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Innsbruck Medical University, Austria (principal investigator: Prof. dr. Zoran Culig)

Prostate cancer development is at least in part caused by pro-inflammatory cytokines such as interleukin (IL)-6, -4, and -8. In this project, we will use a number of prostate cancer cell lines to identify members of the Bcl-2 family and other inhibitors of apoptosis whose expression is commonly disregulated in prostate cancer. Genomic and in silico proteomics approaches will be implemented to analyze profiles of molecules regulated by those cytokines in prostate cancer models and clinical specimens.

Anti-IL-6 treatment with a targeted antibody has been demonstrated in various cellular models; in this project, we will combine that treatment with either anti-androgenic, cytotoxic, or anti-angiogenetic therapies in vitro and in vivo. Molecular targets for these combined treatments will be identified on the basis of validated array analyses. Preclinical studies should serve as a basis for clinical trials with combination therapies.

Institute for Molecular Medicine Finland (principal investigator: Prof. dr. Olli Kallioniemi)

We have developed a unique ultra-high density cell-microarray system for RNA interference screening, allowing up to 10,000 siRNA transfections per experiment. Cell phenotypes resulting from the knockdown of specific genes are registered using multi-parametric assays. The aim of this project will be to identify and validate genes whose knockdown sensitize prostate cancer cells to androgen deprivation and induce apoptosis.

We will bioinformatically analyze the hit genes by e.g. exploring their (co-) expression profiles in clinical prostate cancer specimens. The significant challenge in large-scale screens is the functional validation of the findings, which takes time and effort, and requires expertise on specific signaling pathways. In PRO-NEST, we will enable other network members to analyze or produce high-throughput RNAi data and will then resort to their expertise in validating specific hits in their domains of expertise.

University of Tampere, Finland (principal investigator: Prof. dr. Tapio Visakorpi)

Our preliminary data on non-coding RNAs (ncRNAs) indicate that they are involved also in the tumorigenesis of prostate cancer. In this project, we aim to elucidate the role of ncRNA in prostate cancer. The emphasis will be on evaluating the ncRNA, especially microRNAs (miRNAs), as diagnostic/prognostic tools as well as novel therapy targets. The expression, genetic and epigenetic alterations, as well as function of miRNAs in prostate cancer will be studied.

Also, the targets of the miRNAs will be investigated. The analyses will be based on using population based clinical materials collected by the UTA, and other partners, and on models.

The project is part of the Translational Prostate Cancer Research Programme (TPCRP, conducted at UTA and Tampere University Hospital), creating an excellent platform for translational training of the Fellows. The project will also be integral part of the EU-FP7 programme ProspeR.

4. Gene expression profiling of prostate cells with a selected panel of quantitative nucleic acid biomarkers
and
5. Proteomic multiplex assays for prostate tissue lysates and blood

University of Turku, Finland (principal investigator: Prof. dr. Kim Pettersson)

The two PhD projects within the Dept of Biotechnology of U.Turku relate to the set-up of high quality, fully quantitative and specific methodologies for determination of either 1) gene or 2) protein expression in patients specimens represented by prostate cells derived from prostate biopsies, voided urine, bone marrow aspirates or from the circulation or by forms secreted or leaked into the blood. Sample sets for the initial marker validation consists primarily from biopsies of more than 300 post radical prostatectomy patients from U.Turku University Hospital to yield both RNA stabilized preparations and protein lysates of tumor and control areas. These samples have been collected since 2000 and also serum and blood preparations (PaxGen) samples are available for each patient.

The marker candidates tested so far as well as future candidates will be selected from biomarkers suggested to be of diagnostic and/or prognostic value as appearing in the literature and/or by the partners of this proposal or through the FP6 P-Mark project. In this proposal close collaborations will be with EMC, PHILIPS, RUNMC and ULUND, who can validate their nucleic acid and protein marker candidates using the high performance and robust technologies and assay concepts of U.Turku.

CERBM, France (principal investigator: Prof. dr. Bohdan Wasylyk)

We have identified small molecules, such as XRP44X, that could be useful for prostate cancer treatment, thereby moving our research from the bench towards the bedside. XRP44X targets the Ras-Net signalling pathway and inhibits prostate cancer metastasis in animals. We are screening for small molecules that target TTLL12, a protein over-expressed in prostate cancer metastasis.

The project is to characterize these molecules in cell and animal models of prostate cancer, in order to understand their molecular mechanisms of action. These chemical inhibitors will be used as tools that are complementary to conventional methods (expression vectors and siRNAs), which act over a longer time frame, and may not be suitable to study dynamic changes in signaling pathways. The studies in animals will contribute to the pre-clinical analyses required to validate the molecules for clinical studies in humans.

Forschungzentrum Karlsruhe GmbH, Germany (principal investigator: Prof. dr. Andrew Cato)

We have identified Hag-2 and Hag-3 (otherwise known as Agr2 and Agr3) as over-expressed proteins in prostate cancer. Hag-2 and Hag-3 are the human homologues of the secreted Xenopus laevis proteins XAG-1/2 (AGR-1/2) expressed in the cement gland, an ectodermal organ in the head associated with dorsoanterior ectodermal cell fate determination during early development. The physiological roles of Hag-2 and Hag-3 in mammalian cells are unknown. Hag-2 is reported in mucous secreting cells and endocrine organs.

The aim of this project is to determine whether these two proteins can be used as markers for prostate cancer and to analyze their role in prostate cancer development using conventional knock-down or over-expression studies. We will pay particular attention to the secretory nature of these proteins and determine whether they are involved in autocrine or paracrine responses in prostate tumorigenesis.

University of Münster, Germany (principal investigator: Prof. dr. Axel Semjonow)

UMUNS has a large collection of surgical tissue specimens, as well as serum and plasma obtained from the same PCa patient population at radical prostatectomy or prior to prostate biopsy. The detailed clinical characteristics as well as long-term follow up (5 to 12 years) of these patients are available at UMUNS. A Tissue Microarray (TMA) of more than 950 of these patients has been established, 570 of the pts. received no additional therapy to surgery.

Recent investigations of this population consisted of IHC techniques (i.e. biomarkers on 8p-21-22 (NKX3.1, LOXL2) and biomarkers of the epidermal mesenchymal transition (E-cadherin, SNAIL, N-cadherin)) and characterisation of cytokeratin expression (CK5/6, CK14, CK19, CK8/18). In addition, biomarkers of proliferation or apoptosis have been determined using MIB1 and XIAP.

In a prospective study, (more than 200 radical prostatectomies are performed at UMUNS annually) selected entire prostates will be investigated using SNP-Microarrays (e.g. LOH-tool Affymetrix) generating the genomic characterisation of normal tissue in comparison to various prostate cancer foci flanked by IHC validation of the already investigated biomarkers. UMUNS has a long standing experience with differential evaluation of multifocal prostate cancers in native prostate tissue.

Based on traditional histological grading for each individual focus, each focus will be characterised by the tissue biomarkers described above and compared to genomic loss or gain of chromosomal aberrations. The next step is to include tissue biomarkers from EMC and ULUND in this study.

RUNMC, the Netherlands (principal investigator: Prof. dr. Jack Schalken)

The role of cell adhesion molecules in cancer is well documented. In epithelial derived cancers impaired E-cadherin function and a switch to the mesenchymal subtypes of cadherins was reported. The cadherins interact with the actin cytoskeleton via catenins (α, β & γ).

We recently identified a protein FAM 110A by two hybrid screening which interacts with ?-catenin. Over-expression results in increased motility and decreased cell adhesion. Interestingly, a family member FAM110B is mapped to a region on chromosome 8 that is amplified in PrCa. This is associated with over-expression of FAM110B. We hypothesize that FAM110B is an α -catenin binding protein that is over-expressed in PrCa through gene amplification. The over-expression leads to impaired cadherin function.

This project will study the biological functional role of FAM110A- and -B in prostate cancer. We will profile over- and under-expressing cells and perform bioinformatic analyses (Kallioniemi), to better understand the downstream mechanisms affected by FAM110B. The clinical relevance will be studied in prostate cancer specimens and urine. Thus we will find evidence for the significance of FAM110B in prostate cancer and determine its clinical utility.

RUNMC, the Netherlands (principal investigator: Prof. dr. Jack Schalken)

RUNMC has a unique collection of surgical specimens obtained from patients that were progressive under endocrine therapy, called castration resistant PCa. The samples will be used for gene expression profiling using Affymetrix arrays. This platform is well validated and operational at RUNMC.

The bioinformatic analyses will be done at the CMBI (Centre for Biomolecular Informatics, RUNMC; head Prof. dr. G. Vriend). Thus, all technology platforms and bioinformatics infrastructure and expertise is available. Real time PCR assays will be developed for candidate biomarkers and tested on tissue specimens, mononuclear cell fractions and urine. The biomaterials are available including a detailed annotation (clinical follow up etc.). It is important that comparison with the recently established biomarkers PCA3 and TMPRSS2-erg gene fusions can be made, so as to have a rational decision tree for selection of biomarkers. The selected biomarkers will be developed on an TMA (target mediated amplification) platform. Initial clinical validation can be done and is feasible. RUNMC has a track record of successful target validation and early clinical testing (e.g. PCA3 and TMPRSS2-erg).

Erasmus MC, the Netherlands (principal investigators: Prof. dr. Chris Bangma, Dr. Guido Jenster, Dr. Theo Luider, Dr. Arno van Leenders)

A series of novel proteins and peptides have been revealed in our ongoing efforts in the FP6 P-Mark project that are differentially present in serum from men without and with PCa. These proteins and peptides may serve as serum biomarkers for diagnosis and prognosis, as expression of some of these is correlated to disease progression. The next step is to unequivocally confirm the differential expression of the peptides in an extended set of patient serum samples, available in a well characterised biorepository with long clinical follow-up. For this purpose, we will use TripleQuad Mass Spectrometry (MS) in combination with the selective reaction monitoring (SRM) method. The TripleQuad first scans for the masses of preselected series of peptides of interest, after which it will perform MS/MS to obtain transitions that are specific for the peptides of interest. This method is fast, selective and highly sensitive, and makes it possible to check the quantity of many different peptides in many different patient samples.

Based on the outcome of this evaluation combined with datamining using information technology tools developed in EMC, selected promising candidate biomarkers will enter the Biomarker Validation subprogramme. Specific immunoassays for these candidates will be developed in collaboration with U.Turku, or existing assays will be utilized if available in order to test large series of patient serum samples for biomarker expression in the various PCa disease stages to determine diagnostic and prognostic power. In addition, candidate biomarker expression will be analyzed by immunohistochemistry on Tissue MicroArrays (TMA) and other formalin-fixed, paraffin-embedded (FFPE) prostate (cancer) tissues including biopsies to study in which cell type and to what extend the candidate biomarker is produced, revealing whether marker expression, particularly in biopsies, has prognostic value.

At the EMC and other partners, large collections (over 1000) of serum matched with FFPE tissues (from radical prostatectomies) and diagnostic biopsies, are available to instigate statistical sound study designs. For this research project we will closely collaborate with various partners including U.Turku, PHILIPS and UMUNS.

Erasmus MC, the Netherlands (principal investigators: Prof. dr. Jan Trapman, Dr. Guido Jenster)

1M Illumina SNP and Affymetrix EXON array analyses of prostate cancer patient samples and xenografts, revealed a wealth of cancer-related DNA and RNA changes. Of particular interest to us are the borders of chromosomal deletions and amplifications and the genes located on these breakpoints. Therefore, we will integrate SNP and EXON array data of the same 100 patient samples to identify DNA breakpoints resulting in aberrant gene transcripts.

In addition, DNA and RNA fusions will be identified using state of the art Solexa Paired-End High Throughput Sequencing in which both ends of RNAs and DNA fragments are sequenced. This allows for identification of fragments in which the ends do not match according to the human genome sequence revealing potential fusion genes.

Candidate fusion genes will be further studied for their occurrence rate and role in cancer initiation and progression.

Lund University, Sweden (principal investigators: Prof. dr. Anders Bjartell, Prof. dr. Hans Lilja)

High-throughput large scale analyses of PCa specimens by immunohistochemistry (IHC) have become common after the introduction of tissue microarrays and recently developed scanning procedures of processed slides using automated image analysis, web-based reading and storage of images. However, the evaluation of tissue biomarkers is still hampered by the lack of reliable methods to quantify immunostaining intensity.

The conventional enzymatic methods, which currently are used for IHC were developed long ago and are unable to provide quantitative detection signals corresponding to levels/amounts of antigen present in a tissue section. Improved low-end detection performance combined with enhanced linearity/dynamics relating to specific immunodetection of molecular markers in tissue sections can be obtained employing time-resolved fluorescence (TRF) microscopy, a technology utilizing the uniquely slow-decay of certain fluorescent labels to avoid non-specific interference from auto-fluorescence.

Access to a workstation for TRF microscopy in Malmö has allowed us to develop novel and highly sensitive methods using primary antibodies labeled with fluorescent europium or terbium chelates. The student will use established PCa biomarkers (PSA, androgen receptor) to investigate the possible advantages with TRF microscopy for immunostaining intensity quantification in FFPE tissue sections from core biopsies and radical prostatectomy specimens obtained from unique population-based cohorts (Malmö Preventive Medicine and Malmö Diet and Cancer cohorts) and from a prospective collection comprising more than 700 patients in the P-Mark project. Next, novel biomarkers in development by ULUND and other partners can be validated with this technology.

University of Bern, Switserland (principal investigators: Prof. dr. Marco Cecchini, Prof. dr. George Thalmann)

The University of Bern has established an innovative method able to dissect the stromal compartment-specific from the cancer cell-specific gene expression in bone metastases in vivo. In collaboration with the Swiss Institute of Bioinformatics (Epalinges, Switzerland) we created a computational mask, which discriminates gene expression between cancer cell (human) and stroma (mouse host) to identify interactions between these cell types in chimeric human-mouse cancer models.

Stromal- and epithelial-specific gene expression have been analysed according to this method in xenografts of human, osteinductive CaP cell lines (C4-2B-4 and VCaP) in immuno-compromised mouse bone. Seventy-seven genes have been found to be up-regulated in the bone stromal compartment at least 8-fold, as compared to sham-operated, control bone stroma. Of these, 23 are extracellular matrix proteins, 12 are signalling molecules, 6 are molecules involved in cell-cell or cell-matrix interaction and 7 are secreted proteases and related inhibitors. Included are also annotated genes of unknown function (12) and non-annotated genes (5).

We propose to analyze by the same method the stromal gene expression response induced by bone xenografts of the human, osteolytic CaP cell line PC3. This analysis should potentially highlight different signalling mechanisms explaining the different response, osteolytic or osteoblastic, in cancer bone metastasis.

University of Cambridge, United Kingdom (principal investigator: Dr. Ian Mills)

The androgen receptor (AR) is the archetypal lineage survival oncogene in prostate cancer based on the hormone responsiveness of early stage disease. The subsequent discovery of other lineage survival oncogenes such as the ETS factors and the realization that these transcription factors may interact and be co-dependent raises the possibility that other transcription factors could be equally or more significant for the development of aggressive prostate cancer.

Existing information on gene targets will be applied to siRNA screens in collaboration with Olli Kalloniemi and further ChIP experiments to compare the overlap in gene targets between the AR and the major candidate co-dependent transcription factors (FoxA1, ETS, GATA-2, Oct1). The development and characterization of induced pluripotent stem (iPS) cells by other members of PRO-NEST will refine our understanding of the critical conserved gene targets and expression signatures for prostate cancer and tissue development.

University of Newcastle upon Tyne, United Kingdom (principal investigator: Prof. dr. Craig Robson)

Availability of prostate stem cells will greatly facilitate our understanding of prostate development and the progression to cancer. Stem cell research has been transformed by the Yamanaka group?s discovery that adult skin cells can acquire properties of embryonic stem cells through simple genetic manipulation. Problems that remain to be addressed include confirmation that induced pluripotent stem (iPS) cells possess the inherent ability of embryonic stem cells to produce the complete repertoire of adult cell types; establishing if differences exist for iPS cells derived from various tissues; establishing methods of generating iPS cells without involving permanent genetic modification with associated risk of malignant transformation.

We will establish novel prostate iPS cell clones and perform comparative analysis with conventional skin iPS cells (we have successfully generated germ cell iPS cells). Undifferentiated embryonic cell transcription factor-1 (UTF-1) will be employed as a pluripotent selection marker. UTF-1 mRNA transcripts are only detected in native embryonic cells and embryonic stem cells derived from adult tissues. Assays will be conducted to validate differentiation potential and to assay tumorigenic potential (unlikely since our protocol excludes c-myc retrovirus in iPS generation).

We will rigorously compare the properties of iPS cells derived from skin with "physiological" prostate tissue-derived iPS cells through proteomic profiling, RNA microarray analysis and expression of stem cell markers. Findings will be validated in an orthotopic prostate xenograft mouse model.

University of Oxford, United Kingdom (principal investigator: Prof. dr. Freddie Hamdy)

Recent data have shown that a global disruption of microRNA occurs in prostate cancer. The profile of this disruption appears specific to the extent of disease, treatment with androgen deprivation and function of the androgen receptor. More specifically loss of mir-146a occurs in hormone refractory prostate cancer cell lines. Re-expression of this molecule reduces invasion, proliferation and the development of bone metastases in the same cell lines.

Here we will focus on the development of bone metastases in advanced disease by profiling a series of paired cell lines and tumor samples. The latter include primary and metastatic specimens, whilst the former will include cell lines with low/high metastatic sub-clones, hormone dependent/independent variants, including PC-3, PC3M, PC3-LN4, LNCaP, C4-2, LNCaP-LN3.

