Journal: Journal of Medicinal Chemistry

Article Title: RTx-303, an Orally Bioavailable Polθ Polymerase Inhibitor That Potentiates PARP Inhibitors in BRCA Mutant Tumors

doi: 10.1021/acs.jmedchem.5c00551

Figure Lengend Snippet: Cellular and in vitro activity of RTx-284. A. Bar plot showing the IC50 of RTx-284 in the indicated HR-proficient (gray) and HRD (red) cell lines. Data represent mean of at least 3 independent experiments performed in triplicate. B-D. Synergy plots created by Combenefit showing synergistic activity between RTx-284 and indicated PARPi in the indicated HRD cell lines. E. Bar plot showing that 10 μM RTx-284 suppresses the MMEJ GFP reporter. Data represent mean of 3 individual experiments performed in triplicate ±SEM. F. Dot plot showing increase of γH2AX following RTx-284:olaparib treatment in DLD1 BRCA2–/– cells. G. Bar plot showing that RTx-284 promotes an increase in caspase positive DLD1 BRCA2–/– cells. Data represent mean of 3 independent experiments performed in triplicate, ±SEM.

Article Snippet: HCT 116 BRCA2 −/– and HCT 116 Parental were obtained from Cancertools, London, UK.

Techniques: In Vitro, Activity Assay

Journal: Journal of Medicinal Chemistry

Article Title: RTx-303, an Orally Bioavailable Polθ Polymerase Inhibitor That Potentiates PARP Inhibitors in BRCA Mutant Tumors

doi: 10.1021/acs.jmedchem.5c00551

Figure Lengend Snippet: RTx-303 potentiates PARPi in BRCA mutant cells and xenografts. A. Scatter plot showing relative viability of the indicated ID8 cells treated with the indicated concentrations of olaparib with and without 5 μM RTx-303. Data represent mean of 3 independent experiments performed in triplicate, ±SEM. B. Scatter plot showing BRCA2 mutant BR-05–0566 PDX volumes following treatment with vehicle, 45 mg/kg olaparib (PO,QD), 60 mg/kg RTx-303 (PO,BID), and 45 mg/kg olaparib (PO,QD) with 60 mg/kg RTx-303 (PO,BID) (left). Scatter plot showing % body weight change (right). Data represent mean, n = 7 ±SEM. C. Scatter plot showing BRCA2 mutant BR-05–0568 PDX volumes following treatment with vehicle, 0.12 mg/kg talazoparib (PO,QD) with and without 60 mg/kg RTx-303 (PO,BID) (left). Scatter plot showing % body weight change. (right). Data represent mean, n = 6 ±SEM. D. Scatter plot showing MDA-MB-436 tumor volumes following treatment with vehicle, 45 mg/kg olaparib (PO,QD) with and without 60 mg/kg RTx-303 (PO,BID) (left). Scatter plot showing % body weight change (right). Data represent mean, n = 8 ±SEM. * p < 0.05, *** p < 0.001.

Article Snippet: HCT 116 BRCA2 −/– and HCT 116 Parental were obtained from Cancertools, London, UK.

Techniques: Mutagenesis

Journal: iScience

Article Title: Histone H2A ubiquitination resulting from Brap loss of function connects multiple aging hallmarks and accelerates neurodegeneration.

