Development Of Prostatic Adenocarcinoma Research Articles

USF1-mediated ALKBH5 stabilizes FLII mRNA in an m6A-YTHDF2-dependent manner to repress glycolytic activity in prostate adenocarcinoma.

Upstream-stimulating factor 1 (USF1) is a ubiquitously expressed transcription factor implicated in multiple cellular processes, including metabolism and proliferation. This study focused on the function of USF1 in glycolysis and the malignant development of prostate adenocarcinoma (PRAD). Bioinformatics predictions suggested that USF1 is poorly expressed in PRAD. The clinical PRAD samples revealed a low level of USF1, which was correlated with an unfavorable prognosis. Artificial upregulation of USF1 significantly repressed glycolytic activity in PRAD cells and reduced cell growth and metastasis in vitro and in vivo. Potential downstream genes of USF1 were probed by integrated bioinformatics analyses. The chromatin immunoprecipitation and luciferase assays indicated that USF1 bound to the α-ketoglutarate-dependent dioxygenase alkB homolog 5 (ALKBH5) promoter for transcription activation. Flightless I (FLII) was identified as the gene showing the highest degree of correlation with ALKBH5. As an m6A demethylase, ALKBH5 enhanced FLII mRNA stability by inducing m6A demethylation in an m6A-YTH N6-methyladenosine RNA-binding protein F2 (YTHDF2)-dependent manner. Either silencing of ALKBH5 or FLII blocked the role of USF1 in PARD cells and restored glycolysis, cell proliferation, and invasion. This study demonstrates that USF1 activates ALKBH5 to stabilize FLII mRNA in an m6A-YTHDF2-dependent manner, thereby repressing glycolysis processes and the progression of PRAD.

MRNA-Modified FUS/NRF2 Signalling Inhibits Ferroptosis and Promotes Prostate Cancer Growth.

Objective Regarding the imperfect mechanism of occurrence and development of prostate adenocarcinoma (PRAD), this study investigated mRNA-modified FUS/NRF2 signalling to inhibit ferroptosis and promote prostate adenocarcinoma growth. Methods Bioinformatics analysis was used to obtain the expression of FUS and its mRNA modification in PRAD. The expression of FUS in prostate cells (CRPC) and the level of m6A methylation modification, ferroptosis (P53 and GPX4), apoptosis (Caspase3), ferroptosis (P53 and GPX4), and apoptosis (Caspase3) in CRPC after ferroptosis inducer Erastin, ferroptosis inhibitor, and FUS knockdown were detected. Autophagy (LC3B), oxidative stress (GSH and ROS), and expression of NRF2/HO-1 pathway are indicators. Results FUS was highly expressed in PRAD and phenomenally reduced the survival rate of patients. After knocking down FUS, the level of m6A methylation was significantly reduced, and the expressions of ferroptosis markers P53 and GPX4 were phenomenally reduced, while the levels of apoptosis and autophagy markers Caspase3 and LC3B remained unchanged. Upregulated and NRF2/HO-1 pathway indicators were upregulated. It shows that m6A methylation modification is reduced when FUS is the low expression, inhibits the expression of P53 and GPX4, downregulates GSH, upregulates ROS, activates the NRF2/HO-1 pathway, and promotes ferroptosis to inhibit the occurrence of RPAD. Conclusions The increase of m6A methylation modification can increase the expression of FUS, thereby promoting the expression of P53 and GPX4, upregulating GSH, downregulating ROS, inhibiting the NRF2/HO-1 pathway, inhibiting ferroptosis, and promoting the growth of PRAD.

δ-Tocotrienol is the Most Potent Vitamin E Form in Inhibiting Prostate Cancer Cell Growth and Inhibits Prostate Carcinogenesis in Ptenp-/- Mice.

We demonstrated that δ-tocotrienol is the most active vitamin E form in inhibiting the growth of several prostate cancer cell lines. In transgenic Ptenp-/- mice, δ-tocotrienol inhibited the formation of prostate cancer. This result would encourage and help design clinical studies for the application of δ-tocotrienol for prostate cancer prevention.

High-fat diet-induced hyperinsulinemia promotes the development of prostate adenocarcinoma in prostate-specific Pten-/- mice.

