Human Benign Prostatic Hyperplasia Tissue Research Articles

A Comparative Study of Histotripsy Parameters for the Treatment of Fibrotic ex-vivo Human Benign Prostatic Hyperplasia Tissue.

Histotripsy is a noninvasive focused ultrasound therapy that mechanically fractionates tissue to create well-defined lesions. In a previous clinical pilot trial to treat benign prostatic hyperplasia (BPH), histotripsy did not result in consistent objective improvements in symptoms, potentially because of the fibrotic and mechanically tough nature of this tissue. In this study, we aimed to identify the dosage required to homogenize BPH tissue by different histotripsy modalities, including boiling histotripsy (BH) and cavitation histotripsy (CH). A method for histotripsy lesion quantification via entropy (HLQE) analysis was developed and utilized to quantify lesion area of the respective treatments. These data were correlated to changes in mechanical stiffness measured by ultrasound shear-wave elastography before and after treatment with each parameter set and dose. Time points corresponding to histologically observed complete lesions were qualitatively evaluated and quantitatively measured. For the BH treatment, complete lesions occurred with >=30s treatment time, with a corresponding maximum reduction in stiffness of -90.9±7.2(s.d.)%. High pulse repetition frequency (PRF) CH achieved a similar reduction to that of BH at 288s (-91.6±6.0(s.d.)%), and low-PRF CH achieved a (-82.1±5.1(s.d.)%) reduction in stiffness at dose >=144s. Receiver operating characteristic curve analysis showed that a >~75% reduction in stiffness positively correlated with complete lesions observed histologically, and can provide an alternative metric to track treatment progression.

The activated complement pathway in the fibrous process of benign prostatic hyperplasia.

To elucidate the changes in activated complement pathway in the fibrous process of benign prostatic hyperplasia (BPH), we analyzed the correlation between complement component expression and histological types of fibrosis using human BPH tissue. Fifty-six histological BPH patients who underwent prostate needle biopsy at our institution (mean age 68.6 ± 6.5 years), divided into two histological groups, fibromuscular and fibrous, were compared. Inflammatory cell infiltration in BPH tissue was evaluated by immunohistochemical staining using CD45, with complement expression analysis performed using C3, factor B, and C5b-9 antibody, and the occupancy ratio of the stained region was calculated. Further, correlation between the histological types of fibrous components in BPH tissue and lower urinary tract symptoms questionnaires was analyzed. Twenty-seven (48.2%) and 29 (51.8%) cases were classified in the fibromuscular and fibrous groups, respectively. The proportion of CD45-positive cells in BPH tissue was significantly higher in the fibromuscular group. In complement component analysis, factor B did not significantly differ between groups, while C3 (fibromuscular group; 10.7 ± 8.2%, fibrous group; 16.4 ± 12.7%) and C5b-9 (fibromuscular group; 15.9 ± 6.2%, fibrous group; 17.6 ± 9.2%) were significantly higher in the fibrous group (p = 0.04, p = 0.04, respectively). International Prostate Symptom Score Q5 subscore, indicating slow stream, was significantly higher in the fibrous group (p = 0.04). In fibrous BPH with abundant fibrosis, the late complement pathway in addition to alternative pathway was activated compared to fibromuscular BPH. These results suggested that the alternative and late complement pathways were involved in the histological fibrous process of BPH.

Therapeutic potential of pomegranate juice-derived nanovesicles in nude mouse benign prostatic hyperplasia (BPH) xenograft model

Benign prostatic hyperplasia (BPH) and associated lower urinary tract symptoms affect a large percentage of the male population and places a substantial burden on the world health system. Current therapies include 5-alpha reductase inhibitors and alpha-blockers that are only partially effective and pose a huge economic burden, emphasizing the urgent need for effective, economical therapies. We isolated nanovesicles from pomegranate juice (Punica Granatum) (referred to as ‘POM-NVs’) and report to our knowledge for the first time, that these vesicles possess therapeutic potential against BPH. Following extensive characterization of POM-NVs, we tested their therapeutic potential in vitro using BPH1 cell line and identified a potential anti-proliferative and pro-apoptotic effect. We further tested these vesicles using a clinically relevant xenograft mouse BPH model derived from human BPH tissues. Remarkably, POM-NVs could reverse the BPH phenotype conferred by TGF-β mediated signaling and induced epithelial-to-mesenchymal (EMT) reversal, leading to the restoration of prostate epithelial states in vivo and in vitro. Furthermore, these vesicles attenuated bone morphogenic protein 5 (BMP5) signaling, a cardinal alteration that is instrumental in driving BPH. Considering the large incidences of BPH and its associated economic burdens, our study has important implications and can potentially improve the clinical management of BPH.

Roles of NF-κB activation in benign prostatic hyperplasia and association between NF-κB and HIF-1α

ObjectiveThe aim of this study was to elucidate the role of NF-κB activation in benign prostatic hyperplasia (BPH) using immunohistochemistry. MethodsImmunohistochemical staining for NF-κB was performed and evaluated, dividing into glands and stroma in 101 human BPH tissues. To evaluate the impacts of NF-κB activation on BPH progression, correlations between NF-κB expression and clinical findings, hormone receptors, and HIF-1α were evaluated. ResultsNF-κB expression was found in 37.6% and 30.7% in glands and stroma of BPH, respectively. Total and T-zone volumes in transrectal ultrasonography were significantly higher in patients with NF-κB activation than those without NF-κB activation in the stroma. However, NF-κB activation of stroma was not correlated with HIF-1α expression and microvessel density. In subgroup analysis based on NF-κB activation, androgen and progesterone receptors of stroma were highly expressed in HIF-1α negative cases than in HIF-1α positive cases. In cases without NF-κB activation, patients with HIF-1α positivity showed a high frequency of diffuse fibrosis than those with HIF-1α negativity (P = 0.001). ConclusionTaken together, our result showed that NF-κB activation of stroma was significantly correlated with low total and T-zone volumes in transrectal ultrasonography. Diffuse fibrosis was frequently found in patients with NF-κB inactivation and HIF-1α positivity.

PD16-06 SHIFT CHANGE OF THE ACTIVATED COMPLEMENT PATHWAY IN THE FIBROTIC PROCESS ASSOCIATED WITH PROGRESSION OF BENIGN PROSTATIC HYPERPLASIA

You have accessJournal of UrologyCME1 May 2022PD16-06 SHIFT CHANGE OF THE ACTIVATED COMPLEMENT PATHWAY IN THE FIBROTIC PROCESS ASSOCIATED WITH PROGRESSION OF BENIGN PROSTATIC HYPERPLASIA Junya Hata, Yusuke Kirihana, Tomoyuki Kumekawa, Hiroki Natsuya, Yusuke Hakozaki, Syunsuke Yoshioka, Hitomi Imai, Kanako Matsuoka, Tomoyuki Koguchi, Yuichi Sato, Hidenori Akaihata, Masao Kataoka, and Yoshiyuki Kojima Junya HataJunya Hata More articles by this author , Yusuke KirihanaYusuke Kirihana More articles by this author , Tomoyuki KumekawaTomoyuki Kumekawa More articles by this author , Hiroki NatsuyaHiroki Natsuya More articles by this author , Yusuke HakozakiYusuke Hakozaki More articles by this author , Syunsuke YoshiokaSyunsuke Yoshioka More articles by this author , Hitomi ImaiHitomi Imai More articles by this author , Kanako MatsuokaKanako Matsuoka More articles by this author , Tomoyuki KoguchiTomoyuki Koguchi More articles by this author , Yuichi SatoYuichi Sato More articles by this author , Hidenori AkaihataHidenori Akaihata More articles by this author , Masao KataokaMasao Kataoka More articles by this author , and Yoshiyuki KojimaYoshiyuki Kojima More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002548.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: In a previous study, we reported that complement activation by an autoimmune reaction was associated with the growth process of benign prostatic hyperplasia (BPH). On the other hand, BPH was reported to show a histological transition from smooth muscle-dominant type to fibrous-dominant type as it becomes more severe. Therefore, we hypothesized that activation of complement pathways might be associated with the fibrous process of BPH with progression. In this study, to clarify the activated complement pathway in the fibrous process of BPH with progression, we analyzed the histological severity and the expression of complement components using human fibrous BPH tissues. METHODS: The subjects were 56 histological BPH patients who underwent prostate needle biopsy due to high PSA levels in our institutions (mean age 68.6 ± 6.5 years). BPH patients were divided into two histological groups, fibromuscular type and fibrous type, by hematoxylin-eosin and elastica-Masson staining. Complement expression function analysis was performed by immunohistochemical staining using C3, factor B, and C5b-9 antibody, and the occupancy ratio of the stained region was calculated. RESULTS: Twenty-seven cases (48.2%) were classified as fibromuscular type, and 29 cases (51.8%) were classified as fibrous type. The proportion of fibrous components was significantly lower in the fibromuscular type than in the fibrous type (fibromuscular type 36.0 ± 12.9%, fibrous type 61.1 ± 11.7% (p<0.01)). In the expression analyses of complement components, factor B was not significantly different between the two groups, whereas C3 (fibromuscular type 10.7 ± 8.2%, fibrous type 16.4 ± 12.7%) and C5b-9 (fibromuscular type 15.9 ± 6.2%, fibrous type 17.6 ± 9.2%) were significantly upregulated in fibrous type compared to fibromuscular type (p = 0.04, p = 0.04). Furthermore, abundant infiltration of inflammatory cells was observed in the fibromuscular type (p = 0.03). CONCLUSIONS: In the fibrous process of BPH from fibromuscular type to fibrous type histologically, the inflammatory amplification loop enhancement by complement alternative pathway activation promotes the fibrosis of BPH, and finally shifts to fibrotic BPH. These results suggested that shift change to complement late pathway activation was involved in the progression of fibrous BPH. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e271 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Junya Hata More articles by this author Yusuke Kirihana More articles by this author Tomoyuki Kumekawa More articles by this author Hiroki Natsuya More articles by this author Yusuke Hakozaki More articles by this author Syunsuke Yoshioka More articles by this author Hitomi Imai More articles by this author Kanako Matsuoka More articles by this author Tomoyuki Koguchi More articles by this author Yuichi Sato More articles by this author Hidenori Akaihata More articles by this author Masao Kataoka More articles by this author Yoshiyuki Kojima More articles by this author Expand All Advertisement PDF DownloadLoading ...

MP04-02 ANALYSIS OF EX-VIVO HUMAN BPH TISSUE TREATED BY HISTOTRIPSY: DOSE AND PARAMETER EFFECTS

You have accessJournal of UrologyCME1 May 2022MP04-02 ANALYSIS OF EX-VIVO HUMAN BPH TISSUE TREATED BY HISTOTRIPSY: DOSE AND PARAMETER EFFECTS Zorawar Singh, Yashwanth N. Kumar, Yak-Nam Wang, Adam Maxwell, Matthew Bruce, Rishi Sekar, Eli Vlaisavljevich, Lawrence D. True, and George R. Schade Zorawar SinghZorawar Singh More articles by this author , Yashwanth N. KumarYashwanth N. Kumar More articles by this author , Yak-Nam WangYak-Nam Wang More articles by this author , Adam MaxwellAdam Maxwell More articles by this author , Matthew BruceMatthew Bruce More articles by this author , Rishi SekarRishi Sekar More articles by this author , Eli VlaisavljevichEli Vlaisavljevich More articles by this author , Lawrence D. TrueLawrence D. True More articles by this author , and George R. SchadeGeorge R. Schade More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002521.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Histotripsy is a promising alternative to more invasive procedures for the treatment of benign prostatic hyperplasia (BPH). However, in a phase 1 clinical trial, histotripsy failed to provide objective improvements in urinary function. One explanation may be that the clinical trial pulse parameters, optimized in a canine model, do not translate to more fibrotic human prostates. We aimed to evaluate the efficacy of different histotripsy parameter settings to identify a more efficacious exposure in ex-vivo human BPH tissue (EVHP). METHODS: Deidentified EVHP tissue specimens (n=17) were treated with an 18 element 700kHz cavitation histotripsy transducer, which approximated the device used in the prior clinical trial, and a 1.5MHz boiling histotripsy (BH) transducer. Three different histotripsy exposures were compared: 1) a high pulse repetition frequency (PRF) (approximating the clinical system cavitation cloud histotripsy [CCH]), 2) low PRF CCH and 3) 1Hz PRF BH. Treatment effectiveness was evaluated by measuring pre vs. post treatment Young’s modulus through shear wave elastography (SWE) and histologic analysis (H&E, Masson’s Trichome) of the targeted area. The dose was standardized at 30s/mm and escalated as required to note a significant change in SWE. RESULTS: At the given dose of 30s/mm, BH treatments resulted in a significant 40.1% reduction in SWE Young’s modulus (Figure 1A) p < 0.001. At this dose, neither of the CCH parameter settings (500Hz and 10Hz) resulted in a significant reduction in the Young’s modulus with % changes of differences of 3.1% (p = .83) and -5% (p = .08), respectively. For CCH settings, greater exposure times (up to 90s) did result in significant stiffness reduction of 12.5% (p = 0.04) and 41.1% (p < 0.001) for 500Hz and 10Hz parameter settings, respectively. On histology (Figure 1B), at 30s/mm, the CCH parameter settings resulted in minimal intermittent damage while BH treatments caused near complete ablation. At increasing doses, CCH was also capable of causing complete liquefactive cavity formation. CONCLUSIONS: BH parameters applied in this study resulted in more complete ablation of EVH tissue in comparison to CCH parameters. Additionally, histotripsy treatment parameters similar to those applied with a previous clinical system did not produce consistent ablation of EVHP possibly explaining trial results. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e56 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Zorawar Singh More articles by this author Yashwanth N. Kumar More articles by this author Yak-Nam Wang More articles by this author Adam Maxwell More articles by this author Matthew Bruce More articles by this author Rishi Sekar More articles by this author Eli Vlaisavljevich More articles by this author Lawrence D. True More articles by this author George R. Schade More articles by this author Expand All Advertisement PDF DownloadLoading ...

The Roles of ZnT1 and ZnT4 in Glucose-Stimulated Zinc Secretion in Prostate Epithelial Cells.

We have previously demonstrated by MRI that high glucose stimulates efflux of zinc ions from the prostate. To our knowledge, this phenomena had not been reported previously and the mechanism remains unknown. Here, we report some initial observations that provide new insights into zinc processing during glucose-stimulated zinc secretion (GSZS) in the immortalized human prostate epithelial cell line, PNT1A. Additionally, we identified the subtypes of zinc-containing cells in human benign prostatic hyperplasia (BPH) tissue to further identify which cell types are likely responsible for zinc release in vivo. An intracellular fluorescence marker, FluoZin-1-AM, was used to assess the different roles of ZnT1 and ZnT4 in zinc homeostasis in wild type (WT) and mRNA knockdown PNT1A cell lines. Additionally, Bafilomycin A1 (Baf) was used to disrupt lysosomes and assess the role of lysosomal storage during GSZS. ZIMIR, an extracellular zinc-responsive fluorescent marker, was used to assess dynamic zinc efflux of WT and ZnT1 mRNA knockdown cells exposed to high glucose. Electron microscopy was used to assess intracellular zinc storage in response to high glucose and evaluate how Bafilomycin A1 affects zinc trafficking. BPH cells were harvested from transurtheral prostatectomy tissue and stained with fluorescent zinc granule indicator (ZIGIR), an intracellular zinc-responsive fluorescent marker, before being sorted for cell types using flow cytometry. Fluorescent studies demonstrate that ZnT1 is the major zinc efflux transporter in prostate epithelial cells and that loss of ZnT1 via mRNA knockdown combined with lysosomal storage disruption results in a nearly 4-fold increase in cytosolic zinc. Knockdown of ZnT1 dramatically reduces zinc efflux during GSZS. Electron microscopy (EM) reveals that glucose stimulation significantly increases lysosomal storage of zinc; disruption of lysosomes via Baf or ZnT4 mRNA knockdown increases multi-vesicular body (MVB) formation and cytosolic zinc levels. In human BPH tissue, only the luminal epithelial cells contained significant amounts of zinc storage granules. Exposure of prostate epithelial cells to high glucose alters zinc homeostasis by inducing efflux of zinc ions via ZnT1 channels and increasing lysosomal storage via ZnT4. Given that prostate cancer cells undergo profound metabolic changes that result in reduced levels of total zinc, understanding the complex interplay between glucose exposure and zinc homeostasis in the prostate may provide new insights into the development of prostate carcinogenesis.

