Personalized prostate cancer prevention and management

Approved 07/09/2020, Comitato Etico Territoriale Interaziendale AOU Maggiore della Carità (Corso Mazzini 18, Novara, 28100, Italy; +39 (0)3213733081; comitatoetico@maggioreosp.novara.it), ref: Prot. N°968/CE 07/09/2020, integrations Prot. N°263/CE, 10/03/2021 and Prot. N°250/CE, 21/03/2023 specific for the NW ancillary study

Blood sampling is performed at baseline and: every 6 months, for at least three times, in men without any prostate cancer suspicion; at each recall in men with prostate cancer suspicion; before prostatectomy and at each follow-up up to three years after radical prostatectomy in men who undergo surgery. Plasma only is isolated at every time point after baseline.
1. Serum, plasma and PBMCs are isolated within 1 hour after blood sampling at baseline using centrifugation and Ficoll gradient separation
2. Plasma is isolated at every time point after baseline using centrifugation of EDTA K2 tubes at 2500 rpm for 10 min at 4°C, repeated twice
3. Serum is isolated at baseline using centrifugation of serum-specific tubes at 2500 rpm for 10 min at 4°C
4. PBMCs are isolated at baseline using Ficoll gradient separation
5. Urine supernatant and pellet are isolated at baseline or before fusion biopsy using centrifugation at 15000 rpm for 30 min at 4°C
6. PSA concentration is measured using Elecsys total PSA kit IVD at baseline and at each follow-up time point
7. Plasma circulating levels of miR-103a-3p, miR-5100, and let-7a-5p are measured using RT-qPCR and Taqman assays at baseline and after prostatectomy in men diagnosed with prostate cancer and treated surgically
8. Plasma circulating sphingolipids are measured using targeted lipidomics via mass spectrometry at baseline and after prostatectomy in men diagnosed with prostate cancer and treated surgically
9. Plasma circulating proteome is measured using the Olink Explore HT platform at baseline, after prostatectomy in men diagnosed with prostate cancer and treated surgically, and after 6 months in men enrolled in the NW ancillary study
10. Transcriptomic profile of PBMCs is measured using total RNA sequencing at baseline
11. Transcriptomic profile of fixed prostate tissue is measured using total RNA sequencing in men with intermediate-risk prostate tumors (ISUP grade 2–3)
12. Urinary microbiota is measured using Pathochip arrays (Agilent) starting from DNA and RNA isolated from urine sediments at baseline or before fusion biopsy
13. Lifestyle factors including physical activity, BMI, diet, smoking and alcohol consumption are measured using self-reported questionnaires at baseline
14. Familial history, past prostate biopsies and concurrent pathologies are measured using self-reported questionnaires at baseline
15. PSA levels and digital rectal examination outcomes are measured using clinical records and urological visit reports at baseline and at each follow-up time point
16. Histological and clinical information for prostate tumors is recorded using pathology reports and clinical records at each follow-up time point
17. Electronic Case Report Forms (eCRF) including questionnaires, urological visit reports, PSA dosage and lab analyses are uploaded to a password-protected online database (https://www.keyform.eu) at each follow-up time point

These measures are carried out in the ancillary study only:
1. Sedentary behavior is measured using the Global Physical Activity Questionnaire (GPAQ) at baseline, 6 months, and 1 year (for NW group only)
2. Physical activity in occupational, transport-related, and leisure-time contexts is measured using the Global Physical Activity Questionnaire (GPAQ) at baseline, 6 months, and 1 year (for NW group only)
3. Weekly level of moderate to vigorous physical activity (MVPA) is measured using metabolic equivalents (METs) derived from GPAQ data at baseline, 6 months, and 1 year (for NW group only)
4. Psycho-physical health and quality of life are measured using the SF-12 questionnaire at baseline, 6 months, and 1 year (for NW group only)
5. Height, weight, and hip and waist circumferences are measured using anthropometric assessment at baseline, 6 months, and 1 year (for NW group only)
6. Body mass index (BMI) is calculated from height and weight measurements at baseline, 6 months, and 1 year (for NW group only)
7. Blood pressure, O2 saturation level, and resting heart rate are measured using clinical assessment at baseline, 6 months, and 1 year (for NW group only)
8. Upper limb strength is measured using the Handgrip Test at baseline, 6 months, and 1 year (for NW group only)
9. Dynamic balance, mobility, and fall risk are measured using the Timed-Up-and-Go Test at baseline, 6 months, and 1 year (for NW group only)
10. Lower limb strength and endurance are measured using the 30’’ Sit-to-Stand Test at baseline, 6 months, and 1 year (for NW group only)
11. Aerobic capacity and endurance are measured using the 6 Minute Walking Test (6WT) at baseline, 6 months, and 1 year (for NW group only)
12. Rating of perceived exertion (RPE) is measured using subjective self-assessment during the 6 Minute Walking Test at baseline, 6 months, and 1 year (for NW group only)
13. Body roundness index (BRI) is calculated using waist circumference and height measurements at baseline, 6 months, and 1 year (for NW group only)
14. A Body Shape Index (ABSI) is calculated using waist circumference, BMI, and height measurements at baseline, 6 months, and 1 year (for NW group only)
15. Conicity index is calculated using waist circumference, weight, and height measurements at baseline, 6 months, and 1 year (for NW group only)
16. Plasma circulating proteome is measured using the Olink Explore HT platform at baseline and after 6 months (for NW group only)