doi: 10.1016/j.isci.2022.104519

Figure Lengend Snippet: Figure 6. Histone H2A ubiquitination accompanied by BRCA1 activation is the hallmark phenotype of BRAP LOF (A) Immunoblotting of histone extracts from NPCs as well as from embryonic, neonatal, adult cerebral cortical tissues, and quantification (Mean G SD) of increases in histone H2Aub (total H2Aub and H2AubK119, respectively) resulted from Brap LOF. n = 3–6 biological replicates. p-values calculated by Student’s t test are indicated. (B) Immunoblotting of Brca1 in various cells and tissues, showing that Brap LOF results in increased Brca1 abundance. (C) Immunoblotting of nuclear vs cytoplasmic fractions of MEFs at P1, showing increased nuclear localization of Brca1 in Brap/ cells. (D) Brca1 (red) and NeuN (green) double immunohistology images of cerebral cortical sections of BrapcKONPC and control mice at four months of age. Representative images are shown. Note the increased intensity and density of Brca1 puncta in the nuclei of BrapcKONPC cortical neurons (NeuN+). (E and F) Immunoblotting analyses of histone extracts from cerebral cortical tissues of three-month-old mice, showing increased ubiquitination of H2A variants targeted by Brca1 (E) along with total histone H2A ubiquitination (F). (G) Double immunohistology staining of cortical sections of 4-month old WT or BrapcKONPC mice with antibodies against Gfap (green) and histone H3 (red), showing reduced nuclear histones in cells surrounded by reactive astrocytes (circles) in BrapcKONPC cortical tissues. Representative images are shown. Nuclear DNA was stained with Hoechst 33342. Bars: 50 um or as indicated. See also Figure S3.

Article Snippet: Phospho-p53 (Ser15) Abcam Cat# ab1431; RRID:AB_301090 Phospho-p53 (Ser15) (D4S1H) Cell Signaling Technology Cat# 12571; RRID:AB_2714036 Phospho-p53 (Ser15) (16G8) Cell Signaling Technology Cat# 9286; RRID:AB_331741 Phospho-ATM(Ser1981) Novus Biologicals Cat# AF1655 Phospho-ATR (Ser 428) Cell Signaling Technology Cat# 2853; RRID:AB_2290281 53 BP1 Novus Biologicals Cat# NB100-304 Brca1 Santa Cruz Biotechnology Cat# sc-642; RRID:AB_630944 Brca1 Novus Biologicals Cat# MAB22101 Brca1 (clone 8F7) Novus Biologicals Cat# NBP1-41186 LC3B (G-9) Santa Cruz Biotechnology Cat# sc-376404; RRID:AB_11150489 LC3B Cell Signaling Technology Cat# 2775; RRID:AB_915950 LC3B Novus Biologicals Cat# NB100-2220 p62SQSTM1 Novus Biologicals Cat# MAB8028 p62SQSTM1 GeneTex Cat# GTX100685; RRID:AB_2038029

Techniques: Ubiquitin Proteomics, Activation Assay, Western Blot, Control, Staining

Journal: Cancers

Article Title: Single-Nucleotide Polymorphisms of BRCA1 and BRCA2 and Risk of Papillary Thyroid Carcinoma

doi: 10.3390/cancers17091456

Figure Lengend Snippet: ( A ) Schematic of the BRCA1 DNA repair-associated ( BRCA1 ) gene on chromosome 17q21.31. ( B ) Physical map of BRCA2 DNA repair-associated ( BRCA2 ) gene on 13q13.1.

Article Snippet: , rs1799966 , C_2615208_20.

Techniques:

Journal: Cancers

Article Title: Single-Nucleotide Polymorphisms of BRCA1 and BRCA2 and Risk of Papillary Thyroid Carcinoma

doi: 10.3390/cancers17091456

Figure Lengend Snippet: ( A ) Linkage disequilibrium (LD) of single-nucleotide polymorphisms of BRCA1. ( B ) LD of single-nucleotide polymorphisms of BRCA2. The LD was based on the genotypes of the Korean population. In the Haploview program, the D value is expressed as a value between 0 and 100. The value 98 means 0.98. A red diamond without a number represents 1.

Article Snippet: , rs1799966 , C_2615208_20.

Techniques:

Journal: Cancers

Article Title: Single-Nucleotide Polymorphisms of BRCA1 and BRCA2 and Risk of Papillary Thyroid Carcinoma

doi: 10.3390/cancers17091456

Figure Lengend Snippet: The association results of BRCA1 variants and the risk of PTC.

Article Snippet: , rs1799966 , C_2615208_20.

Techniques: