Journal: Molecular Oncology

Article Title: PARP inhibitors elicit distinct transcriptional programs in homologous recombination competent castration‐resistant prostate cancer

doi: 10.1002/1878-0261.70098

Figure Lengend Snippet: Clinical PARPi elicit distinct antitumor activity in HR‐competent, PCa model systems. (A) TNMplot.com expression of PARP1 between normal prostate ( n = 106) and tumoral prostate ( n = 283) tissue ( P = 2.3e‐04). (B, C) PARP1 (B) and PAR (C) scores for Gleason 7–8 PCa TMA samples in EA ( n = 10) vs. AA ( n = 38) patients. AA, African American; EA, European American. (D, E) PARPi (Veliparib, Rucaparib, Olaparib, Niraparib, or Talazoparib) IC50's in C4‐2 (AR + , CRPC) (D) and DU145 (AR − , CRPC). CRPC, Castrate Resistant Prostate Cancer. Cells were treated vehicle control or a PARPi for 72 h. dsDNA Quantiflor One assay was performed to generate IC50 curves. Crystal violet validation of PARPi IC50 underneath each graph and in Fig. . Error bars represent standard deviation (SD) (D, E) from three, independent experiments. (F, G) Immunoblots of PARP1 and PAR levels after treatment with PARPi IC50 values found in C4‐2 (F) and DU145 (G). Normalized to b‐Actin. N, Niraparib; O, Olaparib; R, Rucaparib; T, Talazoparib; V, Veliparib. Representative of three, independent replicates in C4‐2 ( n = 3) and DU145 ( n = 3). (H) C4‐2 treated with regular growth conditions (left) or steroid depletion conditions (right) ± PARPi. Time points of 0, 48 and 96 h were analyzed for cell growth with the Incucyte. Error bars represent the standard error of the mean (SEM) between four, independent experiments. Each experiment had three biological replicates per experiment.

Article Snippet: Cells were treated as indicated with the PARP inhibitors (PARPi) Olaparib (Selleck S1060; Houston, TX, USA), Talazoparib (Selleck S7048), Rucaparib (Selleck S4948), Niraparib (Selleck S2741), or Veliparib (Selleck S1004).

Techniques: Activity Assay, Expressing, Control, Biomarker Discovery, Standard Deviation, Western Blot

Journal: Molecular Oncology

Article Title: PARP inhibitors elicit distinct transcriptional programs in homologous recombination competent castration‐resistant prostate cancer

doi: 10.1002/1878-0261.70098

Figure Lengend Snippet: Clinical PARPi elicit both overlapping and distinct alterations in gene expression. (A) Experimental design for treatment with PARPi. C4‐2 cells were steroid depleted for 72 h before introducing either vehicle control (DMSO) or a PARPi (Veliparib, Rucaparib, Olaparib, Niraparib, Talazoparib) to the cells. PARPi, PARP1 inhibitor. RNA sequencing was performed on each condition (with Novogene). Experiments are the result of four independent experiments. (B) Volcano plots of differentially expressed genes in each PARPi treatment condition relative to vehicle control. (C) Lotus plot of the number of DEGs in each PARPi treatment condition relative to vehicle control.

Article Snippet: Cells were treated as indicated with the PARP inhibitors (PARPi) Olaparib (Selleck S1060; Houston, TX, USA), Talazoparib (Selleck S7048), Rucaparib (Selleck S4948), Niraparib (Selleck S2741), or Veliparib (Selleck S1004).

Techniques: Gene Expression, Control, RNA Sequencing

Journal: Molecular Oncology

Article Title: PARP inhibitors elicit distinct transcriptional programs in homologous recombination competent castration‐resistant prostate cancer

doi: 10.1002/1878-0261.70098

Figure Lengend Snippet: p53‐related pathways are enriched in response to Olaparib and Niraparib treatment in a p53 WT model system. (A) Venn Diagram of the number of differentially expressed genes after Olaparib or Niraparib treatment. (B) Dot plot of TP53‐related Reactome pathways enriched in Olaparib‐ or Niraparib‐treated C4‐2. (C) Heatmap of TP53‐regulated genes in Olaparib‐ and Niraparib‐treated C4‐2 compared to vehicle control. (D, E) Relative expression of CDKN1A and DDB2 targets and immunoblot of p53 protein levels at 6 and 24 h posttreatment in C4‐2 (p53 WT) and DU145 (p53 mutant) cell lines. Immunoblot is representative of three independent replicates ( n = 3) in C4‐2 and DU145. Error bars on qPCR graph represent standard deviation (SD) between (D, E) three independent experiments in C4‐2 ( n = 3) and DU145 ( n = 3). N, Niraparib; O, Olaparib; Veh, Vehicle. Student's T ‐test was used to determine statistical significance from vehicle. * P < 0.05, ** P < 0.01, ns, not significant. For C4‐2, Olaparib CDKN1A P ‐value: 0.0032, Niraparib CDKN1A P ‐value: 0.0015. For C4‐2, Olaparib DDB2 P ‐value: 0.0014, Niraparib DDB2 P ‐value: 0.0164. For DU145, Olaparib CDKN1A P ‐value: 0.5558, Niraparib CDKN1A P ‐value: 0.0020. For DU145, Olaparib DDB2 P ‐value: 0.8777, Niraparib DDB2 P ‐value: 0.7795.