Morphological change and characteristics of myofibroblasts during the growth process of benign prostatic hyperplasia.

To clarify the morphological change and characteristics of myofibroblast during the growth process of benign prostatic hyperplasia. This study examined the characteristics of myofibroblasts during the growth process of the prostate in the stromal component-dominant benign prostatic hyperplasia rat model. Transforming growth factor-β1 and insulin-like growth factor-binding protein3 expression were evaluated by western blotting (n=6). We used double immunohistochemical staining to evaluate the number of myofibroblasts positive for α-smooth muscle actin and vimentin in benign prostatic hyperplasia tissues. Expression and histological analyses of the benign prostatic hyperplasia were also carried out in rats at 2, 3 and 8weeks after urogenital sinus implantation (n=6). To evaluate the fine morphological characteristics of myofibroblasts in human benign prostatic hyperplasia tissues, electron microscopy analysis was additionally carried out. There was a significant upregulation of the transforming growth factor-β1 and insulin-like growth factor-binding protein3 expression in benign prostatic hyperplasia (P<0.05). There was a significant increase in the number of myofibroblasts in benign prostatic hyperplasia (P<0.05) compared with normal prostate, with these abundantly located in the stromal area. The transforming growth factor-β1 and insulin-like growth factor-binding protein3 expression and number of myofibroblasts showed a time-dependent increase (P<0.05), with growth factor expressions preceding the myofibroblast increase. Electron microscopy confirmed that the myofibroblast progenitor cells, which possess abundant stress fibers, were predominantly located around fibrous areas in human benign prostatic hyperplasia. Differentiation into myofibroblasts induced by transforming growth factor-β1 and insulin-like growth factor-binding protein3 actively occurs during the growth process of benign prostatic hyperplasia. Myofibroblast progenitor cells seem to be associated with prostatic fibrosis in human benign prostatic hyperplasia.

Complement activation by autoantigen recognition in the growth process of benign prostatic hyperplasia

The pathophysiology of benign prostatic hyperplasia (BPH) remained unclear. Here, we concentrated on the complement activation in the growth of BPH using a rat model. BPH tissues were harvested from rats after rat urogenital sinus implantation. The local expression and deposition levels of C1q, C3, mannose-binding lectin (MBL), factor B (FB), and C5b-9 in the rat and human BPH tissues were analyzed by real-time RT-PCR, western blotting and immunohistochemistry (IHC). Serum IgG levels in the rat BPH model were analyzed by ELISA, and IHC was used to assess tissue localization. Proteins binding serum IgG autoantibody in the BPH rats were isolated by immunoprecipitation. C1q, C3, MBL, FB and C5b-9 were highly localized in rat BPH tissues compared to normal tissues. In contrast, C3, FB and C5b-9, but not C1q and MBL, were abundantly detected in human BPH tissues compared to normal tissues. Diffuse localization of IgG in rat BPH tissues was found. Heat shock protein 90, annexin, α-smooth muscle actin, and β-actin were identified as targets for IgG autoantibodies in the BPH model. Our results strongly suggested the role for complement activation in the growth process of BPH, likely triggered by classical pathway activation with autoantibodies.

Infiltrating mast cells enhance benign prostatic hyperplasia through IL-6/STAT3/Cyclin D1 signals.

Early evidences have showed that mast cells could infiltrate into benign prostatic hyperplasia (BPH) tissues, but the exact role of mast cells in BPH development remains unclear. In this study, we identified more mast cells existing in human BPH tissues compared with that in the normal prostate. In the in vitro co-culture system, BPH-1 prostate cells promoted activation and migration of mast cells, and mast cells conversely stimulated BPH-1 cells proliferation significantly. Molecular analysis demonstrated that mast cell-derived interleukin 6 (IL-6) could activate STAT3/Cyclin D1 signals in BPH-1 cells. Blocking IL-6 or STAT3 partially reverse the capacity of mast cells to enhance BPH-1 cell proliferation. Our findings suggest that infiltrating mast cells in BPH tissues could promote BPH development via IL-6/STAT3/Cyclin D1 signals. Therefore, targeting infiltrating mast cells may improve the therapeutic effect of BPH.

MP17-04 COMPLEMENT ACTIVATION MECHANISM ACTIVATED BY AUTOANTIGEN RECOGNITION DURING GROWTH OF BENIGN PROSTATIC HYPERPLASIA

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2017MP17-04 COMPLEMENT ACTIVATION MECHANISM ACTIVATED BY AUTOANTIGEN RECOGNITION DURING GROWTH OF BENIGN PROSTATIC HYPERPLASIA Junya Hata, Kanako Matsuoka, Yuichi Sato, Hidenori Akaihata, Masao Kataoka, Soichiro Ogawa, Nobuhiro Haga, Kei Ishibashi, Ken Aikawa, and Yoshiyuki Kojima Junya HataJunya Hata More articles by this author , Kanako MatsuokaKanako Matsuoka More articles by this author , Yuichi SatoYuichi Sato More articles by this author , Hidenori AkaihataHidenori Akaihata More articles by this author , Masao KataokaMasao Kataoka More articles by this author , Soichiro OgawaSoichiro Ogawa More articles by this author , Nobuhiro HagaNobuhiro Haga More articles by this author , Kei IshibashiKei Ishibashi More articles by this author , Ken AikawaKen Aikawa More articles by this author , and Yoshiyuki KojimaYoshiyuki Kojima More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.593AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The association between the pathogenesis of benign prostatic hyperplasia (BPH) and inflammation has recently received attention. We previously showed that not only the inflammation response pathway, but also the classical complement pathway is activated in BPH tissue from model rats with stroma-dominant BPH. The classical complement pathway is activated by autoantigens that recognize immune complexes and it is responsible for various diseases via a mechanism that amplifies inflammation. We postulated that immune complexes amplify inflammation through complement activation, which leads to prostatic proliferation. Therefore, we expressed complement factors, analyzed their functions, and identified autoantigens to understand the pathogenic mechanism of BPH. METHODS Fetal urogenital sinus (UGS) isolated from male 20-day-old rat embryos was implanted into the ventral prostate of pubertal male rats to create rat models of BPH. Complement factors were expressed and functionally analyzed in BPH tissues, and then serum concentrations of IgG and the expression of complement factors in BPH tissues were assessed. We immunoprecipitated BPH protein using an anti-IgG antibody to identify antigens, and analyzed the protein by mass spectrometry after SDS-PAGE separation. The expression of complement factors in human BPH tissue was also analyzed. RESULTS The expression of complement factors C1q, C3, MBL, factor B, and MAC was significantly up-regulated in tissues from BPH rats compared with those from normal rats (p<0.01). The classical complement pathway was initially activated, followed by an alternative complement pathway activated in BPH. These complement factors were also up-regulated mostly in stromal areas of human BPH. The serum IgG concentration was significantly increased (398.1 ng/mL, p<0.01) in rat BPH and IgG was deposited in stromal areas of the BPH. Mass spectrometry of IgG binding protein identified annexin, Hsp90, and β-actin as antigens of immunocomplexes. CONCLUSIONS We clarified that the immune system is responsible for the development of BPH. Complement pathway activation by immunocomplexes recognizing annexin, Hsp90, and β-actin as autoantigens might be responsible for the pathogenesis of BPH. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e213 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Junya Hata More articles by this author Kanako Matsuoka More articles by this author Yuichi Sato More articles by this author Hidenori Akaihata More articles by this author Masao Kataoka More articles by this author Soichiro Ogawa More articles by this author Nobuhiro Haga More articles by this author Kei Ishibashi More articles by this author Ken Aikawa More articles by this author Yoshiyuki Kojima More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Aberrant Transforming Growth Factor-β Activation Recruits Mesenchymal Stem Cells During Prostatic Hyperplasia.

Benign prostatic hyperplasia (BPH) is the overgrowth of prostate tissues with high prevalence in older men. BPH pathogenesis is not completely understood, but it is believed to be a result of de novo overgrowth of prostatic stroma. In this study, we show that aberrant activation of transforming growth factor‐β (TGF‐β) mobilizes mesenchymal/stromal stem cells (MSCs) in circulating blood, which are recruited for the prostatic stromal hyperplasia. Elevated levels of active TGF‐β were observed in both a phenylephrine‐induced prostatic hyperplasia mouse model and human BPH tissues. Nestin lineage tracing revealed that 39.6% ± 6.3% of fibroblasts and 73.3% ± 4.2% smooth muscle cells were derived from nestin+ cells in Nestin‐Cre, Rosa26‐YFPflox/+mice. Nestin+ MSCs were increased in the prostatic hyperplasia mice. Our parabiosis experiment demonstrate that nestin+ MSCs were mobilized and recruited to the prostatic stroma of wild‐type mice and gave rise to the fibroblasts. Moreover, injection of a TGF‐β neutralizing antibody (1D11) inhibits mobilization of MSCs, their recruitment to the prostatic stroma and hyperplasia. Importantly, knockout of TβRII in nestin+ cell lineage ameliorated stromal hyperplasia. Thus, elevated levels of TGF‐β‐induced mobilization and recruitment of MSCs to the reactive stroma resulting in overgrowth of prostate tissues in BPH and, thus, inhibition of TGF‐β activity could be a potential therapy for BPH. Stem Cells Translational Medicine 2017;6:394–404

The Differential Expression and Function of the Inflammatory Chemokine Receptor CXCR5 in Benign Prostatic Hyperplasia and Prostate Cancer.

Background: Chemokine and chemokine receptors could have played an important role in tumor angiogenesis and distant metastasis. The mechanism of inflammation, expression and function of chemokines and chemokine receptors in benign prostatic hyperplasia (BPH) and prostate cancer (PCa) remain unclear. The purpose of present study is to detect differential expression and function of chemokines and chemokine receptors (CCRs) in BPH and PCa.Methods: BPH-1 and peripheral blood mononuclear cells (PBMCs) were co-cultured in Transwell chambers, and human normal prostate (NP) tissues, BPH tissues and PCa tissues were collected. CCR gene-chips were used to analyze and compare the differential expression of CCRs in BPH-1 cells, BPH-1 cells co-cultured with PBMCs, and LNCaP cells. The differential expression of CCRs was detected and validated using real-time PCR, western blotting and immunofluorescence (IF). The proliferation of LNCaP cells was also investigated after the knockdown CXCR5.Results: Results of gene-chips indicated that there was low or no expression of CCR10, CXCR1, CXCR3 and CXCR5 in BPH-1 cells, whereas the expression of these receptors in BPH-1 cells was increased by PBMCs, and the expression was high in LNCaP cells. Furthermore, real-time PCR and western blotting confirmed the above mentioned results. IF verified no or low expression of CXCR1, CXCR3 and CXCR5 in NP tissues, low or moderate expression in BPH and high expression in PCa. However, CCR10 was not expressed at detectable levels in the three groups. The growth and proliferation of LNCaP cells was markedly inhibited after down-regulation of CXCR5.Conclusions: PCa cells expressed high levels of CCR10, CXCR1, CXCR3 and CXCR5. Although BPH cells did not express these factors, their expression was up-regulated when BPH-1 cells were incubated with inflammatory cells. Finally, down-regulation of CXCR5 inhibited the growth and proliferation of LNCaP cells.

