Growth hormone-releasing hormone (GHRH) is secreted by the hypothalamus and acts on the pituitary gland to stimulate the release of growth hormone (GH). GHRH can also be produced by human cancers, in which it functions as an autocrine/paracrine growth factor. We have previously shown that synthetic antagonistic analogues of GHRH are able to successfully suppress the growth of 60 different human cancer cell lines representing over 20 cancers. Nevertheless, the expression of GHRH and its receptors in leukaemias has never been examined. Our study demonstrates the presence of GHRH receptor (GHRH-R) on 3 of 4 human acute myeloid leukaemia (AML) cell lines-K-562, THP-1, and KG-1a-and significant inhibition of proliferation of these three cell lines in vitro following incubation with the GHRH antagonist MIA-602. We further show that this inhibition of proliferation is associated with the upregulation of pro-apoptotic genes and inhibition of Akt signalling in leukaemic cells. Treatment with MIA-602 of mice bearing xenografts of these human AML cell lines drastically reduced tumour growth. The expression of GHRH-R was further confirmed in 9 of 9 samples from patients with AML. These findings offer a new therapeutic approach to this malignancy and suggest a possible role of GHRH-R signalling in the pathology of AML.

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2018MP45-05 GROWTH HORMONE-RELEASING HORMONE (GHRH) ANTAGONISTS INHIBIT PROSTATIC ENLARGEMENT AND INFLAMMATION IN ?-CARRAGEENAN-INDUCED EXPERIMENTAL PROSTATITIS Petra Popovics, Ferenc G. Rick, Rhenzi Cai, Wei Sha, and Andrew V. Schally Petra PopovicsPetra Popovics More articles by this author , Ferenc G. RickFerenc G. Rick More articles by this author , Rhenzi CaiRhenzi Cai More articles by this author , Wei ShaWei Sha More articles by this author , and Andrew V. SchallyAndrew V. Schally More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.1444AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The pathogenesis of benign prostatic hyperplasia (BPH) is associated with chronic inflammatory processes, but the complex molecular mechanism is not entirely explored. Our previous studies demonstrated that prostatic levels of GHRH are increased in experimental models of prostate cancer, BPH, and autoimmune prostatitis whereas GHRH antagonists exert multiple anti-proliferative and anti-inflammatory effects in these tissues. Our current study tests whether GHRH antagonists confer beneficial effects on chronic prostatic inflammation in rats. METHODS Chronic inflammation was induced in Sprague-Dawley rats by two intraprostatic injections of 3% λ-carrageenan into both right and left ventral lobes, three weeks apart. GHRH antagonist, MIA-690, was administered 5 days after the second intraprostatic injection at 20 μg daily dose for 4 weeks. Prostates were processed for histology, immunohistochemistry, western blot analyses and quantitative PCR. In vitro effects of GHRH were tested on human prostate epithelial (BPH-1) and stromal (WPMY-1) cell lines. RESULTS Carrageenan-induced prostatitis (CIP) produced a 73.2% increase in weights of the ventral prostate lobes which was reduced by 19.2% in rats treated with MIA-690. Vimentin staining showed thickening of the stromal compartment which effect was also suppressed by GHRH antagonists. Western blot analysis demonstrated an increase in GHRH, COX2 and TGF-β1 levels by inflammation and suppression by MIA-690. Prostatic IGF-1 levels measured by ELISA were increased by 65% in CIP and were suppressed to control level by GHRH antagonist. A qPCR array analysis of genes related to EMT revealed upregulation in several genes including collagens, matrix metalloproteinase 9, Snail1, TGF-β1 and vimentin by inflammation and downregulation by treatment with MIA-690. In vitro, GHRH stimulated the phosphorylation of EGF and IGF receptors and upregulated the level of TGF-β1 in BPH-1 cells but had no measurable effects on WPMY-1 stromal cells. CONCLUSIONS Our current findings strongly indicate that GHRH is a key molecular player in inflammation-induced prostate enlargement via its direct action on epithelial cells. The effect of GHRH antagonists on stromal cells may be the consequence of their effect on epithelial growth factor production. Consequently, GHRH antagonists could be clinically useful to treat early and advanced stages of BPH due to their anti-proliferative and anti-inflammatory activities. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e598-e599 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Petra Popovics More articles by this author Ferenc G. Rick More articles by this author Rhenzi Cai More articles by this author Wei Sha More articles by this author Andrew V. Schally More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

BackgroundInflammation plays a key role in the etiology of benign prostatic hyperplasia (BPH) through multiple pathways involving the stimulation of proliferation by cytokines and growth factors as well as the induction of the focal occurrence of epithelial‐to‐mesenchymal transition (EMT). We have previously reported that GHRH acts as a prostatic growth factor in experimental BPH and in autoimmune prostatitis models and its blockade with GHRH antagonists offer therapeutic approaches for these conditions. Our current study was aimed at the investigation of the beneficial effects of GHRH antagonists in λ‐carrageenan‐induced chronic prostatitis and at probing the downstream molecular pathways that are implicated in GHRH signaling.MethodsTo demonstrate the complications triggered by recurrent/chronic prostatic inflammation in Sprague‐Dawley rats, 50 μL 3% carrageenan was injected into both ventral prostate lobes two times, 3 weeks apart. GHRH antagonist, MIA‐690, was administered 5 days after the second intraprostatic injection at 20 μg daily dose for 4 weeks. GHRH‐induced signaling events were identified in BPH‐1 and in primary prostate epithelial (PrEp) cells at 5, 15, 30, and 60 min with Western blot.ResultsInflammation induced prostatic enlargement and increased the area of the stromal compartment whereas treatment with the GHRH antagonist significantly reduced these effects. This beneficial activity was consistent with a decrease in prostatic GHRH, inflammatory marker COX‐2, growth factor IGF‐1 and inflammatory and EMT marker TGF‐β1 protein levels and the expression of multiple genes related to EMT. In vitro, GHRH stimulated multiple pathways involved in inflammation and growth in both BPH‐1 and PrEp cells including NFκB p65, AKT, ERK1/2, EGFR, STAT3 and increased the levels of TGF‐β1 and Snail/Slug. Most interestingly, GHRH also stimulated the transactivation of the IGF receptor.ConclusionsThe study demonstrates that GHRH antagonists could be beneficial for the treatment of prostatic inflammation and BPH in part by inhibiting the growth‐promoting and inflammatory effects of locally produced GHRH.

The syntheses and biological evaluations of new GHRH analogs of Miami (MIA) series with greatly increased anticancer activity are described. In the design and synthesis of these analogs, the following previous substitutions were conserved: D-Arg(2), Har(9), Abuls(15), and Nle(27). Most new analogs had Ala at position 8. Since replacements of both Lys(12) and Lys(21) with Orn increased resistance against enzymatic degradation, these modifications were kept. The substitutions of Arg at both positions 11 and 20 by His were also conserved. We kept D-Arg(28), Har(29) -NH2 at the C-terminus or inserted Agm or 12-amino dodecanoic acid amide at position 30. We incorporated pentafluoro-Phe (Fpa(s)), instead of Cpa, at position 6 and Tyr(Me) at position 10 and omega-amino acids at N-terminus of some analogs. These GHRH analogs were prepared by solid-phase methodology and purified by HPLC. The evaluation of the activity of the analogs on GH release was carried out in vitro on rat pituitaries and in vivo in male rats. Receptor binding affinities were measured in vitro by the competitive binding analysis. The inhibitory activity of the analogs on tumor proliferation in vitro was tested in several human cancer cell lines such as HEC-1A endometrial adenocarcinoma, HCT-15 colorectal adenocarcinoma, and LNCaP prostatic carcinoma. For in vivo tests, various cell lines including PC-3 prostate cancer, HEC-1A endometrial adenocarcinoma, HT diffuse mixed beta cell lymphoma, and ACHN renal cell carcinoma cell lines were xenografted into nude mice and treated subcutaneously with GHRH antagonists at doses of 1-5 mu g/day. Analogs MIA-602, MIA-604, MIA-610, and MIA-640 showed the highest binding affinities, 30, 58, 48, and 73 times higher respectively, than GHRH (1-29) NH2. Treatment of LNCaP and HCT-15 cells with 5 mu M MIA-602 or MIA-690 decreased proliferation by 40%-80%. In accord with previous tests in various human cancer lines, analog MIA-602 showed high inhibitory activity in vivo on growth of PC-3 prostate cancer, HT-mixed beta cell lymphoma, HEC-1A endometrial adenocarcinoma and ACHN renal cell carcinoma. Thus, GHRH analogs of the Miami series powerfully suppress tumor growth, but have only a weak endocrine GH inhibitory activity. The suppression of tumor growth could be induced in part by the downregulation of GHRH receptors levels. Published by Elsevier Inc.

Significance The current therapeutic approaches to the treatment of benign prostatic hyperplasia (BPH) do not take into consideration that inflammation is an important factor in the pathogenesis of this disease. We previously demonstrated that growth hormone-releasing hormone (GHRH) antagonists reduce prostatic weights and decrease the level of inflammatory cytokines in a testosterone-induced BPH model. This study sheds light on the paracrine roles of GHRH in prostatic inflammation and demonstrates that GHRH stimulates the growth of BPH-1 and primary prostate epithelial spheres and that GHRH antagonists reduce prostate volume in an experimental model of prostatic inflammation.

The discovery, isolation, elucidation of structure, synthesis, and initial testing of the neuropeptide hypothalamic luteinizing hormone‐releasing hormone (LHRH), which regulates reproduction, is briefly described. The design, synthesis, and experimental and clinical testing of agonistic analogs of LHRH is extensively reviewed focusing on the development of new methods for the treatment of prostate cancer. Subsequent development of antagonistic analogs of LHRH is then faithfully recounted with special emphasis on therapy of prostate cancer and BPH. The concepts of targeted therapy to peptide receptors on tumors are re‐examined and the development of the cytotoxic analogs of LHRH and their status is reviewed. The endeavor to develop better therapies for prostate cancer, based on LHRH analogs, guided much of our work.

PURPOSE:To explore how follicle-stimulating hormone (FSH) may contribute to cardiovascular, metabolic, skeletal, and cognitive events in men treated for prostate cancer, with various forms of androgen deprivation therapy (ADT). MATERIALS AND METHODS:A colloquium of prostate cancer experts was convened in May 2015, to discuss the role of FSH in the development of unwanted effects associated with ADT. Subsequently, a literature review (Medline, PubMed, and relevant congress abstract databases) was performed to further explore and evaluate the collected evidence. RESULTS:It has become evident that, in the setting of ADT, FSH can promote the development of atherosclerotic plaque formation, metabolic syndrome, and insulin resistance. Data also suggest that FSH is an important mediator of bone remodeling, particularly bone resorption, and thereby increases the risk for bone fracture. Additional evidence implicates a role for FSH in bone metastasis as well. The influence of FSH on ADT-induced cognitive deficits awaits further elucidation; however, the possibility that FSH may be involved therein cannot be ruled out. CONCLUSIONS:The widespread molecular and physiological consequences of FSH system activation in normal and pathological conditions are becoming better understood. Progress in this area has been achieved by the development of additional investigative and clinical measures to better evaluate specific adverse effects. More research is needed on FSH function in the development of cancer as well as its association with cardiovascular, metabolic, musculoskeletal, and cognitive effects in ADT.

