Benign Prostate Research Articles

Cell-autonomous complement C3 maintains prostate epithelial cell homeostasis and prevents malignant transformation

Abstract Intracellularly active complement C3 and C3a emerged as central regulators of normal immune cell function and proliferation via control of mTOR and glycolysis. Because increased glycolysis is a hallmark of many cancer cells, we hypothesized that augmented cell-intrinsic C3 activity may contribute to epithelial cell malignancy. However, while benign prostate epithelial cells (PECs) harbored expected intracellular C3/C3a storages, cancerous PECs, surprisingly, exhibited loss of C3/C3a. Further, C3/C3a reduction levels in PECs correlated inversely with worsening Gleason scores in patients, silencing of C3 in healthy PECs induced uncontrolled proliferation, and reconstitution of intracellular C3a in prostate cancer cell line DU145 normalized their hyper-proliferation. In line with an unexpected anti-proliferative role for C3a in PECs, C3ar−/−mice developed spontaneous hyperplasia of prostate tubular cells with age, treatment of DU145 tumors, implanted subcutaneously into the flanks of athymic mice, with C3a-expressing adenovirus significantly reduced tumor growth in vivo, and augmented C3 levels are associated with a better prognosis in patients with prostate and other types of epithelial cell cancers. RNA-sequencing of C3or C3aR-deficient human PECs identified intrinsic C3a as novel controller of several known PEC oncogenes, likely via up-stream impact on the c-MYC pathway. In sum, while intracellular C3 drives proliferation in immune cells, in (prostate) epithelial cells, cell-autonomous C3/C3a is a negative regulator of growths and, thus, represses oncogenic transformation. Such cell-specific intracellular C3/C3a activities should be taken into consideration when targeting this system therapeutically.

Efficacy and safety of Water Vapor Thermal Therapy in the treatment of benign prostate hyperplasia: a systematic review and single-arm Meta-analysis

BackgroundBenign prostatic hyperplasia (BPH) is a common chronic condition among men aged 50 or older, causing voiding and obstructive lower urinary tract symptoms. Water vapor thermal therapy (WVTT) using the Rezūm® system is a new minimally invasive surgical technique that is increasingly reported as a treatment for BPH.MethodsThe protocol was submitted to the PROSPERO registry. We searched PubMed, Web of Science, Embase, Cochrane Library and ClinicalTrials.gov up to July 29, 2022. Quality assessment was carried out by a 20-item checklist form prepared by the Institute of Health Economics (IHE). Double arcsine transformation was performed to stabilize the variance of the original ratio. When I2 > 50%, the random effect model was used to calculate the pooled parameters. Otherwise, the fixed effect model was used. 95% confidence intervals (CIs) were calculated. A leave-one-out sensitivity analysis was performed to evaluate the impact of each study on the pooled outcomes, and finally, Egger’s test was used to assess publication bias.ResultsA total of seven single-arm observational studies and one random controlled trial, including 1015 patients, were included. One year after WVTT, the International Prostate Symptom Score decreased by 11.37 (95% CI: -12.53, -10.21), the IPSS Quality of Life scale decreased by 2.59 (95% CI: -2.92, -2.26), the maximum urine flow rate increased by 5.26 ml/s (95% CI: 4.53, 5.99), and the postvoid residual decreased by 13.18 ml (95% CI: -24.32, -2.03). The most common complication was dysuria, with a pooled incidence of 21% (95% CI: 14%, 29%), and the second most common complication was hematuria, with a pooled incidence of 14% (95% CI: 10%, 18%). The pooled incidence of retreatment was 3% (95% CI: 2%, 5%).ConclusionsWVTT is an attractive alternative to medication or more invasive surgical procedures and can serve as first-line therapy for men with BPH.

Investigating microRNA Profiles in Prostate Cancer Bone Metastases and Functional Effects of microRNA-23c and microRNA-4328.

MicroRNAs (miRNAs) are aberrantly expressed in prostate cancer (PC), but comprehensive knowledge about their levels and function in metastatic PC is lacking. Here, we explored the differential expression of miRNA profiles during PC progression to bone metastasis, and further focused on the downregulation of miRNA-23c and -4328 and their impact on PC growth in experimental models. Using microarray screening, the levels of 1510 miRNAs were compared between bone metastases (n = 14), localized PC (n = 7) and benign prostate tissue (n = 7). Differentially expressed miRNAs (n = 4 increased and n = 75 decreased, p < 0.05) were identified, of which miRNA-1, -23c, -143-3p, -143-5p, -145-3p, -205-5p, -221-3p, -222-3p and -4328 showed consistent downregulation during disease progression (benign > localized PC > bone metastases). The downregulation of miRNA-23c and -4328 was confirmed by reverse transcription and quantitative polymerase chain reaction analysis of 67 metastasis, 12 localized PC and 12 benign prostate tissue samples. The stable overexpression of miRNA-23c and -4328 in the 22Rv1 and PC-3 cell lines resulted in reduced PC cell growth in vitro, and in the secretion of high levels of miRNA-23c (but not -4328) in extracellular vesicles. However, no tumor suppressive effects were observed from miRNA-23c overexpression in PC-3 cells subcutaneously grown in mice. In conclusion, bone metastases display a profound reduction of miRNA levels compared to localized PC and benign disease. The downregulation of those miRNAs, including miRNA-23c and -4328, may lead to a loss of tumor suppressive effects and provide biomarker and therapeutic possibilities that deserve to be further explored.

Risk factors for bladder neck contracture after transurethral resection of the prostate.

Transurethral resection of the prostate (TURP) is the most frequently used treatment of benign prostate hyperplasia with a prostate volume of <80 mL. A long-term complication is bladder neck contracture (BNC). The aim of the present study was to identify the risk factors for BNC formation after TURP. We conducted a retrospective analysis of all TURP primary procedures which were performed at one academic institution between 2013 and 2018. All patients were analyzed and compared with regard to postoperative formation of a BNC requiring further therapy. Uni- and multivariable logistic regression analyses (MVAs) were performed to identify possible risk factors for BNC development. We included 1368 patients in this analysis. Out of these, 88 patients (6.4%) developed BNC requiring further surgical therapy. The following factors showed a statistically significant association with BNC development: smaller preoperative prostate volume (p = 0.001), lower resected prostate weight (p = 0.004), lower preoperative levels of prostate-specific antigen (PSA, p < 0.001), shorter duration of the surgery (p = 0.027), secondary transurethral intervention (due to urinary retention or gross hematuria) during inpatient stay (p = 0.018), positive (≥100 CFU/mL) preoperative urine culture (p = 0.010), and urethral stricture (US) formation requiring direct visual internal urethrotomy (DVIU) postoperatively after TURP (p < 0.001), in particular membranous (p = 0.046) and bulbar (p < 0.001) strictures. Preoperative antibiotic treatment showed a protective effect (p = 0.042). Histopathological findings of prostate cancer (PCA) in the resected prostate tissue were more frequent among patients who did not develop BNC (p = 0.049). On MVA, smaller preoperative prostate volume (p = 0.046), positive preoperative urine culture (p = 0.021), and US requiring DVIU after TURP (p < 0.001) were identified as independent predictors for BNC development. BNC is a relevant long-term complication after TURP. In particular, patients with a smaller prostate should be thoroughly informed about this complication.

The Relationship Between Metabolic Syndrome and Benign Prostate Enlargement: A Case-Control Study in a Peruvian Military Hospital.

This study aimed to determine how metabolic syndrome is related to benign prostatic enlargement in males under 60 years old in a national military hospital in Peru. We conducted a retrospective, quantitative, case-control study. By simple random sampling, 87 cases with benign prostatic enlargement and 174 controls were included, with a statistical power of 80%. The benign prostatic enlargement was evaluated by clinical picture and ultrasound and the metabolic syndrome was evaluated according to the Adult Treatment Panel III criteria. The statistical analysis was performed using the STATAv14 program, the chi-square statistical test was used and odds ratio was obtained, at a significance level of 5%. The mean age of the cases and controls was 55 (51-58) and 52 (46-57), respectively. By multivariate analysis, the factors related to benign prostatic enlargement were the presence of benign prostatic enlargement (adjusted odds ratio: 2.71, 95% CI: 1.27-5.80; P=.010), waist circumference ≥102 cm (adjusted odds ratio: 6.51, 95% CI: 3.09-13.71; P < .001), elevated fasting glucose (adjusted odds ratio: 1.38, 95% CI: 0.65-2.91; P=.399), high triglycerides (adjusted odds ratio: 5.29, 95%: CI 2.40-11.64; P < .001), and arterial hypertension (adjusted odds ratio: 4.67, 95% CI 2.19-9.95; P < .001). Elevated high-density lipoprotein cholesterol was a protective factor (adjusted odds ratio: 0.09, 95% CI: 0.04-0.20; P < .001). The present study showed that metabolic syndrome and its components (waist circumference, hypertension, triglycerides, and high-density lipoprotein cholesterol) are factors related to benign prostatic enlargement in patients under 60 years old in a military hospital in Peruvian population. Waist circumference as an indicator of overweight/obesity is a practical anthropometric marker of interest in public health.

Cell-by-cell quantification of the androgen receptor in benign and malignant prostate leads to a better understanding of changes linked to cancer initiation and progression.

The androgen receptor (AR) plays a crucial role in the development and homeostasis of the prostate and is a key therapeutic target in prostate cancer (PCa). The gold standard therapy for advanced PCa is androgen deprivation therapy (ADT), which targets androgen production and AR signaling. However, resistance to ADT develops via AR-dependent and AR-independent mechanisms. As reports on AR expression patterns in PCa have been conflicting, we performed cell-by-cell AR quantification by immunohistochemistry in the benign and malignant prostate to monitor changes with disease development, progression, and hormonal treatment. Prostates from radical prostatectomy (RP) cases, both hormone-naïve and hormone-treated, prostate tissues from patients on palliative ADT, and bone metastases were included. In the normal prostate, AR is expressed in >99% of luminal cells, 51% of basal cells, and 61% of fibroblasts. An increase in the percentage of AR negative (%AR-) cancer cells along with a gradual loss of fibroblastic AR were observed with increasing Gleason grade and hormonal treatment. This was accompanied by a parallel increase in staining intensity of AR positive (AR+) cells under ADT. Staining AR with N- and C-terminal antibodies yielded similar results. The combination of %AR- cancer cells, %AR- fibroblasts, and AR intensity score led to the definition of an AR index, which was predictive of biochemical recurrence in the RP cohort and further stratified patients of intermediate risk. Lastly, androgen receptor variant 7 (ARV7)+ cells and AR- cells expressing neuroendocrine and stem markers were interspersed among a majority of AR+ cells in ADT cases. Altogether, the comprehensive quantification of AR expression in the prostate reveals concomitant changes in tumor cell subtypes and fibroblasts, emphasizing the significance of AR- cells with disease progression and palliative ADT.

Genetically predicted alterations in thyroid function are associated with the risk of benign prostatic disease.

Benign prostatic diseases (BPDs), such as benign prostate hyperplasia (BPH) and prostatitis, harm the quality of life of affected patients. However, observational studies exploring the association between thyroid function and BPDs have hitherto yielded inconsistent results. In this study, we explored whether there is a causal genetic association between them using Mendelian randomization (MR) analysis. We used publicly available summary statistics from the Thyroidomics Consortium and 23andMe on thyrotropin (TSH; 54,288 participants), thyroxine [free tetraiodothyronine (FT4); 49,269 participants], subclinical hypothyroidism (3,440 cases and 49,983 controls), overt hypothyroidism (8,000 cases and 117,000 controls), and subclinical hyperthyroidism (1,840 cases and 49,983 controls) to screen for instrumental variables of thyroid function. Results for BPD such as prostatic hyperplasia (13,118 cases and 72,799 controls) and prostatitis (1,859 cases and 72,799 controls) were obtained from the FinnGen study. The causal relationship between thyroid function and BPD was primarily assessed using MR with an inverse variance weighted approach. In addition, sensitivity analyses were performed to test the robustness of the results. We found that TSH [OR (95% CI) = 0.912(0.845-0.984), p =1.8 x 10-2], subclinical hypothyroidism [OR (95% CI) = 0.864(0.810-0.922), p =1.04 x 10-5], and overt hypothyroidism [OR (95% CI) = 0.885 (0.831-0. 944), p =2 x 10-4] had a significant effect on genetic susceptibility to BPH, unlike hyperthyroidism [OR (95% CI) = 1.049(0.990-1.111), p =1.05 x 10-1] and FT4 [OR (95% CI) = 0.979(0.857-1.119), p = 7.59 x 10-1] had no effect. We also found that TSH [OR (95% CI) =0.823(0.700-0.967), p = 1.8 x 10-2] and overt hypothyroidism [OR (95% CI) = 0.853(0.730-0.997), p = 4.6 x 10-2] significantly influenced the prostatitis, whereas FT4 levels [OR (95% CI) = 1.141(0.901-1.444), p = 2.75 x 10-1], subclinical hypothyroidism [OR (95% CI) =0. 897(0.784- 1.026), p = 1.12 x 10-1], and hyperthyroidism [OR (95% CI) = 1.069(0.947-1.206), p = 2.79 x 10-1] did not have a significant effect. Overall, our study results suggest that hypothyroidism and TSH levels influence the risk of genetically predicted BPH and prostatitis, providing new insights into the causal relationship between thyroid function and BPD.

A prognostic signature consisting of N6-methyladenosine modified mRNAs demonstrates clinical potential in prediction of biochemical recurrence and guidance on precision therapy in prostate cancer

Novel biomarkers are urgently needed to improve the prediction of clinical outcomes and guide personalized treatment for prostate cancer (PCa) patients. However, the role of N6-methyladenosine (m6A) modifications in PCa initiation and progression remains largely elusive. In our study, we collected benign Prostate Hyperplasia (BPH), localized PCa, and metastatic PCa samples from patients and performed methylated RNA immunoprecipitation sequencing (MeRIP-Seq) to map m6A-methylated mRNAs. Furthermore, we developed a prognostic signature based on 239 differentially methylated RNAs and the TCGA-PRAD dataset, which can be used to calculate an m6A-modified mRNA (MMM) score for a PCa patient, validated by independent multi-center cohorts. Our findings revealed that differential m6A modifications were positively correlated with altered expressions of mapped m6A-modified mRNAs. Higher MMM scores were associated with shorter times to biochemical recurrence (BCR) in PCa patients, and the MMM scoring system outperformed three well-established signatures in nine independent validation cohorts, as demonstrated by Kaplan-Meier survival analysis, C-index and ROC. Patients who did not respond to androgen receptor signaling inhibitor (ARSI) therapy and immunotherapy were found to have high MMM scores. Two hub genes, TLE1 and PFKL, were confirmed to have m6A sites through MeRIP-qPCR, and their knockdown promoted PCa cell invasion. Bioinformatics analysis of single-cell databases identified cell types with high transcript abundance levels of these two genes. In summary, our study is the first to perform transcriptome-wide m6A mapping in prostate tissues. The translational potential of a prognostic signature, comprising m6A-methylated mRNAs, in predicting clinical outcomes and therapy responses for PCa patients, is demonstrated.

Evaluation of Matrix Metalloproteases by Artificial Intelligence Techniques in Negative Biopsies as New Diagnostic Strategy in Prostate Cancer.

Usually, after an abnormal level of serum prostate-specific antigen (PSA) or digital rectal exam, men undergo a prostate needle biopsy. However, the traditional sextant technique misses 15-46% of cancers. At present, there are problems regarding disease diagnosis/prognosis, especially in patients' classification, because the information to be handled is complex and challenging to process. Matrix metalloproteases (MMPs) have high expression by prostate cancer (PCa) compared with benign prostate tissues. To assess the possible contribution to the diagnosis of PCa, we evaluated the expression of several MMPs in prostate tissues before and after PCa diagnosis using machine learning, classifiers, and supervised algorithms. A retrospective study was conducted on 29 patients diagnosed with PCa with previous benign needle biopsies, 45 patients with benign prostatic hyperplasia (BHP), and 18 patients with high-grade prostatic intraepithelial neoplasia (HGPIN). An immunohistochemical study was performed on tissue samples from tumor and non-tumor areas using specific antibodies against MMP -2, 9, 11, and 13, and the tissue inhibitor of MMPs -3 (TIMP-3), and the protein expression by different cell types was analyzed to which several automatic learning techniques have been applied. Compared with BHP or HGPIN specimens, epithelial cells (ECs) and fibroblasts from benign prostate biopsies before the diagnosis of PCa showed a significantly higher expression of MMPs and TIMP-3. Machine learning techniques provide a differentiable classification between these patients, with greater than 95% accuracy, considering ECs, being slightly lower when considering fibroblasts. In addition, evolutionary changes were found in paired tissues from benign biopsy to prostatectomy specimens in the same patient. Thus, ECs from the tumor zone from prostatectomy showed higher expressions of MMPs and TIMP-3 compared to ECs of the corresponding zone from the benign biopsy. Similar differences were found for expressions of MMP-9 and TIMP-3, between fibroblasts from these zones. The classifiers have determined that patients with benign prostate biopsies before the diagnosis of PCa showed a high MMPs/TIMP-3 expression by ECs, so in the zone without future cancer development as in the zone with future tumor, compared with biopsy samples from patients with BPH or HGPIN. Expression of MMP -2, 9, 11, and 13, and TIMP-3 phenotypically define ECs associated with future tumor development. Also, the results suggest that MMPs/TIMPs expression in biopsy tissues may reflect evolutionary changes from prostate benign tissues to PCa. Thus, these findings in combination with other parameters might contribute to improving the suspicion of PCa diagnosis.

Targeting SMYD2 inhibits prostate cancer cell growth by regulating c-Myc signaling.

SMYD2 is a lysine histone methyl transferase involved in various cancers epigenetically via methylating histone H3K4, and H3K36. c-Myc is one of the major drivers ofprostate cancer (PCa) initiation and progression. The roles of SMYD2 in PCa and the regulators of c-Myc activity in PCa are still under-researched.SMYD2 expression and survival outcomes in PCa cohorts were analyzed by bioinformatics analysis. SMYD2 protein levels were detected in PCa tissues by immunohistochemistry. SMYD2 knockdown cells were established to identify the effects of SMYD2 on cell growth in vitro and in vivo. GSEA and RNA sequencing were adopted to reconnoiter the signaling regulated by SMYD2 in PCa. The relationship between SMYD2 and c-Myc was examined by western blot analysis, qPCR, and immunohistochemistry. SMYD2 specific inhibitor-AZ505 was used to pharmacologically inhibit SMYD2 function in vitro and in vivo.SMYD2 expression increased in PCa tissues compared with benign prostate tissues and higher SMYD2 expression was associated with a higher risk of biochemical relapse after radical prostatectomy. SMYD2 knockdown inhibited the growth of PCa cells both in vitro and in vivo. Furthermore, high SMYD2 levels conduced to activated c-Myc signaling in PCa cells. Importantly, the pharmacological intervention of SMYD2 by AZ505 significantly repressed PCa cell growth both in vitro and in vivo. Our findings indicate that SMYD2 inhibition restrains PCa cell proliferation by regulating c-Myc signaling and provide evidence for the potential practice of SMYD2 targeting in the treatment of PCa.

Abstract 4745: Prostatic fluid-based cfDNA methylation profiling distinguish benign hyperplasia from prostate cancer

Abstract Introduction: A challenge in the early diagnosis of prostate cancer (PC) is to distinguish between malignant tumor and benign prostatic hyperplasia (BPH) because of the similar symptoms. Methylation profiles provide a way for discriminating BPH from PC, which could prevent overdiagnosis and overtreatment. Applying methylation biomarkers to liquid biopsy specimens provides higher sensitivity to detect tumors. Methods: Prostatic fluid samples were prospectively collected from 40 patients following digital rectal exam and isolated cell-free DNA to perform whole genome methylation sequencing using PredicineEPIC platform. Combing differentially methylated regions (DMR) with methylation signature identified from massive public data, a panel of methylation patterns was developed. The difference of methylation signal in profiled region were implemented by machine learning to robustly distinguish BPH from prostate cancer. Results: We identified 324 biomarkers from prostatic fluid samples and collected 46 methylation biomarkers reported from previous research. Regions with extreme difference between benign prostatic hyperplasia and prostate cancer were identified as candidate biomarkers. A diagnostic model trained on these 55 biomarkers could distinguish BPH from PC in 10-fold cross-validation (AUC = 0.92). And this model could be applied for discriminating normal tissue from prostate tumor tissue with highly performance (AUC &amp;gt; 0.95). Using this set of regions, we developed an NGS-based panel for detecting methylation alteration of cfDNA from prostate liquid, which inherited the ability of differentiation. Conclusion: These methylation patterns could be developed as the novel biomarkers to support the development of prostatic fluid-based non-invasive diagnostic test and reduce unnecessary biopsies. This panel could be applied for differential diagnosis of benign hyperplasia and prostate cancer. Citation Format: Hang Dong, Haoran Tang, Yue Zhang, Shidong Jia. Prostatic fluid-based cfDNA methylation profiling distinguish benign hyperplasia from prostate cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4745.

Abstract 2196: Serum Filamin A is a prognostic biomarker for screening benign prostatic hyperplasia vs prostate cancer in Caucasian and African American men

Abstract Prostate cancer represents a significant health risk to aging men, in which diagnostic challenges pose major issues with identification of indolent and or aggressive cancers. Prostate cancer screening is driven by prostate specific antigen (PSA), however, PSA’s screening utility is diminished in men with benign prostatic hyperplasia (BPH) due to an enlarged prostate and elevated PSA, triggering unnecessary biopsies in men with this condition. Thus, there is a critical unmet medical need to identify a marker for men with these conditions. Moreover, it is crucial to test any novel biomarker in populations of different ancestry (especially African American men who have higher prostate cancer risk) to ensure results apply regardless of race. To this extent, we recently identified a cleaved fragment of Filamin A (FLNA) protein (as measured by IP-MRM mass spectrometry assessment) as a novel biomarker for stratifying BPH from prostate cancer and subsequently evaluated its utility in Caucasian and African American patients. The men had a negative digital rectal examination (DRE), PSA between 4-10 ng/ml and all underwent prostate biopsy. In African American (AA) men, FLNA expression exhibited diagnostic utility for stratifying BPH from all prostate cancer patients (0.72 AUC, 0.82 PPV, 0.76 NPV, 14.2 OR, P Value - 1.5 e-08 in 48 BPH and 139 PCa AA men) and outperformed PSA (0.54 AUC, 0.75 PPV, 0.28 NPV, 1.1 OR, P Value 0.78). In addition, FLNA could stratify BPH from Gleason 7-10 prostate cancers (0.71 AUC, 0.66 PPV, 0.86 NPV, 12.2 OR, P Value 1.7 e-05 in 48 BPH and 60 PCa AA men) and again outperformed PSA (0.5 AUC, 0.58 PPV, 0.62 NPV, 2.18 OR, P Value 0.24). In Caucasian American (CA) men, FLNA expression exhibited diagnostic utility for stratifying BPH from all prostate cancer patients (0.68 AUC, 0.68 PPV, 0.73 NPV, 5.6 OR, P Value 1.6 e-12 in 191 BPH and 281 PCa CA men) and outperformed PSA (0.56 AUC, 0.58 PPV, 0.28 NPV, 0.55 OR, P Value 0.13). FLNA could also stratify BPH from Gleason 7-10 prostate cancers (0.74 AUC, 0.46 PPV, 0.96 NPV, 19.4 OR, P Value 1.6 e-12 in 119 BPH and 109 PCa CA men) and again outperformed PSA (0.46 AUC, 0.35 PPV, 0.38 NPV, 0.32 OR, P Value 0.03). Previously we showed FLNA along with prostate volume had superior performance over PSA in identifying patients with prostate cancer (Kiebish et al Scientific Reports, 2021 Jul 23, 11(1)). Herein, we refined our analysis and establish FLNA alone as a serum biomarker for stratifying men with BPH vs prostate cancer as well as those with high Gleason prostate cancers compared to the current diagnostic paradigm of using PSA. This simplified approach demonstrates clinical actionability as a test with utility in AA and CA men and represents a significant opportunity to decrease the need for unnecessary biopsies for prostate cancer diagnosis. A clinical test is being developed to address this important medical need. Citation Format: Nischal Mahaveer Chand, Poornima K. Tekumalla, Albert Dobi, Amina Ali, Gregory M. Miller, Juan J. Aristizabal-Henao, Elder Granger, Stephen J. Freedland, Shiv Srivastava, Jose Arturo Rodriguez Rivera, Arturo Mendoza, David G. McLeod, Niven R. Narain, Michael A. Kiebish. Serum Filamin A is a prognostic biomarker for screening benign prostatic hyperplasia vs prostate cancer in Caucasian and African American men [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2196.

Abstract 4868: Targeting of ERG positive prostate cancers with ERGi-USU-6 salt derivative

Abstract Introduction: Prostate cancer (PCa) is the second leading cause of cancer deaths among men in the United States. Approximately 50% of patients with PCa harbor an oncogenic TMPRRS2- ERG gene fusion in their primary tumor and 35% of patients with metastatic castration resistant prostate cancers have the gene fusion. We have identified a potent small molecule inhibitor, ERGi-USU-6 salt derivative 7b, that selectively inhibits the growth of ERG positive tumor. This small molecule inhibitor is also effective in inhibiting the growth of benign and cancerous mouse prostate organoids expressing TMPRSS2-ERG (ERG positive organoids). To gain insights into the cancer-selective properties of ERGi-USU-6 salt 7b we evaluated pathways associated in ERGi-USU induced inhibition. Methods: The biological activities of salt derivative 7b, were assessed in hormone- refractory metastatic tumor derived ERG positive prostate cancer cell line, VCaP along with organoids. We monitored the pathways associated in the mechanism of drug action through, cell cycle-regulator proteins by immunoblot assays, cell cycle and ferroptosis related analyses in response to 7b treatment. We also monitored the levels of the RIOK2 kinase, previously shown to bind the parental ERGi-USU compound. The normal primary endothelium derived HUVEC cells were used as normal control due to the normal endogenous expression of ERG in endothelial cells including HUVEC. Results: Cell growth and immunoblot analysis indicated the inhibition of ERG positive prostate organoid upon ERGi-USU treatments resulting in the downregulation of ERG and RIOK2 protein levels. The cell cycle analyses, pathway mapping by protein assessment and ferroptotic assays suggests that salt derivative 7b treatment inhibits the ERG positive prostate cancer through ferroptosis along with RIOK2 inhibition. Conclusions: Our results showed that the ferroptosis inducer ATF3 gene is involved in the cancer-selective activity of ERGi-USU-6 salt derivative 7b. Further, based on our observations we hypothesize that ferroptosis, the iron-dependent form of programmed cell-death, may be the mechanism of cancer selective activity of salt derivative 7b. Disclaimer: The contents of this publication are the sole responsibility of the author(s) and do not necessarily reflect the views, opinions or policies of Uniformed Services University of the Health Sciences (USUHS), The Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., the Department of Defense (DoD), the Departments of the Army, Navy, or Air Force. Mention of trade names, commercial products, or organizations does not imply endorsement by the U.S. Government. Citation Format: Binil Eldhose, Katherine Beck, Cyrus Eghtedari, Gartrell C. Bowling, Mallesh Pandrala, Sanjay V. Malhotra, Xiaofeng A. Su, Albert Dobi. Targeting of ERG positive prostate cancers with ERGi-USU-6 salt derivative. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4868.

Abstract 5292: Characterization of androgen receptor properties and interactome in mediating transcriptional condensates in prostate cancer

Abstract In The United States of America 1 in 8 men will develop prostate cancer (PCa) in their lifetime. The assembly of phase-separated condensates containing players of the transcriptional machinery such as transcription factors, coactivators (e.g. MED1), and RNA polymerase II enhances the transcription of key oncogenes in various cancer including PCa. Many proteins undergoing phase-separation possess intrinsically disordered regions (IDR) that mediate multivalent intra- and inter-molecular interactions, essential for condensate formation. We recently demonstrated that full-length AR, the main oncogenic driver in PCa, is more prone to form nuclear transcriptional condensates upon androgen stimulation in PCa models than in benign epithelial prostate models. In this study, we investigated the effects of mutating or truncating key residues or regions involved in AR transcriptional activity on its ability to form nuclear condensates in LNCaP cells as visualized by confocal microscopy. We also used various bioinformatics tools to predict additional AR residues and sequences with high propensity for phase-separation. We thus made various truncations and mutations and evaluated their ability to form condensates in LNCaP cells. We also tested the effect MED1 phosphorylation and treatment with various inhibitors targeting different domains of the AR on the ability of full-length protein to form droplets in vitro. We hope by better understanding what drive AR condensates formation in PCa, to elucidate this new mechanism of transcriptional regulation and to identify new therapeutic avenues for patients with advanced forms of the disease. Citation Format: Nicholas C. Pinette, Shabnam Massah, Sofia Kochkina, Fan Zhang, Maitree Biswas, Joseph Lee, Jörg Gsponer, Nada Lallous. Characterization of androgen receptor properties and interactome in mediating transcriptional condensates in prostate cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5292.

Abstract 5551: Cancer associated PSA-glycoforms as prostate cancer markers

Abstract Changes in protein glycosylation have been observed in several cancers, where they affect cellular growth behavior, invasiveness and acquisition of metastatic potential. Such changes are not random and have been found to associate with cancer aggressiveness. Thus, detection of protein glycosylation may offer novel opportunities for cancer biomarker development. Since prostate-specific antigen (PSA) is glycosylated, assessing different PSA-glycoforms may provide valuable diagnostic and prognostic information. We recently established a novel in situ proximity ligation-based method for the detection of different protein-glycoforms in tissue sections. This method utilizes protein-specific antibody and glycan-binding lectins. We have optimized this method for the detection of different PSA-glycoforms in formalin-fixed paraffin-embedded prostatic tumor samples. We found that PSA is, indeed, differently glycosylated in prostate cancer (n=23) compared to benign prostate tissue (n=4). Two of the 25 studied lectins showed prominent staining only in cancer tissues (p&amp;lt;0.0001 for both), although the total PSA staining was stronger in benign tissues (p=0.006). We are currently validating these results in large, well-curated clinical cohorts, with intention to test whether PSA-glycoforms in tissue offer improved detection of clinically significant prostate cancer. These studies facilitate development of tools for identification of cancer-specific PSA-isoforms in serum samples. Detection of such isoforms is likely to provide independent diagnostic and prognostic information. Citation Format: Hannu Koistinen, Ruusu-Maaria Kovanen, Timo-Pekka Lehto, Antti Rannikko, Tuomas Mirtti. Cancer associated PSA-glycoforms as prostate cancer markers. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5551.

Ejaculation-sparing enucleation of benign prostate hyperplasia: is it almost feasible?

Introduction. Patients often request maintenance of antegrade ejaculation, and try to find the clinic, surgeon, and treatment to store it. Despite an established technique, the long-term results of ejaculation-sparing operations at a large prostate volume remain unstable. This is not allowed to give patients a guarantee in maintaining this component in their sexual life.Objective. To evaluate the frequency of ejaculation maintenance after laser ejaculatory-sparing prostate enucleation.Materials &amp; methods. Since 2017 ejaculation-sparing laser enucleations have been performed in prostate volume (V pr &gt; 80 cc) in 84 patients. Fifty-four patients (64%) had three lobes benign prostate hyperplasia (BPH) and two lobes — 30 patients (36%). Preoperative voiding parameters were IPSS score — 21.0 ± 2.7, QoL — 4.8 ± 0.6, residual urine volume (V res) — 139 ± 43 mL, Q max — 8.1 ± 2.0 mL. We estimate the result as positive if antegrade ejaculation remained after intervention. If ejaculation volume decreased, then as partially positive and negative — ejaculation was absent postoperatively. We conducted questionnaire and follow-up examination 3 – 6 months after intervention.Results. Thulium-fiber laser enucleations have been performed in all cases without any serious complications. Followup 3 – 6 months after operation showed V pr — 29 ± 4 cc, V res — 19 ± 17 mL, Q max — 19.1 ± 3.1 mL/sec. IPSS score — 8.1 ± 1.9 and QoL — 2.6 ± 0.7. There are the following technical remarks: 1) tissue-sparing in the verumontanum; 2) no or minor mechanical tissue tension during enucleation; 3) avoid additional resection in the bladder neck; 4) refuse total coagulation; 5) no urethral catheter tension after operation; 6) catheter balloon inflow only in the bladder, not in the fossa. Antegrade ejaculation has stored at 17 (20%) patients, partial ejaculation — at 34 (40%) patients, so overall efficacy is 60%. The presence of a median lobe is shown to be negative prognostic factor due to the absence of full antegrade ejaculation in all cases. A partial ejaculation was achieved at 21 patients. Based on the obtained results we have optimized the three-lobe prostate enucleation technique. Four from our initial 7 patients had full storage of ejaculation and 3 — partial.Conclusion. The possibility of BPH laser surgery to store ejaculation function at the request of the patients is modest. The presence of the median lobe makes the full ejaculation-sparing impossible. In cases of two lobes BPH full storage of the ejaculation was at 57% and partial at 43%, respectively. The following evaluation of the described technique is mandatory.

Abstract 520: Investigation the function and application of spermine in castration-resistant prostate cancer

Abstract Introduction &amp; Objectives: Spermine (Spm) is a typical metabolite that is highly concentrated in healthy prostate epithelium maintaining cell growth and functions. Our previous studies found that Spm was significantly decreased in PCa tissue than in benign prostate tissue, as was the urinary Spm in PCa. This study aimed to reveal the specific role of Spm in PCa. Materials &amp; Methods: In this study, Spm was resupplied to castration-resistant prostate cancer (CRPC) cells (PC3 and DU145). Following Spm treatment, MTT (methyl thiazolyl tetrazolium) and EdU (5-ethynyl-2’-deoxyuridine) assay were used to determine the cell proliferation of CRPC cell lines. Cellular apoptosis, cell cycle distribution and mitochondrial membrane potential were analyzed by flow cytometry. Cellular reactive Oxygen species (ROS) level was measured by fluorescence microscope. mRNA and ATAC sequencing in spermine-treated CRPC cells were performed to reveal the related mechanism for Spm effect. Results: In the in vitro studies, Spm was found to inhibit the cell viability of CRPC cells. Transwell assays demonstrated that Spm impaired migration and invasion ability. And Spm blocked tumor progression by inducing G2/M arrest and cellular apoptosis in CRPC cells dose-dependently. mRNA seq revealed that a series of genes were altered after Spm treatment and indicated the potential functional pathway of Spm might be mitophagy. We integrated mRNA seq data with ATAC seq profiles and accessed those genes regulated by polyamine metabolism and Spm-induced mitophagy. Results showed the sequencing tracks for the HMOX1 locus contain higher ATAC-seq peaks at the promoter under Spm treatment. Spm could affect oxidative phosphorylation which will lead to mitophagy and mitochondrial metabolism by modulating the transcriptional accessibility of genes. After Spm treatment, cellular ROS was upregulated while the mitochondrial membrane potential collapsed. Subsequently, western blot results showed that PINK1-mediated mitophagy related-proteins were significantly regulated. Conclusion: Our results indicated that Spm treatment led to mitochondrial dysfunction and subsequently degraded through mitophagy in CRPC cells. Our findings could provide a theoretical basis and have preliminary clinical implications in developing Spm as a promising agent against CRPC by inducing cellular mitophagy in CRPC cells. Citation Format: Jingkai Sun, Peter Ka-Fung Chiu, Tingting Xie, Jeremy Yuen-Chun Teoh, Chi-Fai Ng. Investigation the function and application of spermine in castration-resistant prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 520.

Abstract 2453: Olfactomedin 4 downregulation enhances MYC-signaling in human benign prostate cells and the Olfm4 mouse models

Abstract Prostate cancer is the most diagnosed solid tumor and the second leading cause of cancer related death in American men. We have reported previously that OLFM4 gene expression was reduced or lost during prostate cancer progression due to frequent genetic deletion and hypermethylation of the OLFM4 gene promoter region. To investigate OLFM4 gene biological functions and molecular mechanisms underlying prostate cancer progression, we have established OLFM4 knock out RWPE1 cells and Olfm4 knock out mouse model. Functionally, OLFM4-knockout RWPE1 cells exhibited enhanced proliferation of the stem/progenitor-like cell population and abnormal differentiation. Bulk-cell RNA sequencing analysis pinpointed that MYC expression was enhanced in the OLFM4-knockout RWPE1 cells compared with OLFM4-wild cells. Molecular and signaling pathway studies revealed an increase in the WNT/APC/MYC signaling pathway gene signature and MYC target genes that regulate multiple biological processes in OLFM4-knockout RWPE1 cells and Olfm4-knock out mouse models. To test the function of the MYC gene in RWPE1 cells, we used (+)-JQ1, an MYC inhibitor, in both 2D and 3D culture models and found that (+)-JQ1 substantially inhibited the proliferation of OLFM4-knockout GFP reporter RWPE1 cells compared with OLFM4-W RWPE1 cells in both types of cultures. We further studied MYC inhibitors using organoid culture for Olfm4-/-/TRAMP mouse prostate tumors. These results suggest that OLFM4-negative prostate cancer is more sensitive to MYC inhibitor’s therapeutic approaches. Citation Format: Hongzhen Li, Wenli Liu, Jianqiong Zhu, Kay Chin, Griffin P. Rodgers. Olfactomedin 4 downregulation enhances MYC-signaling in human benign prostate cells and the Olfm4 mouse models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2453.

