Prostate cancer: early detection, monitoring and prognosis:

1. Essex Biomedical Sciences Institute (EBSI) Prostate cancer: early detection, monitoring and prognosis: the search for a magic molecule Elena Klenova, University of Essex

2. Most Common Types of Cancers in Men info.cancerresearchuk.org/.../males/index.htm

3. What is Prostate Cancer? • Prostate Gland is a part of male reproductive system • Risk Factors: Age, Family History, Diet • Major Symptoms: Lower back pain, slow/ painful urination, blood in urine or semen • Diagnosis: PSA blood test (Prostate Specific Antigen)- prostate cancer biomarker DRE (Digital Rectal Examination) • Treatment: Active Surveillance, Surgery, radiation therapy, hormonal therapy, chemotherapy

4. Prostate cancer - Facts • Most cases of prostate cancer develop in older men (over the age of 65) • 35 000 cases diagnosed / year (UK) • 10 000 deaths / year (UK) • Second most common cause of cancer death • More than 200 men are diagnosed with Prostate Cancer at the Urology Department (CHUFT) each year; 281 new cases in 2011

5. MAJOR ISSUES – match Prostate Cancer UK there are three priority areas • Identifying men at highest risk of developing clinically significant prostate cancer. • 2. Distinguishing aggressive prostate cancer from indolent disease. Understanding how aggressive prostate cancers develop. • 3. Finding new, targeted treatments for advanced prostate cancer.

6. 1-2 Prostate cancer development and detection Slow growing and takes years before it is detected. Does not always need to be treated. Only few patients have an aggressive form of prostate cancer that grows much more quickly. Currently it is hard to detect which form of cancer a patient has, which can make prostate cancer treatment decisions more difficult.

7. 3. Treatment and its monitoring Why Advanced Prostate Cancer Becomes Resistant to Androgen-Deprivation Therapy? New therapies? Androgen-Independent Androgen-dependent

8. The search for “MAGIC” molecules specific for prostate cancer disease • Useful as BIOMARKERS ( disease –specific markers) for diagnosis and prognosis of a condition. • Help to understand how aggressive cancer develop and distinguish. • Useful as targets for specific anti-prostate cancer therapies.

9. 1 2 3 4 5 6 7 8 9 10 11 BORIS- the solution to all three issues? • BORIS:Brother Of the Regulator of Imprinted Sites • Member of CTA (Cancer Testis Antigen) family - only • expressed in the testis, but not in other normal tissues. • Aberrantly expressed in different malignancies, including • prostate cancer. Zinc Fingers Structure of BORIS protein N C DNA-binding region

10. In press

11. AIM OF THE STUDY: investigate the association of BORIS with prostate cancer • Investigate BORIS expression in prostate tissues obtained from patients with prostate cancer • Correlate the BORIS results with clinical information(Gleason Score, Tumour stage, Androgen Receptor status) • Can BORIS be the magic molecule ?

12. Recruitment Process ( coordinated by Zubair Cheema) • Patients with histological diagnosis of Prostate Cancer (Sister Lucy Powell) • Patient Discussion and Consent pre op • Surgery at C.G.H. (Mr. Corr and Mr. Casey) • Tissue Transfer to Pathology • Tissue Assessment (Dr. Ian Seddon) • Tissue frozen at stored at UoE

13. Immuno Histochemistry BORIS staining on Prostate Tumor Tissue Sections IRS= 8 IRS= 3 IRS= 9 Non Tumor Prostate Tumor BPH IRS=0 Secondary only IRS=0 No BORIS staining was observed in non-tumor and BPH tissue sections

14. BORIS in prostate tumour tissues: positive correlations with poor prognosis

15. Androgen Receptor (AR) • Activated by binding of androgens (e.g. Testosterone). • Essential for normal and abnormal growth of prostate. • Higher levels of AR are often detected in more aggressive prostate cancers Happy cell Production of proteins to support growth of prostate cells mRNA

16. Clinical Correlation: BORIS and AR in prostate tumors

17. Are the AR mRNA and protein levels affected by BORIS over-expression(+) or BORIS knock down(-)? Western blot assay for protein analysis qPCR for mRNA analysis Plate cells Add mix to the cells Mix BORIS DNA/ BORIS siRNA with transfection reagent To measure AR and BORIS levels Incubate for 48 hrs and assay

18. AR mRNA AR and BORIS protein analysis BORIS positively regulated the AR gene BORIS mRNA

19. BORIS positively regulated the AR gene in prostate cancer cells AR Gene AR Gene AR Gene

20. Role of BORIS in the development of more aggressive prostate cancer? Androgen Receptor (AR) is key for prostate cancer progression and then development of the independence from androgens. Advanced Prostate Cancer Difficult to cure! Androgen-Independent Androgen-dependent

21. Conclusions • BORIS is present in prostate tumor tissues but absent in normal and Benign Prostate Hyperplasia (non-tumour) tissues: BORIS is a marker for prostate cancers. • A positive correlation exists between BORIS IRS and Gleason Scoring /Tumor Stage: BORIS is a marker for poor prognosis of prostate cancer. • BORIS is an activator or the Androgen Receptor gene: BORIS is associated with the development of more aggressive prostate cancers. • BORIS is present only in cancer cells and therefore is a very attractive target for anti-tumour therapy.

22. BORIS - the “MAGIC” molecules specific for prostate cancer disease? • Useful as BIOMARKERS ( disease –specific markers) for diagnosis and prognosis of a condition. • Help to understand how aggressive cancer develop and distinguish. • Useful as targets for specific anti-prostate cancer therapies.

23. Thank you! Elena Fabiola Georgia Dawn Jay Ola Zubair Anu Adele Svetlana Hulkar Krista Yukti Clinical collaborators Mr. J. Corr, Mr. R. Casey Acknowledgements Dr. I. Seddon, Sister Lucy Powel Histopathology staff at C.G.H. Funded by Colchester Hospital University NHS Colchester Catalyst Charity University of Essex

24. Essex Biomedical Sciences Institute (EBSI) Thank you!

25. Western Blotting-BORIS protein levels in prostate cancer cell lines and tumours

26. BORIS expression in prostate cell lines

27. BORIS in prostate tumours: immunofluorescence staining