Training and validation of a novel 4-miRNA ratio model (MiCaP) for prediction of postoperative outcome in prostate cancer patients.

Background: New molecular biomarkers for prostate cancer (PC) prognosis are urgently needed. Ratio-based models are attractive, as they require no additional normalization. Here, we train and independently validate a novel 4-miRNA prognostic ratio model for PC. Patients and methods: By genome-wide miRNA expression profiling of PC tissue samples from 123 men who underwent radical prostatectomy (RP) (PCA123, training cohort), we identified six top candidate prognostic miRNAs and systematically tested their ability to predict postoperative biochemical recurrence (BCR). The best miRNA-based prognostic ratio model (MiCaP) was validated in two independent cohorts (PCA352 and PCA476) including >800 RP patients in total. Clinical end points were BCR and prostate cancer-specific survival (CSS). The prognostic potential of MiCaP was assessed by univariate and multivariate Cox-regression analyses and Kaplan-Meier analyses. Results: We identified a 4-miRNA ratio model, MiCaP (miR-23a-3p×miR-10b-5p)/(miR-133a×miR-374b-5p), that predicted time to BCR independently of routine clinicopathologic variables in the training cohort (PCA123) and was successfully validated in two independent RP cohorts. In addition, MiCaP was a significant predictor of CSS in univariate analysis [HR 3.35 (95% CI 1.34 - 8.35), P = 0.0096] and in multivariate analysis [HR 2.43 (95% CI 1.45-4.07), P = 0.0210]. As proof-of-principle, we also analyzed MiCaP in plasma samples from 111 RP patients. A high MiCaP score in plasma was significantly associated with BCR (P = 0.0036, Kaplan-Meier analysis). Limitations include low mortality rates (CSS: 5.4%). Conclusions: We identified a novel 4-miRNA ratio model (MiCaP) with significant independent prognostic value in three RP cohorts, indicating promising potential to improve PC risk stratification.

Prognostic significance of aberrantly silenced ANPEP expression in prostate cancer.

BACKGROUND: Novel biomarkers for prostate cancer (PC) are urgently needed. This study investigates the expression, epigenetic regulation, and prognostic potential of ANPEP in PC. METHODS: Aminopeptidase N (APN; encoded by ANPEP) expression was analysed by immunohistochemistry using tissue microarrays representing 267 radical prostatectomy (RP) and 111 conservatively treated (CT) PC patients. Clinical end points were recurrence-free survival (RFS) and cancer-specific survival (CSS), respectively. The ANPEP promoter methylation levels were determined by bisulphite sequencing or MethyLight analysis in 278 nonmalignant and PC tissue samples, and in cell lines. RESULTS: The APN expression was significantly downregulated in PC compared with nonmalignant prostate tissue samples. Aberrant promoter hypermethylation was frequently observed in PC tissue samples, and 5-aza-2'-deoxycytidine induced ANPEP expression in three hypermethylated prostate cell lines, suggesting epigenetic silencing. Negative APN immunoreactivity was significantly associated with short RFS and short CSS in the RP and CT cohort, respectively, independently of routine clinicopathological predictors. Combining APN with a known angiogenesis marker (vascular endothelial growth factor or microvessel density) improved risk prediction significantly in both cohorts. CONCLUSION: Our results suggest negative APN immunoreactivity as a new independent adverse prognostic factor for patients with clinically localised PC and, furthermore, that epigenetic mechanisms are involved in silencing of ANPEP in PC.

miR-145 induces caspase-dependent and -independent cell death in urothelial cancer cell lines with targeting of an expression signature present in Ta bladder tumors.

Downregulation of miR-145 in a variety of cancers suggests a possible tumor suppressor function for this microRNA. Here, we show that miR-145 expression is reduced in bladder cancer and urothelial carcinoma in situ, compared with normal urothelium, using transcription profiling and in situ hybridization. Ectopic expression of miR-145 induced extensive apoptosis in urothelial carcinoma cell lines (T24 and SW780) as characterized by caspase activation, nuclear condensation and fragmentation, cellular shrinkage, and detachment. However, cell death also proceeded upon caspase inhibition by the pharmacological inhibitor zVAD-fmk and ectopic expression of anti-apoptotic Bcl-2, indicating the activation of an alternative caspase-independent death pathway. Microarray analysis of transcript levels in T24 cells, before the onset of cell death, showed destabilization of mRNAs enriched for miR-145 7mer target sites. Among these, direct targeting of CBFB, PPP3CA, and CLINT1 was confirmed by a luciferase reporter assay. Notably, a 22-gene signature targeted on enforced miR-145 expression in T24 cells was significantly (P<0.00003) upregulated in 55 Ta bladder tumors with concomitant reduction of miR-145. Our data indicate that reduction in miR-145 expression may provide bladder cancer cells with a selective advantage by inhibition of cell death otherwise triggered in malignant cells.

