibers showing that imeglimin was in the crystalline form (presence of characteristic reflections at 2θ: 14.71°, 16.82°, 24.36°, 24.87°, 25.42°, 31.64°, and 34.03°, which are reflected by the green arrows) as pure and in the PM. The blank and drug-loaded nanofibers were amorphous (broad halo pattern). The absence of the imeglimin distinct peaks in the drug-loaded nanofibers suggests the molecular dispersion of the drug due to the electrospinning process. DL: drug-loaded, PM: physical mixture.

Article Snippet: Imeglimin hydrochloride (CAS No. 775351-61-6) was bought from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques: Dispersion

Journal: Pharmaceutics

Article Title: Development of Imeglimin Electrospun Nanofibers as a Potential Buccal Antidiabetic Therapeutic Approach

doi: 10.3390/pharmaceutics15041208

Figure Lengend Snippet: The disintegration of ( a ) blank nanofibers and ( b ) imeglimin-loaded nanofibers demonstrates that both nanofibrous systems dissolved ultra-rapidly (≤2 ± 1 s, n = 3).

Article Snippet: Imeglimin hydrochloride (CAS No. 775351-61-6) was bought from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Development of Imeglimin Electrospun Nanofibers as a Potential Buccal Antidiabetic Therapeutic Approach

doi: 10.3390/pharmaceutics15041208

Figure Lengend Snippet: The developed HPLC calibration curves of imeglimin show its excellent linearity (R 2 = 0.9992).

Article Snippet: Imeglimin hydrochloride (CAS No. 775351-61-6) was bought from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques:

Journal: Pharmaceutics

Article Title: Development of Imeglimin Electrospun Nanofibers as a Potential Buccal Antidiabetic Therapeutic Approach

doi: 10.3390/pharmaceutics15041208

Figure Lengend Snippet: The release profile of the imeglimin-loaded nanofibers. The drug release profile of the thermo-shaking incubator method ( a ) showed a burst release at 1 min and a full drug release after 30 min. The drug release profile of the Franz diffusion method ( b ) showed a sustained release of imeglimin from approximately up to 6 h and a complete drug release after 24 h. The results are presented as the mean ± SD (n = 3).

Article Snippet: Imeglimin hydrochloride (CAS No. 775351-61-6) was bought from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques: Diffusion-based Assay

Journal: Pharmaceutics

Article Title: Development of Imeglimin Electrospun Nanofibers as a Potential Buccal Antidiabetic Therapeutic Approach

doi: 10.3390/pharmaceutics15041208

Figure Lengend Snippet: Cell viability of different doses of imeglimin upon 24 h HFF-1 cells exposure. The data demonstrated that imeglimin is safe at ≤250 µg/mL. Results are presented as mean ± SD (n = 3).

Article Snippet: Imeglimin hydrochloride (CAS No. 775351-61-6) was bought from Med Chem Express (Monmouth Junction, NJ, USA).

Techniques: