Prostate cancer: early detection, monitoring and prognosis:

Essex Biomedical Sciences Institute (EBSI)

Elena Klenova, University of Essexthe search for a magic molecule

Most Common Types of Cancers in Men

info.cancerresearchuk.org/.../males/index.htm

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 biomarkerDRE (Digital Rectal Examination)

• Treatment: Active Surveillance, Surgery, radiation therapy, hormonal therapy, chemotherapy

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

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.

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.

3. Treatment and its monitoring

Androgen-dependent Androgen-Independent

Why Advanced Prostate Cancer Becomes Resistant to Androgen-Deprivation Therapy?

New therapies?

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.

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

N C

1 2 3 4 5 6 7 8 9 10 11Structure ofBORIS protein

DNA-binding region

BORIS- the solution to all three issues?

In press

• 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

• 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)

Immuno Histochemistry

IRS= 9 IRS= 8 IRS= 3

Secondary onlyIRS=0 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

BORIS in prostate tumour tissues: positive correlations with poor prognosis

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

Clinical Correlation: BORIS and AR in prostate tumors

Are the AR mRNA and protein levels affected by BORIS

over-expression(+) or BORIS knock down(-)?

Plate cells

Mix BORIS DNA/BORIS siRNA withtransfection 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

BORIS positively regulated the AR geneAR mRNA

BORIS mRNA

AR and BORIS protein analysis

BORIS positively regulated the AR gene in prostate cancer cells

ARGene

ARGene

ARGene

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!

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.

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.

Thank you! Elena Fabiola Georgia Dawn Jay Ola Zubair Anu Adele

Funded byColchester Hospital University NHSColchester Catalyst CharityUniversity of Essex

Clinical collaboratorsMr. J. Corr, Mr. R. Casey

AcknowledgementsDr. I. Seddon, Sister Lucy PowelHistopathology staff at C.G.H.

Svetlana Hulkar Krista Yukti

Essex Biomedical Sciences Institute (EBSI)

Thank you!

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

BORIS expression in prostate cell lines

BORIS in prostate tumours: immunofluorescence staining