Genome-wide analysis of allelic imbalance in prostate cancer using the Affymetrix 50K SNP mapping array.

Prostate cancer (PCa) is the most commonly diagnosed non-cutaneous cancer in male subjects in Western countries. The widespread use of prostate-specific antigen (PSA) has increased the detection of this cancer form in earlier stages. Moreover, it has increased the need for new diagnostic procedures to be developed for patient stratification based on risk of progression. We analysed laser-microdissected prostate tumour tissue from 43 patients with histologically verified PCa, using the new high-resolution Affymetrix Mapping 50K single-nucleotide polymorphism array. The results showed six major loss of heterozygosity regions at chromosomes 6q14-16, 8p23-11, 10q23, 13q13-21 and 16q21-24 and a novel region at chromosome 21q22.2, all of which reveal concomitant copy number loss. Tumour development was further characterised by numerous novel genomic regions almost exclusively showing copy number loss. However, tumour progression towards a metastatic stage, as well as poor differentiation, was identified by specific patterns of copy number gains of genomic regions located at chromosomes 8q, 1q, 3q and 7q. Androgen ablation therapy was further characterised by copy gain at chromosomes 2p and 10q. In conclusion, patterns of allelic imbalance were discovered in PCa, consisting allelic loss as an early event in tumour development, and distinct patterns of allelic amplification related to tumour progression and poor differentiation.

To grow old. From a socio-medical epidemiological intervention study among old citizens of Copenhagen.

A cross-sectional, epidemiological, sociomedical survey, followed by intervention, was conducted with the participation of 585 men and women in the 75, 80 and 85 year-old age groups living in the city of Copenhagen. The main objectives of the study were to describe the health and social conditions of the elderly people, to register the physical and social unmet needs and then intervene to relieve these needs, to assess the effect of such an intervention through a follow-up study, and to demonstrate possible risk factors. Age, marital status, recent hospitalisation were shown to be connected with the functional condition. The quality of life study showed that health, functional ability to stay independent, and housing conditions are most important factors when growing old. The socio-medical intervention did not show any effect with regard to mortality, hospitalisation, institutionalisation, subjective health and economy, loneliness, quality of life and functional ability. The most striking risk markers in relation to death were: low systolic blood pressure among women, subjective poor health, low score on the ladder-scale used to quantify quality of life, and impaired functional capacity in nine out of ten partial functions. All together, it is concluded that if we want to improve the living conditions of the elderly we should reserve the efforts for old people at risk and enable as many as possible to stay independent.

[The geriatric clinical picture in a department of internal medicine]. / Det geriatriske sygdomsbillede i en intern-medicinsk afdeling.

In order to assess the proportion of geriatric patients in a medical department, all of the inpatients admitted to the Medical Department of Hørsholm Hospital during the period 19.3.1987-15.4.1987 were assessed in view of the geriatric clinical picture. The criteria for assessment as a geriatric patient were established in advance and included: Presence of several medical problems inhibition of functions and social problems and an estimated need for prolonged hospitalization to solve these problems, including rehabilitation. Out of 191 inpatients, 15 (8%) were considered to be geriatric patients. These 8% were all aged 60 years or more, were hospitalized for an average of 23 days and used 21% of the total number of bed days. The remaining patients aged more than 60 years were hospitalized for an average of nine days and comprised 47% of the total number of inpatients and used 49% of the bed days. The geriatric patients differed, in addition, from the other patients aged over 60 years in that 85% compared with 58% were readmitted within a period of two years. No difference in the mortality was observed during a period of two years. This investigation supports previous suppositions about the proportion of geriatric patients and indicates that assessment as to whether a geriatric patient is concerned or not may be made already during the first 24 hours in hospital.

[Cooperation concerning admission to and discharge of elderly people from the hospital. 3. Functional capacity, self-assessed health and quality of life]. / Samarbejde om gamle menneskers sygehusindlaeggelse og udskrivelse. 3. Funktionsevne, selvvurderet helbred og livskvalitet.

As part of a total project concerning description and effects of the coordinating efforts of home nurses in connection with admission to and discharge from hospital of persons over the age of 65 years, the functional capacity and the subjective parameters: quality of life, self-assessed health and loneliness were registered on discharge and two months later. In order to assess the effect of intervention, the intervention group was compared with a control group. No differences in the functional capacity could be demonstrated on discharge or after two months. No differences could be demonstrated in the subjective parameters either. The effect of the home nurses' efforts in the total project could be measured as a reduction in the number of institution-days in the intervention group. These results suggest that, in further attempts to reduce the risk of complications in recently discharged elderly patients, further assistance of the functional capacity should be added to the forms of intervention described.