Transcriptional roles of PARP1 in cancer.
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ABSTRACT:
SUBMITTER: Schiewer MJ
PROVIDER: S-EPMC4134958 | biostudies-literature | 2014 Aug
REPOSITORIES: biostudies-literature
Publications
Transcriptional roles of PARP1 in cancer.
Schiewer Matthew J MJ Knudsen Karen E KE
Molecular cancer research : MCR 20140610 8
Poly (ADP-ribose) polymerase-1 (PARP1) is an abundant, ubiquitously expressed NAD(+)-dependent nuclear enzyme that has prognostic value for a multitude of human cancers. PARP1 activity serves to poly (ADP-ribose)-ylate the vast majority of known client proteins and affects a number of cellular and biologic outcomes, by mediating the DNA damage response (DDR), base-excision repair (BER), and DNA strand break (DSB) pathways. PARP1 is also critically important for the maintenance of genomic integri ...[more]
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Project description:PurposePARP inhibitor (PARPi) therapy is approved for patients with metastatic castration-resistant prostate cancer (mCRPC) and homologous recombination repair (HRR) genomic aberrations. However, only a fraction of patients with BRCA1/2 mutations respond to PARPi therapy. In this pilot study, we assess PARP-1 expression in prostate cancer patients with and without HRR genomic alternations using a novel PARP-based imaging agent.ProceduresNine advanced prostate cancer patients were studied with PET/CT and [18F]FluorThanatrace (FTT), an analogue of the PARPi rucaparib. Images were analyzed using maximum standardized uptake values (SUVmax). PARP expression was assessed by immunohistochemistry (IHC) when feasible (n = 4).ResultsWe found great variability in FTT uptake (SUVmax range: 2.3-15.4). Patients with HRR mutations had a significantly higher SUVmax (p = 0.0379) than patients with non-HRR mutations although there was an overlap in FTT uptake between groups. Three patients without HRR and one with HRR mutations had similarly high PARP1 IHC expression.ConclusionsFTT-PET/CT may serve as an alternate biomarker for PARP1 expression and a potential method for PARPi treatment selection.
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Identification of DSSO Cross-Linked Peptides The m/z, charges from MS2 and corresponding m/z and intensities of fragment ion from MS3 spectra were extracted into ms2 file format using RawDistiller v. 1.0, in-house developed software with D(g, 6) settings to abstract parent ion profiles by Gaussian fitting and dynamic offline lock mass using six background polydimethylcyclosiloxane ions as internal calibrants. MS3 spectra were first searched using ProLuCID with a mass tolerance for parent ions and fragment ions set as 7 ppm and 800 ppm, respectively. Trypsin specificity was imposed on both ends of candidate peptides during the search against a protein database combining 4540 proteins including 426 common contaminants. The methionine oxidation (O; 15.9949 Da) and remnants of the crosslinker alkene (C3H2O; +54.0106 Da) and thiol (C3H2SO; +85.9826 Da) were set as variable modifications. A static modification with a mass of 57.0125Da was added to cysteine residues. A script module was added into NSAF7 to search MSn data and identify putative crosslinkers based on their unique MS fragmentation patterns with the false positive rate was not more than 1% as illustrated in. The Xcorr of the peptides for a specified cross-linker was summed as scores S and use log2(10*log2(S+1)+1) to calculate the cross-linker scores. The non-cross-linking peptide/spectrum matches were sorted and selected using DTASelect/CONTRAST (v. 1.9) with an in-house software, swallow to filter spectra, peptides, and proteins at FDRs <1%. NSAF7 was used to create the final reports on all detected cross-linkers, peptides and non-redundant proteins identified across the different runs.
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