1. Novel blood and tissue Biomarkers for Breast and Prostate Cancers
Elena Klenova (
NHS Consultants
Chandra Sekharan
John Corr
Phil Murray
Ian Seddon
University of Essex team
Metodi Metodiev
Yukti Gupta
Viv D’Arcy
Dawn Farrar
Jay Mani
Naresh Pore
Anu Apte
Adhip Mandal
Zubair Cheema
Research Nurses
Kathy Rooke
Andrea Appleby
Louise Robb
Devi Basu
Essex Biomedical
Sciences Institute (EBSI)

2. PLAN OF THE TALK
Protein BORIS -a potential breast and prostate blood and tissue cancer biomarker.
BORIS belongs to the Cancer-Testis antigen family which is normally present only in the testis but aberrantly activated in various malignancies.
(Loukinov et al 2002, PNAS; Klenova et al 2002, Sem Cancer Biol)
2. Identification and characterization of new blood biomarkers for early diagnosis, prognosis and treatment of breast and prostate cancer

3. BORIS – a novel breast tumour tissue biomarker
Immunohistochemistry analysis
IRS = Immuno Reactivity Score (calculated to assess the levels of a protein)
Western blot analysis

4. BORIS levels are higher in breast tumour tissues with different phenotypes than in normal breast tissues
BR – breast reduction
PP – paired peripheral
All – all tumours
High levels of BORIS correlate with high levels of the Oestrogen and Progesterone Receptors
D’Arcy et al, British Journal of Cancer (2008).

5. BORIS – a novel prostate tumour tissue biomarker
BORIS staining on Prostate Tumour Tissue Sections
IRS= 9
IRS= 8
IRS= 3
Non Tumour
BPH
Prostate Tumour
IRS=0
IRS=0
Secondary only
Cheema, Hari et al in preparation (2013).

6. BORIS in Tissues: Clinical Correlations
Gleason Score
Tumour stage

7. Clinical Correlation: BORIS and Androgen Receptor in prostate tumors
Mean BORIS IRS

8. Blood Leukocyte Biomarker
BORIS – a novel blood biomarker detected in leukocytes of breast cancer patients
Cancer Patient T
Interaction between
tumour and immune system
Change in protein profiles
Blood Leukocyte Biomarker

9. Immunohistochemistry analysis
Western blot analysis
IRS = Immuno Reactivity Score (calculated to assess the levels of a protein)
D'Arcy V et al. Clin Cancer Res 2006;12:

10. BORIS is detected in the leukocytes of breast cancer patients
(a) phenotypes of matching tumours (b) BORIS levels increase with the tumour size
D'Arcy V et al. Clin Cancer Res 2006;12:

11. BORIS is detected in leukocytes of prostate cancer patients
Healthy Donor
IRS= 8
IRS= 6
IRS= 4
IRS= 0
Secondary Only

12. BORIS in Leukocytes: Clinical Correlations

13. Assays for BORIS detection from whole blood (“point of care device”)
  ELISA
(Enzyme linked immunosorbent assay)
Immunohistological methods.
Real-time RT-PCR
Immunofluorometric methods
(flow cytometry)

14. Identification of novel blood leukocyte-based biomarkers
Cancer Patient T
Interaction between
tumour and immune system
Change in protein profiles
Blood Leukocyte Biomarker

15. Identification and characterization of new blood biomarkers for (i) early diagnosis and prognosis or breast and prostate cancers, and (ii) treatment monitoring of breast cancer
Patient cohorts
Breast and prostate cancer patients and healthy donors.
Metastatic breast cancer patients undergoing chemo - and endocrine therapies.

16. The Search for Novel Blood Biomarkers for Breast and Prostate Cancers
Blood collection
Plasma
White Blood Cells
Cell pellets X
RBC
2 D gel electrophoresis
Cell pellets
Protein identification by Mass Spectrometry
Protein extraction
Candidate Validation
Massive parallel protein
profiling using the Orbitrap Velos instrument

17. Protein Identification/ Candidate Blood Biomarkers for metastatic cancer treatment
78 Candidates
11 No Ids ( unknown? Novel biomarkers?)
7 Bacteria proteins
Several biomarkers are currently being validated.

18. Conclusions
Protein BORIS has been identified and evaluated as a novel potential breast and prostate blood and tissue cancer biomarker.
Work in progress to further evaluate BORIS for the use in the clinic.
More new blood biomarkers for early diagnosis, prognosis and treatment of breast and prostate cancer are being identified using high throughput genomics and proteomics techniques.
Work in progress to validate these biomarkers in relation to clinical outcomes.
3. In some cases biomarkers = DRUG TARGETS ( new dimensions)