1. “Clinical and molecular effects of dietary supplements in patients with ASAP and/or multifocal HGPIN: a randomized double-blinded placebo controlled phase I-II study" Francesco Soria Clinica urologica Università degli Studi di Torino

2. Introduction Globocan 2012 PCa is an ideal target for chemoprevention due to its long natural history and it's high incidence Disease incidence changes according to environmental factors and race Dietary influence on PCa is well documented Selenium, lycopene, green tea catechins

3. Introduction Lopez JI, BJU Inter 2007 Men with multifocal HGPIN have a 20- 40% of chance of PCa at rebiopsy Even higher risk is present for ASAP Multifocal HGPIN and ASAP are well documented precursors and predictors of PCa

4. Matherials and methods Double blind Phase I-II randomized placebo controlled study 3 times a day Selenium-methionin (title 0.512%, 3.58 mg/cps) Lycopene (title 15%, 77.78 mg/cps) Green tea catechins (title 65%, 210 mg/cps) Phase I Verification of pharmacological stability, tolerability and blood level concentration Phase II Comparison of PCa incidence at rebiopsy between supplementation and placebo groups

5. Results: Baseline characteristics Variable All patients N= 60 Placebo arm N= 30 Treatment arm N= 30 p Age mean (SD) 63.3 (7.0) 62.6 (8.2) 64.1 (5.7)0.41 PSA mean (SD) 6.3 (3.2) 6.5 (3.7) 6.2 (2.6)0.74 DRE + mean (%) 34.8 26.7 43.30.27 Mean biopsy cores (SD) 14.9 (3.3) 15.1 (4.4) 14.8 (1.8)0.73 ASAP overall (%) monofocal multifocal 32 (53.3) 24 8 17 11 6 15 13 2 0.62 HGPIN (%) 18 (30.0) 9 91.00 HGPIN+ASAP (%) 10 (16.7) 4 60.73

6. Results: Phase I No alterations up to three years were found both under standard and accelerated conditions for green tea extract and seleniomethionine Seleniomethionine in accelerated conditionsGreen tea extract in accelerated conditions Contrarily lycopene extract was found particularly sensitive to light, atmospheric oxygen and temperature Lycopene in accelerated conditions HPLC method allowed to highlight the presence of lycopene in the plasma of the enrolled subjects

7. Results: Efficacy Variable All Patients 6 months N (%) PLACEBO 6 months N 26 (%) ACTIVE TREAT 6 months N 27 (%) P Prostate cancer Overall Gleason 6 Gleason 7 Gleason ≥ 8 13 (24.5) 6 7 0 3 (5.7) 1 2 0 10 (18.9) 5 5 0 0,053 Mean biopsy cores (SD) Mean n of positive cores (HGPIN + ASAP) 16.0 (3.0)15.0 (2.0)15.7 (3.3)1,00 ASAP overall monofocal multifocal 981981 330330 651651 0.47 HGPIN 4221.00 HGPIN+ASAP 2111.00 NEGATIVE 2516956 PSA (SD) 6.2 (4.6)6.5 (4.3)6.0 (4.9)0.74 DRE + (%) 36.5 24.048.1 0.28

8. Results: microRNA expression profiling (Agilent platform) To test whether treatment could modify the microRNA expression profile of HGPIN/ASAP tissue microenviroment 16 samples 8 patients (4 for group) Tissue areas adiacent to HGPIN/ASAP at first biopsy and negative or adjacent to PCa at second biopsy A consistent group of microRNAs, overexpressed in PCa vs non cancer stroma upregulated in the supplementation group MicroRNA suppressing the proliferation, invasion, and migration of PCa downregulated in the supplementation group

9. Conclusions The supplementation promoted rather than prevented progression to PCa both from clinical and molecular results The risk of chemoprevention turning into "chemopromotion" when using the assessed dietary supplements in PCa high risk patients with HGPIN and/or ASAP should be acknowledged Carefulness in patient prescriptions and awareness in future studies should be kept

10. Grazie! “Clinical and molecular effects of dietary supplements in patients with ASAP and/or multifocal HGPIN: a randomized double-blinded placebo controlled phase I-II study"