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