Metabolic syndrome (MetS) and its four clinical entities, central obesity, insulin resistance, hypertension and dyslipidemia, are implicated in increasing the risk and mortality of cancer in several organs. However, it is unclear how they are associated with increased risk of prostate cancer. To elucidate the mechanistic link between MetS and prostate carcinogenesis, we characterized the development of MetS and prostate adenocarcinoma in prostate-specific Pten-/- (Ptenp-/-) mice fed a high-fat (HF) diet. We found that male Ptenp-/- mice on an HF diet gained excess body weight and elevated blood glucose, insulin and insulin-like growth factor 1 (IGF1) levels at 20 weeks of age and were obese at 40 weeks. Prostate adenocarcinoma multiplicity at 40 weeks was significantly higher in the mice on an HF diet, suggesting that the HF diet promotes the development of prostate adenocarcinoma. Increased cell proliferation and enhanced AKT activation were found in the prostates of mice on an HF diet. Further transcriptome study revealed that receptor tyrosine kinase regulation, which mediates insulin/IGF1 signaling, was one of the top enriched pathways by HF diet-induced transcriptome changes. Together, our results suggest that HF diet-induced hyperinsulinemia leads to increased activation of insulin/IGF1/AKT signaling in lesioned prostates, promoting the development of adenocarcinoma.

Novel immune-related signature for risk stratification and prognosis in prostatic adenocarcinoma.

A substantial proportion of prostatic adenocarcinoma (PRAD) patients experience biochemical failure (BCF) after radical prostatectomy (RP). The immune microenvironment plays a vital role in carcinogenesis and the development of PRAD. This study aimed to identify a novel immune‐related gene (IRG)‐based signature for risk stratification and prognosis of BCF in PRAD. Weighted gene coexpression network analysis was carried out to identify a BCF‐related module in a discovery cohort of patients who underwent RP at the Massachusetts General Hospital. The median follow‐up time was 70.32 months. Random forest and multivariate stepwise Cox regression analyses were used to identify an IRG‐based signature from the specific module. Risk plot analyses, Kaplan‐Meier curves, receiver operating characteristic curves, univariate and multivariate Cox regression analyses, stratified analysis, and Harrell’s concordance index were used to assess the prognostic value and predictive accuracy of the IRG‐based signature in the internal discovery cohort; The Cancer Genome Atlas database was used as a validation cohort. Tumor immune estimation resource database analysis and CIBERSORT algorithm were used to assess the immunophenotype of PRAD. A novel IRG‐based signature was identified from the specific module. Five IRGs (BUB1B, NDN, NID1, COL4A6, and FLRT2) were verified as components of the risk signature. The IRG‐based signature showed good prognostic value and predictive accuracy in both the discovery and validation cohorts. Infiltrations of various immune cells were significantly different between low‐risk and high‐risk groups in PRAD. We identified a novel IRG‐based signature that could function as an index for assessing tumor immune status and risk stratification in PRAD.

Abstract 1760: Role of Batf-dependant Th17 immune response in PTEN-null mouse model

Abstract Basic leucine zipper transcription factor ATF-like (BATF) belongs to the activator protein 1 (AP-1) family of transcription factors and has been shown to be predominantly expressed in cells of hematopoietic origin, especially in T cells. Mice deficient for BATF were first described to lack T helper cell type 17 (Th17). Th17 cells are a distinct lineage of CD4+ T helper cells that provide protective immunity to infections but are also involved in chronic inflammation and autoimmunity. However, the role of Th17 response in prostate cancer remains controversial. In this study, we took a genetic approach to explore the role of BATF-dependent Th17 cells in prostate cancers by interbreeding BATF-deficient (Batf-/-) mice with mice that are conditionally mutant for PTEN, an established preclinical model for prostate cancer. Batf+/+;Pten-/- and Batf+/-;Pten-/- mice presented similar prostate phenotypes thus were put into one group (named Batf+), which were compared to the Batf-/-;Pten-/- group (named Batf-). We found that Batf- prostates were clearly smaller than Batf+ prostates at 30 weeks. The genitourinary bloc weight was significantly heavier in Batf+ mice than in Batf- mice at 12 and 30 weeks. Batf+ mice developed significantly higher percent of invasive adenocarcinomas in different prostate lobes at 9, 12, and 30 weeks, respectively compared to Batf- mice. Tumors in Batf- mice exhibited lower rates of cellular proliferation and higher apoptosis, as well as reduced inflammatory cell infiltration and angiogenesis in the prostatic stroma. Moreover, Batf- mice exhibited significantly reduced mRNA and protein levels of Th17 cytokines, AP-1 family members and IL-23R, and reduced NF-kB pathway activity in both spleen and prostate tissues. CD4+ T cells from Batf- mice cultured under Th17 conditions for 4 days, expressed reduced AP-1 family-regulated genes compared to Batf+ mice. Prostate cancer cells (LNCaP) with overexpression of BATF in vitro exhibited increased IL-23R expression. Prostate tissues from Batf- mice cultured under IL-6 or Th17 conditions for 4 days had reduced IL-23R expression compared to that from Batf+ mice. Taken together, our results suggest that Batf promotes the initiation and progression of prostate adenocarcinoma via Th17-mediated inflammatory immune responses and that the Batf-IL-23-IL-17 axis is a critical pathway that is needed for prostate adenocarcinoma development and progression. This work was supported by the NIH-National Institute of General Medical Sciences COBRE on Aging and Regenerative Medicine (P20GM103629, PD: S. Michal Jazwinski; PI: Q.Z.) and the Carol Lavin Bernick Faculty Grant from Tulane University (PI: Q.Z.). Citation Format: Qiuyang Zhang, Sen Liu, Bing Zhang. Role of Batf-dependant Th17 immune response in PTEN-null mouse model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1760.

Analysis of Association Between Rs3200401 Long NonCoding RNA MALAT1 Gene Polymorphism and Prostate Adenocarcinoma Development in Ukrainian Population

Today it is well known that protein-coding genes account for only 3% of the total active transcript

Prostate Cancer, Gender Identity, and Testosterone Replacement Therapy in Klinefelter Syndrome: A Case Report and Literature Review

Klinefelter syndrome (KS), karyotype 47, XXY, is a common cause of hypogonadism in males. Patients with this condition often experience symptoms of gonadal failure, which can precipitate gender identity challenges. Treatment with testosterone replacement therapy (TRT) can combat these symptoms by improving sexual function, muscle mass, bone health, and virilization, thereby enhancing the quality of life (QOL). Although TRT is often employed in patients with KS, there is a concern that the application of exogenous testosterone may increase the risk of prostate adenocarcinoma development and progression. We report the case of a 58-year-old male with KS who is also diagnosed with prostate adenocarcinoma and wished to remain on TRT post-radiation therapy in support of his gender identity and QOL. We describe the challenges this patient faced when balancing a rising prostate-specific antigen level and risk of cancer recurrence with his QOL.

Integrative analysis of cancer driver genes in prostate adenocarcinoma.

Large-scale genomics studies have identified recurrently mutated genes in the ETS gene family, including fusions and copy number variations (CNVs), which are involved in the development of prostate adenocarcinoma (PRAD). However, the aetiology of PRAD remains to be fully elucidated. In the present study, 333 driver genes were identified using four computational tools: OncodriveFM, OncodriveCLUST, iCAGES and DrGaP. In addition, 32 driver pathways were identified using DrGaP. SPOP, TP53, SPTA1, AHNAK, HMCN1, ATM, FOXA1, CSMD3, LRP1B and FREM2 were the 10 most recurrently mutated genes in PRAD. ITGAL, TAGAP, SIGLEC10, RAC2 and ITGA4 were the five hub genes in the yellow module that were associated with the number of positive lymph nodes. Hierarchical clustering analysis of the 20 driver genes with the most frequent CNVs revealed three clusters of patients with PRAD. Cluster 3 tumours exhibited significantly higher numbers of positive lymph nodes, higher Gleason scores, more advanced cancer stages and poorer prognosis than cluster 1 and 2 tumours. A total of 48 genes were significantly associated with the number of positive lymph nodes, Gleason scores and pathologic stage in patients with PRAD. The identified set of cancer genes and pathways sheds light on the tumorigenesis of PRAD and creates avenues for the development of prognostic biomarkers and driver gene-targeted therapies in PRAD.