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2017MP17-03 INFLAMMATION-INDUCED PROSTATIC ENLARGEMENT AND PROLIFERATION OF PROSTATE EPITHELIAL CELLS IS REDUCED BY GROWTH HORMONE-RELEASING HORMONE (GHRH) ANTAGONISTS THROUGH THE INHIBITION OF EPITHELIAL-TO-MESENCHYMAL TRANSITION Petra Popovics, Andrew Schally, Luis Salgueiro, Krisztina Kovacs, and Ferenc Rick Petra PopovicsPetra Popovics More articles by this author , Andrew SchallyAndrew Schally More articles by this author , Luis SalgueiroLuis Salgueiro More articles by this author , Krisztina KovacsKrisztina Kovacs More articles by this author , and Ferenc RickFerenc Rick More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.592AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The pathogenesis of benign prostatic hyperplasia (BPH) has been associated with various factors including hormonal imbalance, inflammation-induced cell proliferation and epithelial-to-mesenchymal transition. We have previously demonstrated that prostatic GHRH and its receptors are upregulated in a rat testosterone-induced BPH model and GHRH antagonists suppress the levels of proinflammatory cytokines. Based on these findings, we investigated the role of GHRH in inflammation-induced proliferation of prostate epithelial cells in vitro and prostate enlargement in experimental autoimmune prostatitis. METHODS Autoimmune inflammation in the prostates of Balb/c mice was induced by subcutaneous injections of rat male tissue homogenate. Changes in prostate volume were measured with the VEVO® 1100 ultrasound imaging system. Human BPH-1 and primary prostate epithelial cells were used in matrigel-embedded 3D cultures and average sphere diameters were evaluated. Chronic inflammation was mimicked by treating cells with THP-1 macrophage-conditioned medium or Il-17A, whereas EMT was triggered with TGF-β1 or TGF-β2 peptides. The role of secreted GHRH in inflammation-induced proliferation was determined by using GHRH antagonists developed in our lab. Changes in the protein levels were determined by western blot. RESULTS Experimental autoimmune prostatitis increased the volume of the ventral prostate by 92% at week 8 compared to control (p<0.001). A 1-month daily treatment with GHRH antagonists caused a significant, 48% reduction in prostate volume. Macrophage-conditioned medium induced a 26% increase (p<0.001) in the average diameter of cells and elevation in the expression of mesenchymal markers. The mRNA and protein expression of GHRH were significantly increased by macrophage-conditioned medium. GHRH antagonist reduced inflammation- and TGF-β2-induced increase in diameter by 64% (p<0.01) and by 67% (p<0.001), respectively, and reduced the expression of n-cadherin. The stimulatory effect of TGF-β2 was abolished when GHRH receptor expression was silenced by stable transfection of shRNA. IL-17A stimulation of the growth of primary epithelial cells were also significantly reduced by GHRH antagonists. CONCLUSIONS Our results indicate that GHRH is a key factor in prostatic inflammation-induced prostate enlargement and EMT and suggest that GHRH antagonists have beneficial effects in chronic prostatitis and BPH. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e212 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Petra Popovics More articles by this author Andrew Schally More articles by this author Luis Salgueiro More articles by this author Krisztina Kovacs More articles by this author Ferenc Rick More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Androgen deprivation therapy (ADT) of androgen-dependent prostate cancer (PCa), which is still the gold standard treatment, was based Huggins and Hodges' 1941 finding that the growth of PCa cells requires the androgen, testosterone (Rick and Schally, 2015Rick F.G. Schally A.V. Bench-to-bedside development of agonists and antagonists of luteinizing hormone-releasing hormone for treatment of advanced prostate cancer.Urol. Oncol. 2015; 33: 270-274Summary Full Text Full Text PDF PubMed Scopus (26) Google Scholar). In addition, an increasing body of evidence has revealed that the development and progression of androgen-dependent PCa to castration-resistant prostate cancer (CRPC) have an intimate association with the androgen–androgen receptor axis (AR axis) (Knudsen and Scher, 2009Knudsen K.E. Scher H.I. Starving the addiction: new opportunities for durable suppression of AR signaling in prostate cancer.Clin. Cancer Res. 2009; 15: 4792-4798Crossref PubMed Scopus (249) Google Scholar). The application of AR axis drugs, such as the 5α-reductase inhibitors (5ARI), finasteride and dutasteride, which decrease levels of dihydrotestosterone (DHT) in order to prevent development or progression of PCa, remains to be extensively discussed and is controversial (Kosaka et al., 2014Kosaka T. Miyajima A. Oya M. Is DHT production by 5alpha-reductase friend or foe in prostate cancer?.Front. Oncol. 