Labda-8(17),12,14-trien-19-oic Acid Contained in Fruits ofCupressus sempervirensSuppresses Benign Prostatic Hyperplasia in Rat andIn VitroHuman Models Through Inhibition of Androgen and STAT-3 Signaling

Fruit extract of Cupressus sempervirens (CS), which is used traditionally to treat Benign Prostatic Hyperplasia (BPH)-like urinary symptoms in patients, was scientifically validated for anti-BPH activity. The ethanolic fruit extract of CS inhibited proliferation of human BPH-stromal cells and the activity was localized to its chloroform-soluble, diterpene-rich fraction. Eight major diterpenes isolated from this fraction exhibited moderate to potent activity and the most active diterpene (labda-8(17),12,14-trien-19-oic acid) exhibited an IC50 of 37.5 μM (antiproliferative activity against human BPH-stromal cells). It significantly inhibited activation (phosphorylation) of Stat-3 in BPH-stromal cells and prevented transactivation of androgen sensitive KLK3/PSA and TMPRSS2 genes in LNCaP cells. Labda-8(17),12,14-trien-19-oic acid-rich CS fraction prevented prostatic hyperplasia in rat model and caused TUNEL labeling of stromal cells with lower expressions of IGF-I, TGF-ß and PCNA, and bcl-2/bax ratio. Human BPH tissues exhibited precise lowering of stromal component after incubation in labda-8(17),12,14-trien-19-oic acid, ex vivo. We conclude that labda-8(17),12,14-trien-19-oic acid contained in CS exhibits anti-BPH activity through inhibition of stromal proliferation and suppression of androgen action in the prostate, presenting a unique lead structure for further optimization of anti-BPH activity.

Targeting Androgen Receptor to Suppress Macrophage-induced EMT and Benign Prostatic Hyperplasia (BPH) Development

Early studies suggested macrophages might play roles in inflammation-associated benign prostatic hyperplasia (BPH) development, yet the underlying mechanisms remain unclear. Here we first showed that CD68(+) macrophages were identified in both epithelium and the stromal area of human BPH tissues. We then established an in vitro co-culture model with prostate epithelial and macrophage cell lines to study the potential impacts of infiltrating macrophages in the BPH development and found that co-culturing prostate epithelial cells with macrophages promoted migration of macrophages. In a three-dimensional culture system, the sphere diameter of BPH-1 prostate cells was significantly increased during coculture with THP-1 macrophage cells. Mechanism dissection suggested that expression levels of epithelial-mesenchymal transition (EMT) markers, such as N-cadherin, Snail, and TGF-β2, were increased, and administration of anti-TGF-β2 neutralizing antibody during co-culture suppressed the EMT and THP-1-mediated growth of BPH-1 cells, suggesting THP-1 might go through EMT to influence the BPH development and progression. Importantly, we found that modulation of androgen receptor (AR) in BPH-1 and mPrE cells significantly increased THP-1 and RAW264.7 cell migration, respectively, and enhanced expression levels of EMT markers, suggesting that AR in prostate epithelial cells might play a role in promoting macrophage-mediated EMT in prostate epithelial cells. Silencing AR function via an AR degradation enhancer, ASC-J9, decreased the macrophage migration to BPH-1 cells and suppressed EMT marker expression. Together, these results provide the first evidence to demonstrate that prostate epithelial AR function is important for macrophage-mediated EMT and proliferation of prostate epithelial cells, which represents a previously unrecognized role of AR in the cross-talk between macrophages and prostate epithelial cells. These results may provide new insights for a new therapeutic approach to battle BPH via targeting AR and AR-mediated inflammatory signaling pathways.

Increased Infiltrated Macrophages in Benign Prostatic Hyperplasia (BPH): ROLE OF STROMAL ANDROGEN RECEPTOR IN MACROPHAGE-INDUCED PROSTATE STROMAL CELL PROLIFERATION

Infiltrated macrophages may play important roles in the development and progression of benign prostatic hyperplasia (BPH), but the underlying mechanisms remain largely unknown. We found increased macrophages infiltration in human and mouse BPH tissues. By establishing a co-culture transwell system, we found increased migration of macrophages and proliferation of prostate stromal cells during co-culture. Importantly, stromal androgen receptor (AR) could enhance the migration of macrophages and macrophage-mediated stromal cell proliferation. We identified CCL3 as an AR downstream player, and found CCL3 levels were notably increased in human and mouse BPH prostates. Ablation of prostate stromal AR in a mouse BPH model significantly reduced CCL3 expression levels in prostates. Consistently, targeting AR via an AR degradation enhancer, ASC-J9®, or neutralization of CCL3 with an antibody, resulted in suppression of macrophage migration and prostate stromal cell growth. Our study provides mechanistic insights on the regulation of prostate stromal cells by macrophages via stromal AR/CCL3 signaling pathways, which could potentially allow the development of therapeutic approaches for battling BPH with persistent inflammation.