Article Snippet: Cells were treated as indicated with the PARP inhibitors (PARPi) Olaparib (Selleck S1060; Houston, TX, USA), Talazoparib (Selleck S7048), Rucaparib (Selleck S4948), Niraparib (Selleck S2741), or Veliparib (Selleck S1004).

Techniques: Control, Expressing, Western Blot, Mutagenesis, Standard Deviation

Journal: Molecular Oncology

Article Title: PARP inhibitors elicit distinct transcriptional programs in homologous recombination competent castration‐resistant prostate cancer

doi: 10.1002/1878-0261.70098

Figure Lengend Snippet: Genetic and therapeutic manipulation of p53 alters PARPi response. (A) Relative fold change 92 h post PARP inhibitor treatment in regular growth conditions with non‐targeting si‐RNA or si‐TP53. P ‐values are in comparison to si‐control vehicle control or si‐p53 vehicle control. P ‐values for relevant comparisons are reported from Student's T ‐test. Error bars represent SEM between four independent experiments. ** P < 0.001, **** P < 0.00001, ns, not significant. Si‐con + Olaparib P ‐value: 0.0012, si‐con + Niraparib P ‐value: < 0.0001, si‐p53 + Olaparib P ‐value: 0.3492, si‐p53 + Niraparib P ‐value: 0.1109. (B) Immunoblot validation of C4‐2 si‐TP53 knockdown representative of three independent replicates ( n = 3). N, Niraparib; O, Olaparib; Veh, Vehicle. (C) RT‐qPCR of CDKN1A expression in non‐targeting si‐RNA versus si‐TP53 conditions after treatment with vehicle control or PARP inhibitor. Si‐control + DMSO and si‐TP53 + DMSO were set to 1. DMSO, dimethyl sulfoxide (vehicle); NIRAP, Niraparib; OLAP, Olaparib. Student's T ‐test was used to generate P ‐values. Error bars represent SD (standard deviation) between three independent experiments. (D) DU145 IC50 value for PEITC. PEITC, Phenethyl isothiocyanate. Quantified with dsDNA Quantiflor 1. Error bars represent standard deviation between three independent experiments. (E) Synergy chart generated with CompSci after treatment with Olaparib and/or PEITC. Synergy experiments were based on results from nine independent experiments. (F) CDKN1A and DDB2 expression across normal, tumoral, and metastatic prostate tissue from TNMplot.com . Normal ( n = 106), Tumor ( n = 283), Metastatic ( n = 6).

Article Snippet: Cells were treated as indicated with the PARP inhibitors (PARPi) Olaparib (Selleck S1060; Houston, TX, USA), Talazoparib (Selleck S7048), Rucaparib (Selleck S4948), Niraparib (Selleck S2741), or Veliparib (Selleck S1004).

Techniques: Comparison, Control, Western Blot, Biomarker Discovery, Knockdown, Quantitative RT-PCR, Expressing, Standard Deviation, Generated

Journal: Cell Death & Disease

Article Title: Mitochondria-targeted oligomeric α-synuclein induces TOM40 degradation and mitochondrial dysfunction in Parkinson’s disease and parkinsonism-dementia of Guam

doi: 10.1038/s41419-024-07258-5

Figure Lengend Snippet: A Representative immunoblot of whole-cell extract from control NPSCs exposed to 10 µM 6OHDa for 24 h and subsequently treated with 10 µM Veliparib (PARPi) for an additional 24 h. B , C Densitometry analysis revealed a significant recovery in TOM40 protein level and reduced α-Syn protein level (Ab: α-Syn 204, Biolegend) following PARPi treatment (Ln 3). D Impact of PARPi and TOM40 overexpression on cell viability in SH-SY5Y cells under 6OHDA-induced toxicity. E Effect of PARPi and TOM40 overexpression on cell viability in WT α-Syn cells induced with dox for 48 h. F Schematic diagram illustrating mechanisms of TOM40 loss induced by α-Syn pathogenesis: ( a ) Normal protein import into mitochondria under physiological conditions. ( b ) In the presence of genetic mutations or neurotoxicants, α-Syn accumulates and forms oligomers (1), triggering TOM40 degradation via the UPS pathway (2), leading resulting in mitochondrial dysfunction (3). Inhibition of PARP1 serves to restore TOM40 levels, representing a promising strategy to counteract mitochondrial dysfunction and prevent cell death triggered by ROS toxicity and pathological α-Syn accumulation (4). Data ( B – E ) are presented as mean ± s.e.m. from three independent experiments and were analyzed by one-way ANOVA.

Article Snippet: NPSCs were subjected to a 24-h treatment with 10 µM 6OHDA, followed by a media change to either fresh media alone or media containing 10 µM Veliparib, a PARP inhibitor (PARPi) (MEDChemExpress, HY-10129, USA).

Techniques: Western Blot, Control, Over Expression, Inhibition