We will determine global microRNA expression using Agilent microRNA microarrays. Differentially expressed microRNA molecules will be individually analyzed by measuring the phenotype (invasion and metastatic assay) of "knock in? (transfection with pre-miRs (Applied Biosystems, UK)) and "knock out? (siRNA to microRNA, Applied Biosystems, UK) cell lines models.

We will also investigate these microRNA molecules (using QrtPCR and FISH) in several large series of prostate cancer samples (stratified for primary, metastases and various treatment strategies with known clinical outcomes.

University of Oxford, United Kingdom (principal investigator: Prof. dr. Freddie Hamdy)

Chromosomal rearrangements play a causal role in cancer. These arrangements are of clinical interest and could be used to adapt therapy. Recently, gene fusions involving the prostate-specific gene transmembrane protease, serine 2 (TMPRSS2) and members of the erythroblastosis virus E26 transforming sequence (ETS) family of transcription factors were identified in up to 60% of PCa. The prognosis significance of this fusion is still matter of debate, but the fusion is detectable in blood using FISH method applied on isolated PCa circulating cells.

This method is currently not clinically applicable and is not a quantitative technique. We have therefore examined in a preliminary study the quantitative detection of the fusion transcript using RT-PCR techniques in advanced PCa cases. The fusion could be detected in up to 10 fusion positive PCa cells on 100000 fusion negative PCa cells, and was also found in the peripheral blood of 3 on 12 advanced PCa cases.

The objectives of this PhD project are to study if detection of TMPRSS2 fusions in peripheral blood can be used as a detection and/or monitoring tool for PCa, to measure its performance and compare the clinical outcomes according to fusion status, and to explore its possible complementary value to urine tests for similar fusions and PCA3 expression. The presence of the fusion will be investigated on blood using RT PCR and on the prostate biopsies using RT PCR and FISH methods. The quantification of the fusion will also be monitored after androgen withdrawal for patients having sequential blood samples taken and correlated to PSA and clinical outcomes.

University of Oxford, United Kingdom (principal investigator: Prof. dr. Freddie Hamdy)

This project is closely linked to PhD project 5 from UOXF and moves towards the development of tailored therapy guided by biomarker-based diagnostics.

Gene fusions of TMPRSS2 and members of the ETS family are present in tumor tissue from up to 60% of the PCa patients. Tailored therapy targeted at this chromosomal rearrangement may improve the current rate for the treatment of both localised and disseminated PCa. The expression of TMPRSS2:ETS fusion is up-regulated by androgens and blocked by anti-androgens, but the impact of other anti cancerous drugs is unknown.

In this project, the impact of various drugs on PCa cell lines growth according to their TMPRSS2:ETS status will be studied. VCaP and LNCaP cells, PCa cells that are harvesting TMPRSS2:ERG and TMPRSS2:ETV1 respectively, will be used. The TMPRSS2 fusions will be knocked-down using siRNA. The transcriptome profile will be compared between normal and knocked-down cells using microarray technology. According to the pathways that are likely to be involved from microarray data, the cell growth and the level of fusion transcripts will be compared using anti-cancerous drugs. Drugs efficient in PCa like docetaxel, mitoxantrone, and estramustine will be investigated, as well as others according to the pathways that will be found to be activated. Potential candidates may be ERbeta agonists (resveratrol and genistein), zoledronic acid and various EGFR, VEGF or PDGF inhibitors.

University of York (principal investigators: Prof. dr. Norman Maitland, Dr. Anne Collins)

Prostate epithelium in normal, premalignant and malignant states exists as a hierarchy of cells in different differentiated states. Gene expression profiling of fractionated cell populations has indicated that some genes are obligatorily silenced during this differentiation process.

While most gene expression profiles are from either cell lines or total cell / micro-dissected epithelium) and therefore dominated by the genes for highly expressed secreted luminal cell products, our studies have focused on more basal cell types from primary tissues, wherein lie the stem cells for both malignant and non-malignant tissues.

This project will further develop methods to carry out expression profiling and fate determination on small highly homogeneous cell populations from primary human cells, to ascertain the effects of reactivation of silenced genes in these populations and to explore the molecular basis of the observed gene silencing at the chromosomal level.