The Role of Chronic Inflammation in Prostate Carcinogenesis: A Follow-Up Study

Purpose To investigate the role of chronic inflammation in prostatic carcinogenesis with an emphasis on cancers of grade group 2 or above. Methods The presence/absence and extent of chronic inflammation and other relevant pathological findings were assessed using prostate needle biopsies obtained from patients with clinical parameters with suspicion of malignancy. In patients with no prior prostate cancer, follow-up biopsies were reviewed and correlated with the initial pathological findings. Results Of 1,006 prostate needle biopsy cases accessioned over 3 years at our institution, the initial biopsies of 244 cases were identified with no evidence of prostate cancer. These cases were divided into two subsets, including 202 cases with and 42 without chronic inflammation. Pathological findings assessed in this subset included post-atrophic hyperplasia, proliferative inflammatory atrophy, and high-grade prostatic intraepithelial neoplasia (HGPIN). Post-atrophic hyperplasia and proliferative inflammatory atrophy were noted only in patients with chronic inflammation. HGPIN was identified in 22 of the initial biopsy cases, specifically in 21 (10.3%) cases with inflammation and one case (2.4%) without inflammation. In follow-up biopsies, 70 patients (34.7%) with chronic inflammation were found to have prostatic adenocarcinoma with Gleason pattern 3 + 3 (42.9%; grade group 1), Gleason pattern 3 + 4(24.3%; grade group 2), Gleason pattern 4 + 3 (10%; grade group 3) and Gleason scores of 8 or higher (22.8%; grade groups 4 or 5), whereas cancer was found in 10 patients (23.8%) without chronic inflammation in the initial biopsy. Patients whose initial biopsies were benign and without inflammation did not show evidence of high-grade cancer (Gleason score of 8 or higher). Closely encroaching inflammation was observed more frequently in cancers of grade group 2 or above (76.5%, 13/17) compared with grade group 1 (50.0%, 3/6). Conclusions Our findings provide additional data supporting a role for chronic inflammation in the development of prostatic adenocarcinoma.

Prognostic value and potential function of splicing events in prostate adenocarcinoma.

Prostate adenocarcinoma (PRAD) is one of the most common types of malignancy in males and at present, effective prognostic indicators are limited. The development of PRAD has been associated with abnormalities in alternative splicing (AS), a requisite biological process of gene expression in eukaryotic cells; however, the prognostic value of AS products and splicing events remains to be elucidated. In the present study, the data of splicing events and the clinical information of PRAD patients were obtained from The Cancer Genome Atlas (TCGA)SpliceSeq and TCGA databases, respectively. A prognostic index (PI) was generated from disease-free survival-associated splicing events (DFS-SEs), which were identified by univariate/multivariate Cox regression analysis. A total of 6,909 DFS-SEs were identified in PRAD. The corresponding genes for the DFS-SEs were significantly enriched in mitochondria and their associated pathways according to Gene Ontology annotation and in the pathways of fatty acid metabolism, oxidative phosphorylation and Huntington's disease according to a Kyoto Encyclopedia of Genes and Genomes pathway analysis. The PI for mutually exclusive exons had the greatest ability to predict the probability of five-year disease-free survival of patients with PRAD, with an area under the time-dependent receiver-operating characteristic curve of 0.7606. Patients with PRAD, when divided into a 'low' and a 'high' group based on their median PI for exon skip values, exhibited a marked difference in disease-free survival (low vs. high, 3,588.45±250.51 vs. 1,531.08±136.50 days; P=7.43×10−9). A correlation network between DFS-SEs of splicing factors and non-splicing factors was constructed to determine the potential mechanisms in PRAD, which included the potential regulatory interaction between the splicing event of splicing factor RNA binding motif protein 5-alternate terminator (AT)-64957 and the splicing event of non-splicing factor heterochromatin protein 1 binding protein 3-AT-939. In conclusion, the PIs derived from DFS-SEs are valuable prognostic factors for patients with PRAD, and the function of splicing events in PRAD deserves further exploration.

Abstract 688: Triphenylstannyl 2-(benzylcarbamoyl)benzoate (Ch-319) modulates AKT/FOXO3a signaling and inhibits progression of prostate cancer

Abstract Prostate cancer (PCa) is the most commonly diagnosed non-cutaneous malignancy in men in the United States. Moreover, its incidence is predicted to surge in many parts of the world due to increase in aging population and change in dietary habits. The PTEN/AKT axis plays a critical role in regulating PCa growth and survival. Synthetic and natural compounds have been explored in preclinical models as potential inhibitors of AKT pathway that can suppress proliferation or induce apoptosis in PCa cells. Triphenyltin compounds are complexes of tin with three phenyl groups attached to hydrocarbon ligands. Structural modifications have been attempted to yield organotins with enhanced cytotoxicity against cancer cells. Here, we synthesized a series of triphenyltin (IV) carboxylate derivatives and evaluated their efficacy against PCa. From this library, triphenylstannyl 2-(benzylcarbamoyl)benzoate (Ch-319) was selected for further studies on the basis of its selective cytotoxicity towards cancer cells. Treatment of PTEN-null PC3M and DU145 PCa cells with Ch-319 (250-500 nM) resulted in reduced viability and induction of cell cycle arrest associated with downregulation of PI3K and dephosphorylation of AKT and PRAS40. In parallel, elevation of FOXO3a expression, activation of pro-apoptotic Bax and downregulation of anti-apoptotic Bcl2 protein was observed. Flowcytometry revealed increased population of apoptotic cells in Ch-319 treated cells. Presensitization of PCa cells with Ch-319 augmented the effect of cabazitaxel, a commonly used taxane in patients with castration-resistant PCa. We next evaluated the effect of Ch-319 in a prostate-specific Pten-/- model where AKT activation is the major driving force for development of prostate adenocarcinoma. Mice treated with Ch-319 (5 µg/animal:intraperitoneally) for 6 weeks showed decreased tumor weights as compared to DMSO treated controls. Histological studies indicated a decrease in the number and size of foci in Ch-319 treated tumors. Consistently, reduced expression of androgen receptor, decreased phosphorylation of AKT and PRAS40 and increase in FOXO3a was observed in Ch-319 treated animals with no signs of organ toxicity. Our studies identify AKT/FOXO3a signaling as a target of triphenyltin (IV) carboxylate derivative Ch-319 and provide a molecular basis of its growth inhibitory effect in PCa cells. We propose that Ch-319 has potential to be developed as an anticancer agent against PCa. Citation Format: Durdana Waseem, Deeba N. Syed, Gul Majid Khan, Ihsan-ul Haq, Hasan Mukhtar. Triphenylstannyl 2-(benzylcarbamoyl)benzoate (Ch-319) modulates AKT/FOXO3a signaling and inhibits progression of 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 688.

Loss of MAOA in epithelia inhibits adenocarcinoma development, cell proliferation and cancer stem cells in prostate.

Monoamine oxidase A (MAOA) is a mitochondrial enzyme, which degrades monoamine neurotransmitters and dietary amines and produces H2O2. Recent studies have shown increased MAOA expression in prostate cancer (PCa), glioma, and classical Hodgkin lymphoma. However, the biological function of MAOA in cancer development remains unknown. In this study, we investigated the role of MAOA in the development of prostate adenocarcinoma by creating a prostate-specific Pten/MAOA knockout (KO) mouse model, in which MAOA-floxP mouse was crossed with the conditional Pten KO PCa mouse that develops invasive PCa. In contrast to Pten KO mice, age-matched Pten/MAOA KO mice exhibited a significant decrease in both prostate size and the incidence of invasive cancer. We observed a significant decline in AKT phosphorylation and Ki67 expression in Pten/MAOA KO mice, which reduced epithelial cell growth and proliferation. As cancer stem cells (CSCs) are required for tumor initiation and growth, we investigated expression of OCT4 and NANOG in the setting of decreased MAOA expression. We found that both OCT4 and NANOG were significantly attenuated in the prostate epithelia of Pten/MAOA KO mice compared to Pten KO mice, which was confirmed with targeted knockdown of MAOA with a short-hairpin(sh) vector targeting MAOA compared to cells transfected with a control vector. Expression of other markers associated with the a stem cell phenotype, including CD44, α2β1, and CD133 as well as HIF-1α+CD44+ stem cells were all decreased in shMAOA PCa cells compared with empty vector-transfected control cells. We also found spheroid formation ability in PCa cells was decreased when endogenous MAOA was suppressed by siRNA or MAOA inhibitor clorgyline in a colony formation assay. Using the TCGA database, elevated MAOA expression was associated with reduced Pten levels in high Gleason grade in patient samples. Further, we found that Pten-positive PCa cells were more resistant to clorgyline treatments than Pten-null cells in tumorigenicity and stemness. Taken together, these studies suggest that MAOA expression promotes PCa development by increasing cell proliferation and CSCs and highlights the potential use of MAOA inhibitors for the treatment of PCa.

COX-2 mediates pro-tumorigenic effects of PKCε in prostate cancer.

The pro-oncogenic kinase PKCε is overexpressed in human prostate cancer and cooperates with loss of the tumor suppressor Pten for the development of prostatic adenocarcinoma. However, the effectors driving PKCε-mediated phenotypes remain poorly defined. Here, using cellular and mouse models, we showed that PKCε overexpression acts synergistically with Pten loss to promote NF-κB activation and induce cyclooxygenase-2 (COX-2) expression, phenotypic traits which are also observed in human prostate tumors. Targeted disruption of PKCε from prostate cancer cells impaired COX-2 induction and PGE2 production. Notably, COX-2 inhibitors selectively killed prostate epithelial cells overexpressing PKCε, and this ability was greatly enhanced by Pten loss. Long-term COX-2 inhibition markedly reduced adenocarcinoma formation as well as angiogenesis in a mouse model of prostate-specific PKCε expression and Pten loss. Overall, our results provide strong evidence for the involvement of the canonical NF-κB pathway and its target gene COX2 as PKCε effectors, and highlight the potential of PKCε as a useful biomarker for the use of COX inhibition for chemopreventive and/or chemotherapeutic purposes in prostate cancer.