2014; 4: 247Crossref PubMed Scopus (9) Google Scholar). The concept is driven by the results of two large, randomized, placebo-controlled trials: the Prostate Cancer Prevention Trial (PCPT) with finasteride (Thompson et al., 2003Thompson I.M. Goodman P.J. Tangen C.M. et al.The influence of finasteride on the development of prostate cancer.N. Engl. J. Med. 2003; 349: 215-224Crossref PubMed Scopus (2284) Google Scholar) and the Reduction by Dutasteride of Prostate Cancer Events (REDUCE) trial (Andriole et al., 2010Andriole G.L. Bostwick D.G. Brawley O.W. et al.Effect of dutasteride on the risk of prostate cancer.N. Engl. J. Med. 2010; 362: 1192-1202Crossref PubMed Scopus (900) Google Scholar). In the PCPT, finasteride significantly reduced the overall risk of prostate cancer but cancers with high Gleason scores (7–10) were found in 6.4% of the tumors in the finasteride group, compared with only 5.1% of those being found in the placebo group. In a manner analogous to the PCPT trial, the REDUCE trial reported an overall reduction in the number of PCa patients (with a low Gleason score of 5–6) in those receiving dutasteride versus those given a placebo (19.9% and 25.1%, respectively); similarly, tumors with a high Gleason score (8–10) were more frequent in the dutasteride-treated group than in the placebo group. In the Reduction by Dutasteride of Clinical Progression Events in Expectant Management (REDEEM) trial, dutasteride was associated with a 38% decrease in the cancer detection rate on repeat biopsy at year 3 in men with low-grade Gleason score (5–6) prostate cancer undergoing active surveillance and who received three years of treatment with dutasteride or placebo (Fleshner et al., 2012Fleshner N.E. Lucia M.S. Egerdie B. et al.Dutasteride in localised prostate cancer management: the REDEEM randomised, double-blind, placebo-controlled trial.Lancet. 2012; 379: 1103-1111Summary Full Text Full Text PDF PubMed Scopus (159) Google Scholar). Because of concerns regarding the possible induction of de novo aggressive PCa, the US Food and Drug Administration did not grant approval of 5ARIs for the chemoprevention of PCa in December 2010 (Theoret et al., 2011Theoret M.R. Ning Y.M. Zhang J.J. Justice R. Keegan P. Pazdur R. The risks and benefits of 5alpha-reductase inhibitors for prostate-cancer prevention.N. Engl. J. Med. 2011; 365: 97-99Crossref PubMed Scopus (130) Google Scholar). However, it is still unclear whether the observed increase in high-grade CaP in these trials was real or artifact (Lucia et al., 2007Lucia M.S. Epstein J.I. Goodman P.J. et al.Finasteride and high-grade prostate cancer in the prostate cancer prevention trial.J. Natl. Cancer Inst. 2007; 99: 1375-1383Crossref PubMed Scopus (206) Google Scholar). These observations cannot be fully explained from a purely mechanistic point of view. Therefore, further basic and clinical investigations are necessary (Kosaka et al., 2014Kosaka T. Miyajima A. Oya M. Is DHT production by 5alpha-reductase friend or foe in prostate cancer?.Front. Oncol. 2014; 4: 247Crossref PubMed Scopus (9) Google Scholar). In this issue of EBioMedicine, Kim et al. seek to improve biologic understanding of the grade-specific effects of the 5ARI, finasteride, by studying 183 men with localized prostate cancer, who were randomized to receive 5 mg of finasteride or placebo daily for 4 to 6 weeks pre-prostatectomy (Kim et al., 2016Kim J. Davis J.W. Klein E.A. et al.Tissue effects in a randomized controlled trial of short-term finasteride in early prostate cancer.EBioMedicine. 2016; 7: 85-93Summary Full Text Full Text PDF PubMed Scopus (5) Google Scholar). In fact, this is one of the few studies done in early prostate cancer to investigate the time it takes for changes in gene expression to occur following finasteride therapy. The primary end point was to compare the frequency of expression of a predefined high-grade molecular signature (ERβ, UBE2C, SRD5A2, and VEGF) differentiating high- and low-grade tumors in the Gleason grade (GG) 3 areas of finasteride-exposed tumors with those of placebo-exposed tumors, adjusted for Gleason score (GS) at prostatectomy. Secondary endpoints included assessment of androgen receptor (AR) levels, Ki-67, and cleaved caspase 3 to estimate the effects of finasteride on the expression of its downstream targets, cell proliferation, and apoptosis, respectively. Unfortunately, the primary endpoint could not be assessed as the predetermined molecular signature was not able to distinguish GG3 from GG4 areas in the placebo group. However, expression of AR was significantly lower in the GG4 areas of the finasteride group compared to those of the placebo group (Kim et al., 2016Kim J. Davis J.W. Klein E.A. et al.Tissue effects in a randomized controlled trial of short-term finasteride in early prostate cancer.EBioMedicine. 