TMPRSS2 rs12329760 variant as a prognostic marker for prostate cancer progression.

Objective To study the expressions of transmembrane protease serine 2-E26 transformation specific (TMPRSS2-ETS) fusion gene and ETS related gene (ERG), ETS variant 1 (ETV1), phosphatase and tensin homolog deleted on chromosome 10 (PTEN), androgen receptor (AR) in prostate cancer. Methods A total of 50 cases of prostate cancer paraffin specimens and 30 cases of benign prostatic hyperplasia as the controls in the same period from the Third People's Hospital of Datong between December 2016 and October 2017 were collected. TMPRSS2-ERG, TMPRSS2-ETV1 and TMPRSS2-ETV4 gene fusion status were detected by using fluorescence in situ hybridization. Immunohistochemistry was used to detect the expressions of ERG, ETV1, PTEN and AR protein. Results In prostate cancer tissues, TMPRSS2-ETS fusion gene positive rate was 70% (35/50), and TMPRSS2-ETS fusion gene was not detected in benign prostatic hyperplasia. The expression of TMPRSS2-ETS fusion gene in prostate cancer patients with different age, serum prostate specific antigen level and whether distant metastasis had no statistically significant differences (all P > 0.05). The expression of TMPRSS2-ETS fusion gene in Gleason score > 7 patients was higher than that in Gleason score≤7 patients (P 0.05). The expressions of TMPRSS2-ETS fusion gene and ERG in prostate cancer were positively correlated (r=0.302, P 0.05). Conclusions The expressions of TMPRSS2-ETS fusion gene and ERG, ETV1 protein are upregulated in prostate cancer, which may involve in the occurrence and development of prostate cancer. The detection of TMPRSS2-ETS fusion genes and ERG protein can provide a reference for the diagnosis of prostate cancer. Key words: Prostatic neoplasms; Transmembrane protease serine 2; E26 transformation specific; Gene fusion; Fluorescence in situ hybridization

Context:Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cellular entry is facilitated by transmembrane protease serine 2 (TMPRSS2), which is regulated by the androgen receptor (AR). Androgen deprivation therapy (ADT), widely used in prostate cancer treatment, may potentially modulate TMPRSS2 expression, affecting SARS-CoV-2 infection susceptibility and severity.Objective:To evaluate the impact of ADT on pulmonary TMPRSS2 expression in prostate cancer patients and analyze differences in expression patterns associated with specific ADT regimens.Design:We examined TMPRSS2 immunohistochemical expression in lung tissue from 20 consecutive autopsy cases of men with prostate cancer (6 receiving ADT at time of death), compared with non-ADT prostate cancer patients and age-matched women controls. Histoscores were calculated by assessing percentage and intensity of pneumocyte TMPRSS2 expression.Results:Prostate cancer patients receiving ADT showed significantly reduced pulmonary TMPRSS2 expression compared to non-ADT patients (mean histoscores: 152.7 vs. 225.0, p=0.037) and age-matched women controls (mean histoscores: 152.7 vs. 238.0, p=0.024). Direct AR antagonists (apalutamide, bicalutamide) produced more pronounced TMPRSS2 suppression than GnRH modulators or androgen biosynthesis inhibitors. No significant correlation was observed between TMPRSS2 expression and Gleason score, PSA levels, or underlying lung pathology.Conclusion:Our findings demonstrate that ADT significantly reduces pulmonary TMPRSS2 expression, with direct AR antagonists showing the strongest effect. This suggests a potential mechanistic explanation for differential COVID-19 susceptibility and provides rationale for investigating AR-targeted therapies as potential protective interventions against SARS-CoV-2 infection severity.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cellular entry is facilitated by transmembrane protease serine 2 (TMPRSS2), which is regulated by the androgen receptor (AR). Androgen deprivation therapy (ADT), widely used in prostate cancer treatment, may potentially modulate TMPRSS2 expression, affecting SARS-CoV-2 infection susceptibility and severity. We evaluated the impact of ADT on pulmonary TMPRSS2 expression in prostate cancer patients and analyzed differences in expression patterns associated with specific ADT regimens. We examined TMPRSS2 immunohistochemical expression in lung tissue from 20 consecutive autopsy cases of men with prostate cancer (6 receiving ADT at time of death), compared with non-ADT prostate cancer patients and age-matched women controls. Histoscores were calculated by assessing the percentage and intensity of pneumocyte TMPRSS2 expression. Prostate cancer patients receiving ADT showed significantly reduced pulmonary TMPRSS2 expression compared to non-ADT patients (mean histoscores: 152.7 vs. 225.0, p = 0.037) and age-matched women controls (mean histoscores: 152.7 vs. 238.0, p = 0.024). Direct AR antagonists (apalutamide, bicalutamide) produced greater TMPRSS2 suppression than Gonadotropin-Releasing Hormone modulators or androgen biosynthesis inhibitors. No significant correlation was observed between the TMPRSS2 expression and Gleason score, PSA levels, or underlying lung pathology. Our findings demonstrate that ADT significantly reduces pulmonary TMPRSS2 expression, with direct AR antagonists showing the strongest effect. This suggests a potential mechanistic explanation for differential COVID-19 susceptibility and provides a rationale for investigating AR-targeted therapies as potential protective interventions against SARS-CoV-2 infection severity.

Estrone sulfate (E 1S), a prognosis marker for tumor aggressiveness in prostate cancer (PCa)

The transcriptional activity of androgen receptor (AR) is modulated by coregulators that have a significant influence on a number of functional properties of AR, including the ligand specificity as well as the DNA binding capacity. Because androgens and AR have pivotal roles in the development and

To explore the correlation between plasma level of lncRNA TUG1 with PSA level, Gleason grading and tumor node metastasis (TNM) stage of prostate cancer (PCa) patients. This study aims to evaluate the potential diagnostic and prognostic values of TUG1 in PCa. Plasma level of TUG1 in 70 PCa patients and 70 healthy controls was determined using the quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Correlation between plasma level of TUG1 with PSA level, Gleason grading and TNM stage of PCa patients was analyzed. The potential diagnostic value of TUG1 in PCa was explored by introducing the receiver operating characteristic (ROC) curve. The survival analysis of PCa patients undergoing radical prostatectomy was conducted using the Kaplan-Meier method and the log-rank test. Finally, the regulatory effects of TUG1 on in vitro proliferative and migratory abilities of PCa cells were examined by the cell counting kit-8 (CCK-8) and the transwell assay, respectively. QRT-PCR data revealed a higher plasma level of TUG1 in PCa patients than in those of healthy controls. In particular, PCa patients with stage III+IV had a higher level of TUG1 relative to those with stage I+II. Moreover, the TUG1 level was higher in PCa patients with a higher PSA level (≥ 10 ng/mL), Gleason grading (≥ 7) or TNM stage (N1, M1). There was no significant correlation between the plasma level of TUG1 and the age of PCa patients. The ROC and Kaplan-Meier curves indicated the diagnostic and prognostic values of TUG1 in PCa. The overexpression of TUG1 markedly accelerated PCa cells to proliferate and migrate. The plasma level of TUG1 is upregulated in PCa patients and is correlated to PSA level, Gleason grading and TNM stage of PCa. TUG1 exerts certain diagnostic and prognostic values in PCa. The overexpression of TUG1 markedly accelerates PCa cells to proliferate and migrate.

e17069 Background: Classical androgen receptor (AR) target is the new “tumor-specific” antigen for prostate cancer vaccine development to delay progression and castration-resistant disease. HLA-A2-restricted AR ligand-binding domain (LBD) has been demonstrated to stimulate cytotoxic T lymphocytes (CTL) in HLA-A2+ prostate cancer patients. Although the prevalence of HLA-A2 serotype and LBD-specific CD8+ T cells among Caucasians are known, their frequencies in the Taiwan patients have not been established. In this study, we performed HLA-A2 genotyping, HLA-A2+ LNCaP stimulation of autologous T cell, and CTL tumor cell cytotoxicity from 25 castration-sensitive prostate cancer patients using blood samples collected from a single hospital center. Methods: 20ml whole blood samples are collected in EDTA tube from non-metastatic, castration-sensitive prostate cancer patients. PBMCs are subjected to HLA-A2 allele sequencing, separated by CD3 magnetic bead column, and expanded and activated ex vivo using CD3/CD8 polymeric beads. Autologous T cells are then isolated by CD8 magnetic bead column, stimulated by HLA-A2+ lentivirus-transduced LNCaP (AR+), and targeted against transduced (HLA-A2+) and non-transduced (HLA-A2-) LNCaP (AR+) by Chromium-51 Release Assay. We identified and correlated their frequencies with patient’s age at diagnosis, Gleason grades, PSA level, and family history. Results: A total of 25 blood samples were tested and analyzed for HLA-A2 genotype with 44% (11/25) positive for the HLA-A2 [28% (7/25) heterozygotes, 16% (4/25) homozygotes]. Among the 11 HLA-A2+ patients, 72% (8/11) were HLA-A*02:07 allele, 100% (11/11) CD8+ T cells were successfully stimulated with HLA-A2+ transduced LNCaP (AR+) and 45% (5/11) patients demonstrated HLA-A2+ AR-specific CTL tumor cell lysis. No clinical association with patient age (54-86 y.o.), Gleason grades (6-9), PSA level (3.84-62.8 ng/ml), and family history (1/25). Conclusions: In this single hospital center study, we observed a similar prevalence of HLA-A2 alleles in Taiwan prostate cancer patient cohort with a predominant HLA-A2*02:07 allele, and higher frequencies of HLA-A2-restricted CD8+ T cell stimulation and HLA-A2+ AR-specific tumor cell cytotoxicity. Our findings support the application of AR LBD as tumor antigen for prostate cancer vaccine development for Taiwan prostate cancer patients.

Prostate cancer and SARS-CoV-2: possible intersections through the TMPRSS2 pathway.

Prostate cancer is the second most common and the fifth leading cause of death by cancer in men worldwide now. The failure of androgen deprivation therapy (ADT) for prostate cancer caused by activated androgen receptor (AR) signaling pathways mostly found. The role of AR in growth and progression of prostate cancer is still unclear. Analysis of AR expression in prostate cancer has never been done in West Sumatera. This study aims to determine AR expression of prostate cancer and correlate with Gleason score and perineural invasion. A total of 56 prostate cancer from department of anatomical pathology in West Sumatera. Hematoxylin and eosin (HE) stained slides and paraffin blocks were retrieved. Slides of all cases were evaluated to review Gleason score, histopathological grading, WHO grade group based on ISUP 2014/WHO 2016 and perineural invasion. Androgen receptor immunohistochemistry (IHC) was applied on all cases. High AR expression was the mostly found (51,79%). The mostly prostate cancer is Gleason score 9 (44,64%), histopathological grading poorly differentiated/undifferentiated (76,78%), WHO grade group 5 (48,21%). Perineural invasion was noted in 39,29%. There was significant statistical correlation between AR expression and Gleason score, but no significant correlation with perineural invasion. AR expression is the important marker of prostate cancer progression.