δ-Tocopherol inhibits the development of prostate adenocarcinoma in prostate specific Pten-/- mice.

The PTEN/PI3K/AKT axis plays a critical role in regulating cell growth, differentiation and survival. Activation of this signaling pathway is frequently found in human cancers. Our previous studies demonstrated that δ-tocopherol (δ-T) attenuates the activation of AKT by growth factor in prostate cancer cell lines, leading to inhibition of proliferation and induction of apoptosis. Herein, we investigated whether δ-T inhibits the development of prostate adenocarcinoma in prostate-specific Pten-/- (Ptenp-/-) mice in which the activation of AKT is the major driving force for tumorigenesis. By feeding Ptenp-/- mice with AIN93M or 0.2% δ-T supplemented diet starting at the age of 6 or 12 weeks, we found that δ-T treatment reduced prostate adenocarcinoma multiplicity at the age of 40 weeks by 53.3 and 42.7%, respectively. Immunohistochemical (IHC) analysis demonstrated that the phosphorylation of AKT (T308) was reduced in the prostate of the mice administered the δ-T diet. Consistently, proliferation was reduced and apoptosis was increased in prostate lesions of mice on the δ-T diet. Oxidative stress, as determined by IHC staining of 8-OH-dG, was not altered during prostate tumorigenesis, nor was it affected by administration of δ-T. In contrast, α-tocopherol (α-T) at 0.2% in the diet did not affect prostate adenocarcinoma multiplicity in the Ptenp-/- mice. This finding is consistent with data from our previous study that δ-T, but not α-T, inhibits the activation of AKT and the growth of prostate cancer cells. Together, these results demonstrate that δ-T inhibits the development of prostate adenocarcinoma in Ptenp-/- mice, mainly through inhibition of AKT activation.

Abstract 1992: Changes in gene expression of stromal and epithelial cells during prostate cancer progression

Abstract Prostatic intraepithelial neoplasia (PIN) is a histologic abnormality which arises within the secretory epithelium of prostate glands, without disrupting the layer of basal cells which separates the epithelium from the surrounding stroma. PINs are generally considered to be a step towards development of prostatic adenocarcinoma (PCA). Molecular steps in the transition from normal glands to PIN to PCA are poorly understood, in part due to the difficulty of accessing and characterizing individual PINs without contamination from surrounding tissue. Furthermore, stromal cells surrounding pre-cancerous glands may play an active role in cancer development and progression, but are difficult to profile. We utilized the ultra-sensitive and quantitative TempO-Seq gene expression assay of the whole transcriptome to profile microdisections of stromal vs. epithelial cells in archival prostatectomy FFPE tissue. TempO-Seq is highly insensitive to RNA degradation (measurements of intact RNA RIN=9.1 to degraded RNA RIN=3.0 correlate with R2=0.97) and does not require RNA extraction, allowing measurement of both soluble and insoluble cross-linked RNA. This allowed deparaffinization and H&E staining of prostate sections prior to the assay, so that morphology was clearly visible and subsections could be readily extracted and profiled. Microdissections were performed on individual 5 µm FFPE sections of prostatectomies from patients with Gleason sum scores of 6, containing PIN alongside PCA. We removed an ~300 µm diameter region of stroma surrounding individual normal, PIN, and PCA glands, followed by targeted dissection of the gland epithelium itself which was lysed and assayed separately. Stroma and epithelial samples from 3 adjacent sections of the same glands were pooled together (adding up to ~0.005 mm3 tissue/replicate), and then profiled. We compared the whole transcriptome expression profile of stroma vs. epithelium at every stage of progression. Our results indicate that stroma and epithelium undergo separate and distinct changes in gene expression patterns throughout the development of PCA, most notably in genes involved in cell-cell signaling. Specifically, we found that changes in cAMP signaling in stromal cells accompanied the shift from normal glands to PIN. Furthermore, changes in androgen signaling pathways were present in stromal cells as well as in epithelium during the transition from PIN to invasive carcinoma. These results shed light on the molecular progression to PCA and demonstrate the feasibility of profiling gene expression at the level of tissue microstructure to elucidate changes within the tissue microenvironment. Since archival FFPE can be used, studies of progression can be carried out and the findings translated to diagnosis and prognosis as well as to the investigation of new therapeutic approaches. Citation Format: Milos Babic, Elliot Imler, Peter Shepard, Joanne Yeakley, Raymond Nagle, Bruce Seligmann. Changes in gene expression of stromal and epithelial cells during prostate cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1992. doi:10.1158/1538-7445.AM2017-1992