2016; 7: 85-93Summary Full Text Full Text PDF PubMed Scopus (5) Google Scholar). The authors claim that this finding is in accord with an emerging concept that reduced androgens in prostate tissues may, over time, lead to de-repression of AR expression, which, in turn, deregulates AR function and downstream de-repression of the AR target genes normally suppressed by androgens (Kim et al., 2016Kim J. Davis J.W. Klein E.A. et al.Tissue effects in a randomized controlled trial of short-term finasteride in early prostate cancer.EBioMedicine. 2016; 7: 85-93Summary Full Text Full Text PDF PubMed Scopus (5) Google Scholar). Within the finasteride group, AR expression was also lower in GG4 than in GG3 areas, but not significantly. Expression of the apoptosis marker, cleaved caspase 3, in GG3 and GG4 tumor areas was significantly increased after short-term exposure to finasteride, consistent with preventive efficacy, as shown in the PCPT, and was lower in GG4 than in GG3 areas within both treatment and placebo groups (Kim et al., 2016Kim J. Davis J.W. Klein E.A. et al.Tissue effects in a randomized controlled trial of short-term finasteride in early prostate cancer.EBioMedicine. 2016; 7: 85-93Summary Full Text Full Text PDF PubMed Scopus (5) Google Scholar). In the literature, conflicting findings on the influence of 5ARIs have been reported, nonetheless, it seems that the molecular effects of 5ARIs depend on the exposure duration. One major limitation of this trial is the short period over which the study was conducted: molecular alterations in tumors exposed to finasteride may occur over a longer period of time than 4–6 weeks. Another possible explanation is that the study was underpowered: the original projections of 100 patients for each group may have been inadequate. In addition, the evaluation of tumor samples was restricted to the peripheral zone and to samples with GG patterns primarily presenting poorly formed and fused glands. It is not known what changes occur in tumors of transition zone origin or other GG4 patterns. In summary, this overall well-designed trial by Kim et al. emphasizes the necessity for further investigation of time-based effects of finasteride on molecular changes and their basic and clinical importance. Confirmation and extension of these findings may result in a valuable test allowing the identification of finasteride-responsive tumors in order to provide personalized care and/or in improved estimates of the risk of progression of the individual's disease. The authors declared no conflicts of interest relevant to this manuscript. Tissue Effects in a Randomized Controlled Trial of Short-term Finasteride in Early Prostate CancerWe showed that finasteride's effect on apoptosis and AR expression is tumor grade dependent after short-term intervention. This may explain finasteride's selective suppression of low-grade tumors observed in the PCPT. Full-Text PDF Open AccessAlcohol and Steatosis: The Japanese ParadoxIn this issue of EBioMedicine, Roerecke and Colleagues report that, in Japan, alcohol consumption as low as <20 g daily was associated with significant protection from incident and prevalent fatty liver; however, no such association was found in countries other than Japan (Roerecke et al., 2016). This systematic review is based on the analysis of 18 articles (11 of which are from Japan) which recruited, overall, 99,370 participants, 25,662 of whom had steatosis. Full-Text PDF Open AccessAlcohol and steatosis: The Japanese paradox — Authors' replyWe thank Lonardo et al. (2016) for their comments on our meta-analysis on the relationship between alcohol consumption and hepatic steatosis (Roerecke et al., 2016). We would like to clarify our position on overall risk associated with alcohol consumption and specifically on the risk of hepatic steatosis. First, as we have stated in our paper, we are in agreement with Lonardo et al. that a distinction between alcoholic and non-alcoholic hepatic steatosis has little meaning because alcohol is only one of the risk factors for liver disease, including non-alcoholic fatty liver disease. Full-Text PDF Open Access