Philips Electronics Nederland B.V., The Netherlands (principal investigators: Dr. Ralf Hoffmann, Dr. Edwin Romijn)

We propose to develop prostate cancer cell lines that stably express a reporter gene for the in vivo imaging of in vivo applied cells in either optical and/or nuclear imaging systems. These cellular reporter systems allow the easy and sensitive monitoring of the fate of grafted cells in living immuno-compromised animals (e.g., to follow the development of metastases from a primary tumor locations).

Furthermore, we will extend such cell lines by stably transfecting them with a Tet-regulated activator protein (rtTA) to make them ready as an expression system for markers of prostate cancer under an rtTA regulated and inducible promoter.

By the use of these systems we will test the influence and the function of the expression of target proteins that have been proposed to function in the formation of metastatic growth or having a role in the transformation from hormone-dependent to hormone-independent cancer growth.

Philips Electronics Nederland B.V., The Netherlands (principal investigators: Dr. Ralf Hoffmann, Dr. Edwin Romijn)

Philips Research Europe is working on protein biomarker discovery and protein biomarker validation in the area of oncology, and more specifically in the area of PCa that will serve as content of the technology platforms capable of measuring biomarkers that are also being developed within Philips Research. Protein biomarker discovery and protein biomarker validation, especially when done in serum or plasma, is limited by mainly protein dynamic range, and overall sensitivity and often lack of high-quality antibodies against the target of interest.

The proposed post-doc research project will work on the development of novel approaches to deal with these aspects in relation to protein biomarker evaluation and validation, which overcomes part of the mentioned limitations. Multiple Reaction Monitoring (MRM) mass spectrometry coupled to multi-dimensional separation techniques and the use of stable isotopes for quantification purposes, in combination with bioinformatics tools to predict fragmentation behavior in a Triple Quadrupole Mass Spectrometer and retention behavior in multi-dimensional separations of biomarker candidates will be developed.

Biomarker candidates will come from several ongoing discovery activities making use of severalbackToTop technical approaches, ranging from COFRADIC, glycoproteomics, and 4-dimensional separation techniques. Scientific exchange at this level will primarily be with EMC and UOXF. Subsequent development of clinically applicable assays for selected candidates biomarkers for further validation in the Biomarker Validation subprogramme will be done in collaboration with U.Turku.

SERVICEXS, The Netherlands (principal investigators: Dr. Wilbert van Workum, Dr. Bart Janssen)

SNP-chip analyses (CNV), Affymetrix exon array analysis and next-generation sequencing technology (Illumina Genome Analyzer) of prostate cancer samples revealed many novel cancer-related DNA and RNA changes. An assay to detect rare sequence variants in the presence of an overwhelming population of wild-type sequences would provide a powerful molecular diagnostic tool with several applications, e.g. early tumor detection, and identification of tumor cells in patients in relapse.

Pyrophosphorolysis-activated polymerization (PAP) for allele-specific amplification has been shown to selectively detect single-nucleotide variant in the presence of >109 copies of wild-type sequence. Within this experienced researcher project we propose to develop PAP assays that could be useful to monitor the efficacy of prostate cancer treatment both on the DNA and RNA level, the latter including fusion transcripts and splice variants.

Lund University, Sweden (principal investigators: Dr. Yvonne Ceder, Prof. dr. Anders Bjartell, Prof. dr. Hans Lilja)

Using an existing real time quantitative RT-PCR assay for non-invasive detection of a selected microRNA (miRNA) in serum samples, the fellow will analyse levels of a pre-selected miRNA in patient serum samples, to determine if there is a correlation to the levels in the prostate and to diagnosis. This part of the project will examine miRNA in the context of (i) whether the selected miRNA identifies disease that needs to be treated (and also therefore indolent disease that does not require radical treatment), (ii) whether the selected miRNA identifies very aggressive forms of disease (require early radical and multiple treatments).

If significant discriminatory potential can be observed, we will continue to investigate whether the selected miRNA also have prognostic potential. This will be possible to investigate due to access to a major blood biorepository, Malmö Preventive Medicine (MPM). This was a very large project that took place 1974-1986 with a follow-up of over 25 years, in total 21,277 men in certain age cohorts, participated in the MPM project, representing 74% of eligible men. The MPM project was focusing on cardiovascular and metabolic risk factors, and all participants completed an extensive baseline questionnaire, including family history of serious illnesss, lifestyle factors, various cardiovascular, gastrointestinal, genitourinary signs or symptoms, and a large panel of biochemical tests measured in blood. Consistent with current national guidelines in Sweden, participants were never recommended to undergo early screening for PCa. Of the participating men, 462 have later been diagnosed with PCa (median delay 18 years).