1570 TADALAFIL, THE PHOSPHODIESTERASE-5 INHIBITOR, ATTENUATES THE CELL GROWTH OF HUMAN BENIGN PROSTATIC HYPERPLASIA XENOGRAFTS IN SUPER-SCID MICE

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research1 Apr 20121570 TADALAFIL, THE PHOSPHODIESTERASE-5 INHIBITOR, ATTENUATES THE CELL GROWTH OF HUMAN BENIGN PROSTATIC HYPERPLASIA XENOGRAFTS IN SUPER-SCID MICE Hiroshi Kiuchi, Akira Tsujimura, Hidenobu Okuda, Keisuke Yamamoto, Shinichirou Fukuhara, Iwao Yoshioka, Tetsuya Takao, Yasushi Miyagawa, Norio Nonomura, and Taisei Nomura Hiroshi KiuchiHiroshi Kiuchi Suita, Japan More articles by this author , Akira TsujimuraAkira Tsujimura Suita, Japan More articles by this author , Hidenobu OkudaHidenobu Okuda Suita, Japan More articles by this author , Keisuke YamamotoKeisuke Yamamoto Suita, Japan More articles by this author , Shinichirou FukuharaShinichirou Fukuhara Suita, Japan More articles by this author , Iwao YoshiokaIwao Yoshioka Suita, Japan More articles by this author , Tetsuya TakaoTetsuya Takao Suita, Japan More articles by this author , Yasushi MiyagawaYasushi Miyagawa Suita, Japan More articles by this author , Norio NonomuraNorio Nonomura Suita, Japan More articles by this author , and Taisei NomuraTaisei Nomura Ibaraki, Japan More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.1342AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Benign prostatic hyperplasia (BPH) is characterized by a pathologic proliferation of the prostate glandular and stromal tissues, and subsequent induction of lower urinary tract symptoms. Emerging evidences suggest that phosphodiesterase-5 (PDE-5) inhibitors improve urinary symptoms in patients with BPH. However, inhibition of prostate growth by PDE-5 inhibitors has not been fully elucidated. The objective of this study is to evaluate the effect of Tadalafil, the PDE-5 inhibitor, on suppression of prostate cell growth using a novel human BPH xenograft model. METHODS Human BPH tissue was obtained from a single patient who had undertaken retropubic prostatectomy for the treatment of BPH and was transplanted under the skin of super-SCID mice; SCID mice with macrophage dysfunction. BPH tissues were collected after 2 months of treatment with Tadalafil orally or nothing (control) with sustained release testosterone pellet. Immunohistochemical study was performed to examine the histology and expression of cell type-specific markers in xenografts, including PSA, androgen receptor (AR), p63 (basal cell marker), cytokeratin 8(K8, luminal cell marker) and human-specific CD31 (human vessel marker). We next assess the effect of Tadalafil on the prostate cell proliferation by Ki67 and apoptosis by cleaved caspase-3. Endothelial NOS (eNOS) and COX-2 expression were also examined. RESULTS The implanted tissue contained normal prostatic components; the gland showed two layers of epithelial cells. Inner layer expressed K8, AR and PSA, and outer layer was positive for p63. A considerable amount of the human originated vessels was apparent. After 2 months of Tadalafil treatment, a proliferation index (percentage of Ki-positive cells) was significantly reduced, compared with control in epithelial cells (1.4% vs. 3.9%, p<0.05) as well as in stromal tissues (51.7 vs.14.1 cells/mm2, p<0.05). Cleaved caspase-3 positive cells were significantly increased by Tadalafil. In addition, Tadalafil induced up-regulation of eNOS expression and down-regulation of COX-2 expression. CONCLUSIONS Our novel xenograft model has maintained appropriate histology and protein expression, and may serve as a useful human BPH model. Our study also indicates that Tadalafil attenuates the cell growth and induces apoptosis in benign prostate cells through NO signaling. These results provide a therapeutic potential of Tadalafil for reducing prostate volume in patients with BPH. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e636 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hiroshi Kiuchi Suita, Japan More articles by this author Akira Tsujimura Suita, Japan More articles by this author Hidenobu Okuda Suita, Japan More articles by this author Keisuke Yamamoto Suita, Japan More articles by this author Shinichirou Fukuhara Suita, Japan More articles by this author Iwao Yoshioka Suita, Japan More articles by this author Tetsuya Takao Suita, Japan More articles by this author Yasushi Miyagawa Suita, Japan More articles by this author Norio Nonomura Suita, Japan More articles by this author Taisei Nomura Ibaraki, Japan More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

A cyclooxygenase-2 inhibitor reduces serum prostatic-specific antigen levels in men with benign prostatic hyperplasia and lower urinary tract symptoms and helps differentiate chronic inflammation from prostate cancer

Emerging evidence indicates that prostatic inflammation may contribute to prostatic growth in benign prostatic hyperplasia (BPH) and neoplastic changes (prostate cancer). The Medical Therapy of Prostatic Symptoms study showed that men with inflammation had a significantly higher risk of BPH progression and acute urinary retention. Evidence also shows that a cyclooxygenase-2 (COX-2) inhibitor can increase apoptotic activity in human BPH tissue. COX-2 inhibitors in combination with alpha-adrenergic blockers may increase the effectiveness of therapy for lower urinary tract symptoms (LUTS) secondary to BPH without significant side effects. COX-2 inhibitors may decrease the serum prostatic-specific antigen level and act as a biomarker to differentiate chronic inflammation from prostate cancer in patients with LUTS suggestive of BPH who do not have a palpable prostate nodule but have a serum prostatic-specific antigen level higher than 4 ng/mL. The results might provide a simple way to initially differentiate chronic inflammation from prostatic cancer. If chronic inflammation of the prostate can be reduced by COX-2 inhibitor treatment, bothersome LUTS as well as voiding conditions might be improved.