Abstract 5247: δ-Tocopherol inhibits the development of prostate adenocarcinoma in prostate specific Pten-/- mice

Abstract PTEN/PI3K/AKT axis plays a critical role in regulating cell growth, metabolism, migration, differentiation, and survival. Activation of this signal pathway is frequently found in human cancers. Data from our previous studies demonstrated that δ-tocopherol (δ-T) attenuates the activation of AKT by growth factor in prostate cancer cell lines, leading to inhibiting proliferation and inducing apoptosis. Herein, we investigated whether δ-T is able to inhibit the development of prostate adenocarcinoma in prostate-specific Pten-/- (Ptenp-/-) mice, in which the activation of AKT resulted from loss of Pten is the driver. By feeding Ptenp-/- mice with the AIN93M or 0.2% δ-T supplemented diet starting at the age of 6 or 12 weeks, we found that, at the age of 40 weeks, δ-T treatment reduced the number of invasive prostate adenocarcinoma by 53.3% (p<0.005) and 42.7% (p<0.001), respectively. By immunohistochemical analysis, the activation of AKT via phosphorylation of AKT(T308) was found reduced in the prostate of the mice on δ-T diet. Consistently, proliferation was reduced and apoptosis was increased in prostate of the mice on δ-T diet. We also determined oxidative stress by immunohistochemical staining of 8-OH-dG and nitro-tyrosine, and found that these markers were not altered during prostate tumorigenesis, nor they were affected by δ-T. A 0.2% α-tocopherol diet was also used to feed Ptenp-/- mice, but did not inhibit the development of prostate adenocarcinoma, consistent with previous data demonstrating that α-tocopherol did not inhibit the activation of AKT. Together, these results support that δ-T is effective in supressing the development of prostate adenocarcinoma in Ptenp-/- mice through inhibition of AKT activation. Citation Format: Xu Yang, Wang Hong, Guocan Wang, Anna Liu, Maarten c. Bosland, Chung s. Yang. δ-Tocopherol inhibits the development of prostate adenocarcinoma in prostate specific Pten-/- mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5247. doi:10.1158/1538-7445.AM2017-5247

Yes-Associated Protein Expression Is Correlated to the Differentiation of Prostate Adenocarcinoma.

BackgroundYes-associated protein (YAP) in the Hippo signaling pathway is a growth control pathway that regulates cell proliferation and stem cell functions. Abnormal regulation of YAP was reported in human cancers including liver, lung, breast, skin, colon, and ovarian cancer. However, the function of YAP is not known in prostate adenocarcinoma. The purpose of this study was to investigate the role of YAP in tumorigenesis, differentiation, and prognosis of prostate adenocarcinoma.MethodsThe nuclear and cytoplasmic expression of YAP was examined in 188 cases of prostate adenocarcinoma using immunohistochemistry. YAP expression levels were evaluated in the nucleus and cytoplasm of the prostate adenocarcinoma and the adjacent normal prostate tissue. The presence of immunopositive tumor cells was evaluated and interpreted in comparison with the patients’ clinicopathologic data.ResultsYAP expression levels were not significantly different between normal epithelial cells and prostate adenocarcinoma. However, YAP expression level was significantly higher in carcinomas with a high Gleason grades (8–10) than in carcinomas with a low Gleason grades (6–7) (p < .01). There was no statistical correlation between YAP expression and stage, age, prostate-specific antigen level, and tumor volume. Biochemical recurrence (BCR)–free survival was significantly lower in patients with high YAP expressing cancers (p = .02). However high YAP expression was not an independent prognostic factor for BCR in the Cox proportional hazards model.ConclusionsThe results suggested that YAP is not associated with prostate adenocarcinoma development, but it may be associated with the differentiation of the adenocarcinoma. YAP was not associated with BCR.

