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

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 1.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 Androgen-dependent Androgen-Independent Why Advanced Prostate Cancer Becomes Resistant to Androgen-Deprivation Therapy? New therapies?

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. 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 NC 1 2 3 4 5 6 7 8 9 10 11 Structure of BORIS protein DNA-binding region BORIS- the solution to all three issues?

10. In press

11. 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 ? AIM OF THE STUDY: investigate the association of BORIS with prostate cancer

12. 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 Recruitment Process ( coordinated by Zubair Cheema)

13. Immuno Histochemistry IRS= 9 IRS= 8 IRS= 3 Secondary only IRS=0 BORIS staining on Prostate Tumor Tissue Sections Non Tumor BPH Prostate Tumor 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 Production of proteins to support growth of prostate cells mRNA Happy cell

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(-)? Plate cells Mix BORIS DNA/ BORIS siRNA with transfection reagent Add mix to the cells Incubate for 48 hrs and assay To measure AR and BORIS levels qPCR for mRNA analysis Western blot assay for protein analysis

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

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

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

24. Essex Biomedical Sciences Institute (EBSI)

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