- Email: [email protected]
203 Urine biomarkers for prostate cancer
203
Urine biomarkers for prostate cancer Eur Urol Suppl 2014;13;e203
Print! Print!
Yazbek Hanna M. 1 , Clark J. 2 , Hurst R.2 , Brewer D.2 , McCarthy F.3 , Mills R.1 , Rochester M. 1 , Edwards D.2 , Cooper C.2 1 Norfolk
and Norwich University Hospital, Dept. of Urology, Norwich, United Kingdom, 2 University of East Anglia, Dept. of Biological
Sciences, Norwich, United Kingdom, 3 Institue of Cancer Research London, Dept. of Research, Norwich, United Kingdom INTRODUCTION & OBJECTIVES: Serum PSA testing has well-recognised deficiencies as a prostate-cancer-specific diagnostic and screening test, deficiencies which have clinical implications. Thus, Improved biomarkers are therefore urgently required to more accurately diagnose prostate cancer, and to distinguish aggressive from non-aggressive disease. Due to the interconnection of the prostatic ducts and the urinary tract, urine-based tests for prostate cancer have the potential to sample biological materials from multiple prostate cancer foci in one assay overcoming the problem of innate heterogeneity and multifocality of the disease. In this project we are working as a part of a Movember funded global consortium that aims to discover a robust set of molecular markers that will be of both diagnostic and prognostic value. MATERIAL & METHODS: First catch urine was collected from 500 men after digital rectal examination. Patients were divided into 6 groups: 1) negative for cancer: PSA100 & clinically malignant; 3-5) Low Intermediate and High risk of metastasis as substratified per NICE criteria, 6) post prostatectomy (no DRE). Methods have been optimised to harvest several biological fractions for analysis in collaborating laboratories by proteomics, metabolomics, DNA-methylation, cell and exosomal RNA. The focus of our group is on the analysis of the exosomal RNA fraction. RESULTS: RNA yields and gene analysis comparisons between pre- and post-prostatectomy samples indicated prostatic origin of the material. Pilot NanoString gene analysis using the nCounter human cancer 236-gene assay showed expression of 189 out of 236 genes, which confirm the utility of these analyses. NanoString and RT-PCR probes were compared for 4? genes but were not always in agreement. NanoString expression analysis of 52 genes has been performed on 193 samples. Preliminary analysis shows over expression of Hepsin, UPK2, PCA3 and HOXC6 in cancer patients compared to benign controls. Further data analysis will be available at presentation. CONCLUSIONS: Good yields of prostatic exosomal RNA can be harvested from urine allowing multiple gene analysis using both RT-PCR and NanoString. Preliminary analysis shows over expression at least 4 gene probes in cancer samples.
Urine biomarkers for prostate cancer Eur Urol Suppl 2014;13;e203
Print! Print!
Yazbek Hanna M. 1 , Clark J. 2 , Hurst R.2 , Brewer D.2 , McCarthy F.3 , Mills R.1 , Rochester M. 1 , Edwards D.2 , Cooper C.2 1 Norfolk
and Norwich University Hospital, Dept. of Urology, Norwich, United Kingdom, 2 University of East Anglia, Dept. of Biological
Sciences, Norwich, United Kingdom, 3 Institue of Cancer Research London, Dept. of Research, Norwich, United Kingdom INTRODUCTION & OBJECTIVES: Serum PSA testing has well-recognised deficiencies as a prostate-cancer-specific diagnostic and screening test, deficiencies which have clinical implications. Thus, Improved biomarkers are therefore urgently required to more accurately diagnose prostate cancer, and to distinguish aggressive from non-aggressive disease. Due to the interconnection of the prostatic ducts and the urinary tract, urine-based tests for prostate cancer have the potential to sample biological materials from multiple prostate cancer foci in one assay overcoming the problem of innate heterogeneity and multifocality of the disease. In this project we are working as a part of a Movember funded global consortium that aims to discover a robust set of molecular markers that will be of both diagnostic and prognostic value. MATERIAL & METHODS: First catch urine was collected from 500 men after digital rectal examination. Patients were divided into 6 groups: 1) negative for cancer: PSA100 & clinically malignant; 3-5) Low Intermediate and High risk of metastasis as substratified per NICE criteria, 6) post prostatectomy (no DRE). Methods have been optimised to harvest several biological fractions for analysis in collaborating laboratories by proteomics, metabolomics, DNA-methylation, cell and exosomal RNA. The focus of our group is on the analysis of the exosomal RNA fraction. RESULTS: RNA yields and gene analysis comparisons between pre- and post-prostatectomy samples indicated prostatic origin of the material. Pilot NanoString gene analysis using the nCounter human cancer 236-gene assay showed expression of 189 out of 236 genes, which confirm the utility of these analyses. NanoString and RT-PCR probes were compared for 4? genes but were not always in agreement. NanoString expression analysis of 52 genes has been performed on 193 samples. Preliminary analysis shows over expression of Hepsin, UPK2, PCA3 and HOXC6 in cancer patients compared to benign controls. Further data analysis will be available at presentation. CONCLUSIONS: Good yields of prostatic exosomal RNA can be harvested from urine allowing multiple gene analysis using both RT-PCR and NanoString. Preliminary analysis shows over expression at least 4 gene probes in cancer samples.