Enterprise | Interest None Epigenetics and prostate cancer - - PowerPoint PPT Presentation
Enterprise | Interest None Epigenetics and prostate cancer Defining the timing of DNA methyltransferases deregulation during prostate cancer progression Vassiliki Tzelepi 1 , Souzana Logotheti 1 , Efthymia Papakonstantinou 1 , Ioannis Maroulis
Enterprise | Interest None
Vassiliki Tzelepi1, Souzana Logotheti1, Efthymia Papakonstantinou1, Ioannis Maroulis2, Maria Melachrinou1 Vassiliki Zolota1
Departments of 1Pathology and 2Surgery, Medical School, University of Patras, Patras, Greece
changes in DNA sequence
DNA methylation Post-transcriptional histone modifications microRNAs expression
Chen et al. ONCOLOGY REPORTS 31: 523-532, 2014
Gravina et al. Molecular Cancer 9:305, 2010
DNA methyltransferases (DNMTs) DNMT1 DNMT2 DNMT3Α DNMT3Β DNMT3L
Α covalent chemical modification resulting in addition of a methyl group at the carbon 5 position of the cytosine ring in CpG dinucleotides
DNMT1: maintenance of basal methylation, passage of methylation to the daughter cells, methylation during cell proliferation DNMT2: tRNA methylation, role in DNA methylation? DNMT3Α DNMT3Β DNMT3L
De novo methylation
Tosoian et al. Nature Reviews Urology 13:205–215, 2016
Watson et al Nat Rev Cancer. 15:701–711, 2015
Synaptophysin Androgen Receptor
Small cell carcinoma Low PSA (for the extent of the disease) Visceral metastases Osteolytic bone metastases NO AR expression, no response to hormonal therapy Initially responds to chemotherapy, but resistance emerges VERY AGGRESSIVE
PCa cell lines and tissues
PCa cell lines
PCa in murine models
inhibition reduces tumor growth in PCa cell lines
Beltran H et al. Nat Med. 2016 22(3):298305 Kleb et al Epigenetics, 2016
CpG methylation CpG methylation
“Methylation status is complementary to morphology to discriminate AR dependent and independent CRPC”
compared to BPH (20 samples) and in high Gleason Score compared to low Gleason Score localized tumors (40 samples, 56 samples)
metastatic PCA (90 samples)
samples)
DNA methyltransferases are implicated in prostate cancer progression and the emergence
aggressive phenotypes
Elucidate the role of DNMTs in prostate cancer progression in an effort to identify prognostic markers and potential therapeutic targets
Low grade prostate cancer (GS: 6, 7(3+4), Prognostic Grade Group 1,2) High grade prostate cancer (GS: ≥7 (4+3), Prognostic Grade Group 3,4,5) Prostate cancer treated with androgen ablation Castrate Resistant Prostate Cancer Neuroendocrine prostate carcinoma received prostatectomy at MD Anderson cancer center
were selected and tissue microarrays were constructed.
– 0,6mm tissue cores (non-neoplastic peripheral zone and neoplasm) – 3069 cores (2637 neoplastic and 432 non- neoplastic) from 350 blocks – 2-17 cores per patient (mean: 6, SD: 3, median: 5) – 6 TMA blocks
DNMT1 DNMT2 DNMT3b
Company/ clone Santa Cruz, Heidelberg, Germany, H-12
(Mouse monoclonal)
Santa Cruz, Heidelberg, Germany, H-271
(Rabbit polyclonal)
Abcam, Cambridge, UK 52A1018
(Mouse monoclonal)
Antigen retreival EDTA, pH:8 Citrate, pH:6 EDTA, pH:8 Dilution 1:50 1:150 1:250 Incubation time 2 hours
30 minutes Detection System Envision, Dako, Glostrup, Denmark Positive control Lymph node Placenta Testicle
Pannoramic Desk Scanner Pannoramic Viewer 3DHistech, Budapest, HUNGARY
5%, 10% and then in 10% increments )
Multiply (0-300) Nuclear staining Cytoplasmic staining when present
– Mann-Whitney Test for non paired samples – Wilcoxon test for paired samples
for Windows, Version 22.0. Armonk, NY: IBM Corp.,
Mean Std. Deviation non neoplastic 20 26 LOW GRADE 21 33, HIGH GRADE 47 31 TREATED 61 52 CRPC 59 53 NECA 104 70 p<0,001 p=0,009 p=0.031
Low grade High grade
Non-neoplastic
CRPC NECA
Mean Std. Deviation non neoplastic 28 33 LOW GRADE 42 34 HIGH GRADE 46 30 TREATED 94 42 CRPC 80 31 NECA 87 48
p<0.001
Mean
LOW GRADE 76 26 HIGH GRADE 72 20 TREATED 78 20 CRPC 81 13 NECA 60 31
DNMT2 Cytoplasmic staining
Low grade High grade
Non-neoplastic
CRPC treated
Mean Std. Deviation non neoplastic 19 17 LOW GRADE 30 39 HIGH GRADE 18 21 TREATED 21 41 CRPC 110 70 NECA 160 37 p<0.001
CRPC Low grade High grade NECA
times during prostate cancer progression
PCa progression with significant elevation in NECa
ablation therapy
castration resistance
DNMT3b may be a promising therapeutic target for CRPC Both DNMT1 and DNMT3b may be promising therapeutic targets for NECa
DNMT3L across the spectrum of prostate cancer progression
deregulation
University of Patras)
MADCC)
Oncology, MADCC)
Oncology, MADCC)
MADCC)
the University of Patras via “K. Karatheodori” program
421, Research Committee of the University of Patras