Abstract B46: The effect of sulforaphane on protein expression in benign prostatic hyperplasia tissue

Abstract Background: Benign prostatic hyperplasia (BPH) is a common condition that occurs in over 25% of men aged 50+, increasing to 50% amongst those over 80. Although the etiology of the condition has not been ascertained, the majority of BPH specimens contain inflammatory infiltrates indicating inflammation plays an important role in the progression of the disease. Furthermore, a role for inflammation has also been suggested in prostate cancer (PCa). BPH is not a pre-cursor of PCa but using BPH as a model of inflammation could result in findings that may apply and add to our understanding of PCa. Previous work has demonstrated that the consumption of broccoli can alter inflammatory pathways in the human prostate, and this may be due to the action of the isothiocyanate sulforaphane (SF), a phytochemical derived from broccoli. This dietary compound has a range of biological activities, including reducing inflammation. Hypothesis: Sulforaphane reduces inflammatory pathways and beneficially alters protein expression in BPH tissue. Methods: We obtained and cultured human BPH tissue for 24 hours with or without 10µM SF and collected tissue and media for further analysis using two distinct approaches. Firstly, in a targeted approach, known inflammatory markers were measured in the media of five tissue samples by commercially available ELISA. Secondly, a global proteomic analysis was carried out to give an unbiased view of the effect of SF on the protein expression within the tissue. Initially, protein was extracted from the tissue of three individuals, run on 2D gels and spots of interest excised from the gel and in-gel trypsin digested. The resulting peptides were analyzed on an LTQ-Orbitrap Mass Spectrometer and the proteins identified by Mascot. Selected proteins were then further quantified in additional tissue samples. Results: Analysis of the effect of SF on inflammatory markers revealed that SF significantly reduced levels of secreted IL-6 and to a lesser extent IL-8 and FGF-2. Global proteomic analysis demonstrated that there was, as expected, considerable variation between individuals, and that SF altered expression of several proteins in BPH tissue. Reduction of GRP94, an endoplasmic reticulum chaperone protein observed in one individual, was further verified in independent BPH samples. Initial immunohistochemistry in sections of BPH tissue suggests that SF reduces the nuclear localization of this protein. Further experiments are currently ongoing to investigate these effects in more individuals and also to look at the biological consequences. Conclusion: Natural variation present within humans highlights the need to use a larger number of samples in order to understand the complex effects of SF on prostate tissue. The results suggest that SF can reduce inflammatory markers and alter proteins within the human prostate indicating a possible protective role for SF in pathogenesis of the prostate. Citation Information: Cancer Prev Res 2010;3(12 Suppl):B46.

ChemInform Abstract: New Nonsteroidal Steroid 5α-Reductase Inhibitors. Syntheses and Structure-Activity Studies on Carboxamide Phenylalkyl-Substituted Pyridones and Piperidones.

In the search for nonsteroidal inhibitors of 5 alpha-reductase for the treatment of benign prostatic hyperplasia (BPH), we synthesized diisopropyl (1a-8a) and tert-butyl (1b-8b) benzamides, as well as ethyl benzoates (1c, 3c), which were substituted in 4 position via variable alkyl spacer (n = 0: 1-4, n = 1: 5, 7 and n = 3: 6, 8) with a 1-methyl-2-pyridone (1, 2, 5, 6) or a 1-methyl-2-piperidone (3, 4, 7, 8) moiety mimicking steroidal ring A. The directly connected benzamides (1a-4a, 1b-4b) and benzoates (1c, 3c) were obtained by palladium-catalysed coupling reaction of diethyl(3-pyridyl)-borane with 4-bromobenzoic acid derivatives, followed by alpha-oxidation of the 1-methyl-pyridinium salt and subsequent separation of the regioisomers. Catalytic hydrogenation of the pyridones (1, 2) led to the piperidones (3, 4). The preparation of the benzamides with a methylene (5, 7) and a propylene spacer (6, 8), respectively, started with the reduction of the keto group of 5-benzoyl-1,2-dihydro-1-methyl-2(1H)-pyridone and catalytic hydrogenation of the alkene obtained by Wittig reaction of 5-formyl-1,2-dihydro-1-methyl-2(1H)-pyridone with (2-phenylethyl)triphenylphosphonium bromide, respectively. The phenyl ring was functionalized by Friedel-Crafts reaction, haloform cleavage to give the acid, formation of the acid chloride, and subsequent treatment with the appropriate amines. Again, catalytic hydrogenation of the pyridones (5, 6) led to the piperidones (7, 8). The 5 alpha-reductase inhibitory properties were determined using rat ventral prostate, as well as human BPH tissue as enzyme source, 1 beta-2 beta-[3H]testosterone as substrate and a HPLC procedure for the separation of dihydrotestosterone (DHT). Tested at a concentration of 100 microM, the inhibition values of 1-8 ranged from 0-79%. Significant differences were observed between rat and human enzyme. The most active compound was ethyl 4-(1-methyl-2-oxopiperid-5-yl)benzoate 3c (68%) for the human enzyme and N,N-bis(1-methylethyl)-4-[3-(1,2-dihydro-1-methyl-2-oxopyrid-5-yl) propyl] benzamide 6a (79%) for the rat enzyme.