-Objective To evaluate the diagnostic value of the ratio of transmembrane protease serine 2 (TMPRSS2) and ErS-related gene (ERG)'s fusion transcripts to PSA mRNA in prostate cancer. Methods A fluorescence quantitative reverse transcript (FQ-RT)-PCR method for detection of the ratio of TMPRSS2: ERG fusion transcripts to PSA mRNA was established. TMPRSS2: ERG transcripts and PSA mRNA in urine samples obtained from 99 patients with prostate cancer (PCa) and 107 patients with benign prostate hyperplasia (BPH) were detected by FQ-RT-PCR, and the ratio of TMPRSS2:ERG mRNA to PSA mRNA was calculated by 2-△Ct. The diagnostic efficacy of TMPRSS2: ERG in urine was evaluated. The correlations of the ratio of TMPRSS2: ERG mRNA to PSA mRNA with clinical and pathological grades were investigated. Results The ratio of TMPRSS2: ERG mRNA to PSA mRNA was significantly higher in PCa group than that in BPH group (0.101 vs 0.000, U=2 290.500, P 0.05). The area under curve (AUC) was 0.784 (95% CI:0.707～0.860). When cutoff value of the ratio of TMPBSS2: ERG mRNA to PSA mRNA was 3.0×105, the diagnostic sensitivity, specificity, positive predictive value, negative predictive value and accuracy rate were 75.8% (75/99), 94.4% (101/107), 92. 6% (75/81), 80.8% (101/125) and 85.4% (176/206), respectively. But the sensitivity was not correlated with PSA, Gleason grade and clinical stage (χ2=3.572, 2.278,3.707 respectively,P>0.05). Conclusions The ratio of TMPBSS2:ERG mRNA to PSA mRNA in the urine has certain sensitivity and specificity in the diagnosis of PCa. It can be used as a potential marker in the diagnosis of PCa. However, it can not be used as an index to monitor tumor progression or prognosis in PCa patients. Key words: Prostatic neoplasms; Serine endopeptidases; Proto-oncogene proteins c-ets; Oncogene peoteins; fusion; Reverse transcriptase polymerase chain reaction

BMP7: A New Bone Metastases Prevention?

Background: Prostate cancer (PCa) is the most commonly diagnosed cancer for men in the United States. Most of the PSA screening-detected PCa are slow-growing and not life-threatening. However, the majority of PSA-detected PCa receive aggressive treatment, which is often associated with significant morbidity. Clinical variables, such as Gleason Score (GS), PSA level, and tumor stage, are not sufficient to distinguish aggressive from indolent diseases at diagnosis. Biomarkers are urgently needed for more accurate prediction of individual tumor behavior and patient prognosis. DNA methylation has shown profound impact on cancer development and progression. In this study, we performed a genome-wide DNA methylation profiling of peripheral blood leukocytes to identify predictors of aggressive PCa. Methods: We performed leukocyte DNA methylation profiling in 140 GS=6 PCa patients and 147 GS ≥ 8 patients using the Illumina's Methylation 450K BeadChip. The differential methylated CpG position (DMP) was identified using the ChAMP analysis package implemented in R. Top differentially methylated CpG sites between less aggressive (GS=6) and more aggressive (GS≥ 8) patients were used to develop a signature to best cluster these two groups using semi-supervised recursively partitioned mixture modeling (SS-RPMM) algorithm. The white blood cell subtype composition was analyzed with RefbaseEWAS method. Ingenuity pathway analysis was used to find the enriched pathways that may determine aggressive prostate cancer. Results: Analyzing all the assayed CpG sites together, the mean methylation level appeared to be lower in GS ≥ 8 patients than in GS=6 patients. This global DNA hypomethylation was only significant at CpG sites located within gene body, shore, shelf, opensea and 3' UTR regions, but not in promoter and regions close to transcription start sites. There were many individual hypermethylated CpG sites in GS ≥ 8 patients that are mostly located in CpG islands within promoter and in the vicinity of transcription start sites. We then split the patients into a training (70 GS=6 and 73 GS ≥ 8 patients) and testing set (70 GS=6 and 74 GS ≥ 8 patients) to identify and validate a CpG methylation signature that best differentiates GS=6 and GS≥ 8 patients. The signature can identify a subgroup of aggressive PCa patients with higher risks of biochemical recurrence (HR=2.57, 95% Cl, 0.99-6.68). We also analyzed the white blood cell subtypes in patients and found that natural killer (NK) cell proportion is significantly decreased in patients who experienced biochemical recurrence (P=0.045), indicating weakened immune response. Conclusion: These data suggest that DNA methylation in peripheral blood leukocytes may serve as valuable biomarkers to supplement clinical variables to predict aggressive PCa at diagnosis and worse prognosis of PCa patients. Citation Format: Yuyan Han, Jia Li, Mutian Zhang, Jeri Kim, Xifeng Wu, Deqiang Sun, Jian Gu. Genome-wide DNA methylation profiling of peripheral blood leukocytes identifies predictors of aggressive prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3332.