Abstract 3771: PiR-hsa-4447944 promotes the progression of prostate cancer through NEFH and induces androgen-independent growth

Abstract Introduction and Objective: Non-coding RNAs (ncRNAs) represent a diverse family of regulatory transcripts that drive tumorigenesis of prostate cancer (PCa) and various other types of cancers by their hyperactivity or diminished function. PIWI-interacting RNAs (piRNAs) are a class of small ncRNAs mechanistically similar to but much less studied than microRNAs. As the most diversified small ncRNAs, piRNAs are widely involved in the pathogenesis of different types of cancers. Our study aimed to investigate the role of piRNAs in the development of castration-resistant prostate cancer (CRPC). Methods: Small RNA sequencing was used to explore the CRPC associated piRNAs on a basis of 10 benign prostate tissues, 10 paired hormone-sensitive PCa tissues and CRPC tissues from the same patients. The top 12 upregulated piRNAs were validated through qRT-PCR in a bigger cohort. piR-hsa-4447944 was selected and investigated its biological function by gain of function in vitro. To study its loss of function role, an androgen-independent PCa cell (LNCaP-AI) was established. For mechanistic investigation, high-throughput RNA sequencing was conducted to identify dysregulated genes in PCa cells with control or piR-hsa-4447944 overexpression. Then, potential downstream targets were further predicted by miRanda program and validated by qRT-PCR, western-blot and dual-luciferase reporter assay. Results: piR-hsa-4447944 demonstrated a relatively higher expression level in both CRPC tissues and CRPC cell lines when compared with PCa tissues and cells. Functionally, overexpression of piR-hsa-4447944 could confer resistance to androgen deprivation and anti-androgen in PCa cells in vitro, whereas anti-sense RNA against the piR-hsa-4447944 could potentiate the sensitivity to androgen deprivation in LNCaP-AI cell. In addition, piR-hsa-4447944 showed the ability in promoting migration and invasion of PCa cells. Mechanistically, mRNA sequencing results identified a panel of downstream target-genes. Among them, neurofilament heavy chain (NEFH) was predicted and validated to have a direct binding relationship with piR-hsa-4447944, and was successfully verified to be negatively regulated by piR-hsa-4447944 in PCa cells. Conclusions: Our study showed, for the first time, that piR-hsa-4447944 can contribute to androgen insensitivity in CRPC. This sheds a new insight into the regulation of CRPC by a small ncRNA piRNA, which may form the basis of a novel target in the treatment of CRPC. Citation Format: Qiang Peng, Jingkai Sun, Peter Ka-Fung Chiu, Tingting Xie, Jeremy Yuen-Chun Teoh, Chi-Fai Ng. PiR-hsa-4447944 promotes the progression of prostate cancer through NEFH and induces androgen-independent growth. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3771.

MP76-01 MID-TERM EVALUATION OF IMPACT ON STORAGE AND OBSTRUCTIVE SYMPTOMS WITH REZŪM: OUTCOMES OF A LARGE, MULTI-CENTER, PROSPECTIVE REGISTRY OF REZŪM WATER VAPOR THERAPY BENIGN PROSTATIC ENLARGEMENT

You have accessJournal of UrologyCME1 Apr 2023MP76-01 MID-TERM EVALUATION OF IMPACT ON STORAGE AND OBSTRUCTIVE SYMPTOMS WITH REZŪM: OUTCOMES OF A LARGE, MULTI-CENTER, PROSPECTIVE REGISTRY OF REZŪM WATER VAPOR THERAPY BENIGN PROSTATIC ENLARGEMENT Kevin C. Zorn, Naeem Bhojani, Bilal Chughtai, and Dean Elterman Kevin C. ZornKevin C. Zorn More articles by this author , Naeem BhojaniNaeem Bhojani More articles by this author , Bilal ChughtaiBilal Chughtai More articles by this author , and Dean EltermanDean Elterman More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003350.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Rezūm convective water vapor ablation is a minimally invasive procedure to treat benign prostatic tissue. Herein, we report a prospective, multi-center results of the largest cohort of prostates treated with Rezum in Canada to address mid-term outcomes on IPSS storage and obstructive voiding symptoms. METHODS: A prospective registry was established for Rezūm therapy in Canada (2019) at two high-volume centers. All patients had baseline medical and BPH history documented along with uroflowmetry (Qmax and PVR), and validated questionnaires (IPSS, IPSS QoL, BPHII, IIEF-15, MSHQ-EjD function and bother). Sub-category IPSS-V and IPSS-S were evaluated for the entire cohort, men in retention and those with prostate volume >80 cc vs <80 cc and median lobe vs non-median lobe. IPSS-S was calculated by summing question 1 (incomplete emptying), question 3 (intermittency), question 5 (weak stream), and question 6 (straining to void). IPSS-V was calculated by summing question 2 (frequency), question 4 (urgency), and question 7 (nocturia). RESULTS: 345 men (mean age of 67.4, were treated with Rezūm from April 2019 to October 2022. The mean prostate volume was 72.3 cc (range 17-271) and 64% had a median lobe. 85 patients had prior episode of urinary. Mean number of injections was 10.1(range 2-28). Mean duration of procedure was 3.7 minutes (range 1-13). Mean duration of post-procedure catheterization was 9.8 days. IPSS-S reduced from a baseline score of 9.4 to 8.4 (10%), 6.1 (35%), 4.9 (48%), 4.8 (49%), 4.6 (51%), 4.3 (54%) at 1, 3, 6, 12, 24 and 36 months respectively. Comparatively, IPSS-V reduced from a baseline of 12.7 to 7.7 (39%), 4.1 (67%), 4.3 (66%), 4.3 (66%), 4.6 (64%), 3.7 (70%) at 1, 3, 6, 12, 24 and 35 months respectively. Outcomes were similar among subgroup analysis of men with baseline retention, men with prostate volume >80 cc or those with median lobe (all p>0.05). CONCLUSIONS: Rezum therapy is safe, effective, quick, out-patient, office-based procedure for prostate glands over a wide range of volumes with equally significant improvements in storage and voiding symptoms regardless of prostate volume, median lobe status and baseline urinary retention. Source of Funding: No © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e1090 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kevin C. Zorn More articles by this author Naeem Bhojani More articles by this author Bilal Chughtai More articles by this author Dean Elterman More articles by this author Expand All Advertisement PDF downloadLoading ...