Journal: International Journal of Biological Sciences

Article Title: ARID1A deficiency activates OSM-STAT3 axis in endometrial cancer, creating vulnerability to JAK/STAT3 inhibition

doi: 10.7150/ijbs.129142

Figure Lengend Snippet: Identification of synthetic lethality of ARID1A and JAK/STAT3 in endometrial cancer through drug library screening. A. Schematic illustration of the synthetic lethality screenings with the Epigenetics Drug Library and the Kinase Inhibitor Library. B. Immunoblot analysis showing ARID1A knockout in A1 and B7 single clone. C. Drugs with selectivity index (SI)≥2 were selected as synthetic lethality candidates. SI=IC50 ARID1A+/+ /IC50 ARID1A-/- . D. Synthetic lethality in HEC1B-ARID1A -/- cell treated with stattic (D, F) and gandotinib (E, G) for 72h. The cell images were taken with IncuCyte ZOOM. Scale bar, 300 μm. H. Immunoblot analysis validated the ARID1A overexpression in HEC1B-ARID1A -/- cell. I. Overexpressed ARID1A rescued the synthetic lethality mediated by stattic. Data are mean±sd. * P <0.05, ** P <0.01. J-K. HEC1B ARID1A isogenic cell pair were transfected with STAT3-siRNA for 72 h. J. The quantitative analysis of synthetic lethality induced by STAT3-siRNA. Data are mean±sd. *** P <0.001. K. The growth curve of HEC1B ARID1A isogenic cell pair treated with STAT3-siRNA. **** P <0.0001. Two-way ANOVA test. L- N. IC50 test treated with stattic and gandotinib in endometrial cancer cell line. Data are mean±sd. * P <0.05.

Article Snippet: Epigenetics Drug Library and the Kinase Inhibitor Library screening were purchased from Selleck Chemicals.

Techniques: Drug discovery, Western Blot, Knock-Out, Over Expression, Transfection

Journal: Frontiers in Neuroscience

Article Title: Selected Histone Deacetylase Inhibitors Reverse the Frataxin Transcriptional Defect in a Novel Friedreich’s Ataxia Induced Pluripotent Stem Cell-Derived Neuronal Reporter System

doi: 10.3389/fnins.2022.836476

Figure Lengend Snippet: Screening of epigenetic compound library to identify novel inducers of FXN expression. (A) An overview of the screening process. All compounds of the APExBIO DiscoveryProbe™ Epigenetics Compound Library were tested at 10 μM concentration. Luminescence signal was plotted independently per each plate. The green line corresponds to plate average signal; dashed lines indicate average signal plus 1, 2, and 3 standard deviations of the mean. All data from two rounds of screening are included as . (B) Secondary validation of the efficacy of 16 compounds selected by initial screens. Luminescence detection was performed after 24 and 48 h of treatment. DMSO- and RG109-treated FXN-NLuc NPCs were used as controls. Five compounds were selected for further studies: CI994, Mocetinostat, Entinostat, UF 010, and Cerdulatinib. Results are mean ± SD from three technical replicates; * indicates p < 0.05 by one-way ANOVA. (C) Dose–response analyses for the selected compounds. Luminescence analyses were performed after 48 h of treatment with compounds at 0.5, 1, 2, 5, and 10 μM. EC50 is indicated for each compound. (D) Determination of cytotoxicity in FXN-NLuc NPCs using an LDH assay. Cytotoxicity is calculated relative to the spontaneous LDH release detected in DMSO-treated cells. Cells were treated with 10 μM of each compound for 48 h. Pirarubicin served as a positive control for cytotoxicity. Results are mean ± SD from three independent experiments; *** indicates p < 0.001 by one-way ANOVA.

Article Snippet: Next, as a proof of concept, we performed a luciferase-based screening of the Discovery Probe Epigenetic Compound Library (APExBIO) that includes 281 compounds predominantly targeting various epigenetic and chromatin pathways ( ).

Techniques: Drug discovery, Expressing, Concentration Assay, Biomarker Discovery, Lactate Dehydrogenase Assay, Positive Control

Journal: Frontiers in Oncology

Article Title: PBRM1 Deficiency Sensitizes Renal Cancer Cells to DNMT Inhibitor 5-Fluoro-2’-Deoxycytidine

doi: 10.3389/fonc.2022.870229

Figure Lengend Snippet: Epigenetic compound library screen identifies DNMT inhibitors as synthetic lethal drugs in PBRM1-deficient renal cancer cells. (A) Western Blot analysis showing loss of PBRM1 expression in the three PBRM1−/− clones. (B) The genomic Sanger sequencing of PBRM1 locus in 786-O PBRM1+/+ and PBRM1-/-(#1) cells. PBRM1−/− clone#1 lost 25 nucleotides in exon 3. (C) A compound inhibition rate plot of the first round screen data are shown. (D) A log10-IC50 plot of the second round screen data are shown. The IC50 values of the compounds against 786-O PBRM1+/+ and PBRM1-/- cells was plotted. Compounds with selectivity index (SI) > 2 for PBRM1−/− cells were chosen as synthetic lethality candidates. (E–H) Cell viability assay was done to certify the synthetic lethality effect by Fdcyd in 786-O isogenic pairs (E) and CAKI-1 isogenic pairs (F) . The other two DNMTis 5-Azacytidine (G) and Decitabine (H) were also used to test the IC50 in 786-O isogenic pairs. Error bars represent s.d. (n = 9) from three independent experiments. ANOVA P value of <0.001 for Fdcyd, 5-Azacytidine and Decitabine.

Article Snippet: Epigenetics Compound Library (L1200, 773 epigenetics compounds) was purchased from Topscience.

Techniques: Drug discovery, Western Blot, Expressing, Clone Assay, Sequencing, Inhibition, Viability Assay