5060 Background: The Nutritional Prevention of Cancer Trial demonstrated a 52% lower incidence of prostate cancer in its selenium (Se) arm. Consequently, this trial was designed to assess if Se supplementation would reduce the progression of prostate cancer in subjects on active surveillance for their disease. Methods: A Phase 2b randomized, double-blind placebo-controlled clinical trial was conducted in men with localized prostate cancer who had elected to forego therapy for active surveillance. Subjects <85 years of age with biopsy proven non- metastatic prostate cancer, prostate specific antigen (PSA) levels <50ng/ml, Gleason score <8, no other treatment for prostate cancer, and life expectancy ≥3 years were recruited. A total of 140 men were randomized to placebo (n=46), 200μg/day (n=47) or 800μg/day (n=47) Se p.o. (as selenized yeast) and followed every 3 months for up to 5 years. The primary endpoint was PSA velocity, a surrogate marker of prostate cancer progression. Serum levels of PSA and Se were determined at baseline and at 3-month intervals thereafter. Variance-stabilizing transformation ln(PSA+1) was carried out and PSA velocity (slope of ln(PSA+1)) was estimated using mixed effects regression models. Results: Following adjustment for age, body mass index, serum selenium levels, pack-years of smoking at baseline, duration on study, race, PSA analysis method, and Gleason score; percent changes in PSA velocity (90% CI) for 200 μg/day and 800 μg/day treatment groups vs. placebo were -2.85% (-7.19, 1.70) and -0.70% (-5.16, 3.98), respectively. Estimated mean PSA velocities and PSA doubling times for placebo, 200 μg/day and 800 μg/day treatment groups were 0.09, 0.07, and 0.09 ng/mL/yr; and 4.92, 5.86 and 5.34 years, respectively. Upon secondary analyses, PSA velocity was statistically significantly higher in smokers (vs. non-smokers) (percent change in PSA velocity and 90% CI: 0.09 (0.03, 0.14)). Conclusions: Selenium supplementation did not significantly affect PSA velocity in subjects with non-aggressive disease. Longitudinal analyses also suggest that active surveillance was an appropriate choice for these subjects. Cigarette smoking may influence prostate cancer progression and needs further investigation. This work was supported by PHS CA079080 and CA023074. No significant financial relationships to disclose.

Prostate cancer stem cells: molecular characterization for targeted therapy

TMPRSS2-Ets gene fusions were identified in prostate cancers where the promoter of transmembrane protease, serine 2 (TMPRSS2) fused with coding sequence of the erythroblastosis virus E26 (Ets) gene family members. TMPRSS2 is an androgen responsive transmembrane serine protease. Ets family members are oncogenic transcription factors that contain a highly conserved Ets DNA binding domain and an N-terminal regulatory domain.Fusion of these gene results in androgen dependent transcription of Ets factor in prostate tumor cells. The ERG is the most common fusion partner with TMPRSS2 promoter in prostate cancer patients. The high prevalence of these gene fusions, in particular TMPRSS2-ERG, makes them attractive as potential diagnostic and prognostic indicators, as well as making them a potential target for tailored therapies.This review focuses on the clinical and biological significance of TMPRSS2-ERG fusions and their role in PC development and progression.