MP33-14 THE ROLE OF CHRONIC PROSTATIC INFLAMMATION IN PROSTATE CARCINOGENESIS: A RETROSPECTIVE FOLLOW-UP STUDY

You have accessJournal of UrologyProstate Cancer: Detection & Screening III1 Apr 2017MP33-14 THE ROLE OF CHRONIC PROSTATIC INFLAMMATION IN PROSTATE CARCINOGENESIS: A RETROSPECTIVE FOLLOW-UP STUDY Liwei Jia, Shawn Silver, and Gregory MacLennan Liwei JiaLiwei Jia More articles by this author , Shawn SilverShawn Silver More articles by this author , and Gregory MacLennanGregory MacLennan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.1010AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Chronic inflammation is a well-known contributor to several human cancers. The aim of this study was to investigate the role of chronic prostatic inflammation in prostatic carcinogenesis, with a specific focus on high grade cancers. METHODS A retrospective search was performed to identify prostate biopsy cases accessioned in our institution between 01/01/2007 and 12/31/2009. Pathology reports of each case were reviewed. We reviewed the slides of all patients with benign initial prostate biopsies, followed by at least one set of prostate biopsies longer than one year after the initial biopsies. The presence and extent of chronic inflammation and other pathological findings were assessed and recorded. We defined chronic inflammation as involvement of 1% or more of the surface area of the prostatic tissue by a chronic inflammatory cell infiltrate. We also recorded the presence or absence of postatrophic hyperplasia (PAH) and inflamed atrophy (IA), a lesion defined by the presence of clusters of atrophic glands surrounded by a prominent and readily identifiable lymphocytic infiltrate. Cases in which prostatic adenocarcinoma was subsequently diagnosed were evaluated for Gleason pattern and score. RESULTS Among 1006 patients who underwent prostate biopsies, a total of 244 cases with available slides were diagnosed initially as benign prostatic tissue, of which two patient subsets were identified, including 206 patients with and 38 patients without chronic inflammation in the initial biopsy. Post-atrophic hyperplasia (PAH) and inflamed atrophy (IA) were more frequently identified in patients with chronic inflammation. In follow up biopsies performed at least 1 year following the initial benign biopsies, 75 patients (36%) with chronic inflammation were found to have adenocarcinoma, whereas cancer was found in only 6 patients (16%) without chronic inflammation in the initial biopsies. Of those who were found to have cancer after the initial biopsies showing only chronic inflammation, 46% had a Gleason 3 + 3 pattern (grade group 1), 20% had a Gleason 3 + 4 pattern (grade group 2), and 13% had a Gleason 4 + 3 pattern (grade group 3). Significantly, high grade cancers with Gleason score of 8 or higher (grade groups 4 or 5) were identified in 21% (16 of 75) of patients with chronic inflammation in the initial benign biopsies, whereas no patient without chronic inflammation in the initial benign biopsies were found to have cancer with a Gleason score of 8 or higher (grade groups 4 or 5). CONCLUSIONS Our retrospective follow up study suggests a strong association between chronic prostatic inflammation and the development of prostatic adenocarcinoma. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e422 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Liwei Jia More articles by this author Shawn Silver More articles by this author Gregory MacLennan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Copper as a target for prostate cancer therapeutics: copper-ionophore pharmacology and altering systemic copper distribution.

Copper-ionophores that elevate intracellular bioavailable copper display significant therapeutic utility against prostate cancer cells in vitro and in TRAMP (Transgenic Adenocarcinoma of Mouse Prostate) mice. However, the pharmacological basis for their anticancer activity remains unclear, despite impending clinical trails. Herein we show that intracellular copper levels in prostate cancer, evaluated in vitro and across disease progression in TRAMP mice, were not correlative with copper-ionophore activity and mirrored the normal levels observed in patient prostatectomy tissues (Gleason Score 7 & 9). TRAMP adenocarcinoma cells harbored markedly elevated oxidative stress and diminished glutathione (GSH)-mediated antioxidant capacity, which together conferred selective sensitivity to prooxidant ionophoric copper. Copper-ionophore treatments [CuII(gtsm), disulfiram & clioquinol] generated toxic levels of reactive oxygen species (ROS) in TRAMP adenocarcinoma cells, but not in normal mouse prostate epithelial cells (PrECs). Our results provide a basis for the pharmacological activity of copper-ionophores and suggest they are amendable for treatment of patients with prostate cancer. Additionally, recent in vitro and mouse xenograft studies have suggested an increased copper requirement by prostate cancer cells. We demonstrated that prostate adenocarcinoma development in TRAMP mice requires a functional supply of copper and is significantly impeded by altered systemic copper distribution. The presence of a mutant copper-transporting Atp7b protein (tx mutation: A4066G/Met1356Val) in TRAMP mice changed copper-integration into serum and caused a remarkable reduction in prostate cancer burden (64% reduction) and disease severity (grade), abrogating adenocarcinoma development. Implications for current clinical trials are discussed.