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology1 Apr 2016MP44-11 GROWTH HORMONE-RELEASING HORMONE (GHRH) ANTAGONISTS REDUCE INFLAMMATION- AND TRANSFORMING GROWTH FACTOR (TGF)-?2-INDUCED PROLIFERATION OF HUMAN BPH-1 PROSTATE EPITHELIAL CELLS GROWN IN 3D CULTURE Petra Popovics, Andrew V. Schally, Roberto Perez, Rhenzi Cai, and Ferenc G. Rick Petra PopovicsPetra Popovics More articles by this author , Andrew V. SchallyAndrew V. Schally More articles by this author , Roberto PerezRoberto Perez More articles by this author , Rhenzi CaiRhenzi Cai More articles by this author , and Ferenc G. RickFerenc G. Rick More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.267AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Benign prostatic hyperplasia (BPH) is frequently associated with chronic inflammation. Epithelial-to-mesenchymal transition (EMT) has been recently indicated as a major factor in the pathogenesis of BPH. Evidence also suggests that inflammatory cytokines may induce BPH. Hypothalamic neuropeptides, GHRH, luteinizing hormone-releasing hormone (LHRH) and their receptors are expressed in experimental models of BPH in which antagonists of GHRH and LHRH have been shown to suppress the levels of proinflammatory cytokines and alter the expression of genes related to EMT. These findings imply a role of GHRH/LHRH in the development of prostatitis, however, this has not been investigated to date. METHODS Human BPH-1 prostatic cell line was used to generate matrigel-embedded 3D cultures in which average sphere diameters were evaluated. Chronic inflammation was induced by treating cells with THP-1 macrophage-conditioned medium whereas EMT was triggered with TGF-ß1 or TGF-ß2 peptides. The role of secreted LHRH and GHRH in inflammation-induced proliferation was determined by using the LHRH antagonist, degarelix, and a specific GHRH antagonist developed in our lab. The presence of receptors for GHRH and LHRH was confirmed in 3D cultures by immunocytochemistry and quantitative RT-PCR. RESULTS BPH-1-derived spherical structures expressed cell membrane receptors for LHRH and GHRH. Macrophage-conditioned medium induced a 26% increase (P<0.001) in the average diameter of cells and a 2-fold elevation in the expression of the EMT markers N-cadherin (p<0.01) and Snail (p<0.001) as detected by qPCR. The expression of GHRH was also increased 2.7-fold (p<0.001), whereas mRNA levels of LHRH were slightly reduced (ns). GHRH antagonist reduced inflammation-induced increase in diameter by 64% (P<0.01), while no significant change was observed after degarelix treatment. TGF-ß2 increased average sphere diameter by 32%, but this effect was reduced by 67% by the GHRH antagonist (P<0.001). In contrast, TGF-ß1 did not significantly affect sphere diameter. CONCLUSIONS This study identifies GHRH as an important factor in inflammation-induced proliferation and TGF-ß2-triggered EMT of prostate epithelial cells and suggests the merit of further investigation to elucidate the effects of GHRH antagonists in the reduction of inflammation-induced BPH. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e602-e603 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Petra Popovics More articles by this author Andrew V. Schally More articles by this author Roberto Perez More articles by this author Rhenzi Cai More articles by this author Ferenc G. Rick More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Receptors for LHRH (luteinizing hormone-releasing hormone) are expressed in about 80% of human endometrial, ovarian and prostate cancers and are also found in more than 50% of breast cancers including triple negative breast cancers. In the human body, LHRH receptors are found at significant levels in the pituitary and reproductive organs. Other benign tissues or hematopoietic stem cells express only low levels of receptors for LHRH or no receptors. Thus LHRH receptors are promising targets for a receptor- mediated chemotherapy with cytotoxic hybrid molecules. Cytotoxic analogs of LHRH consist of a LHRH agonist, which is used as a carrier peptide and DOX or its derivatives. Cytotoxic analogs of LHRH, AEZS-108 (formerly known as AN-152) and AN-207, exhibit anti-cancer activity in various in vitro and in vivo models of LHRH-receptor positive cancers. In AEZS-108 (zoptarelin DOX) DOX is covalently linked to the LHRH agonist [D-Lys(6)]LHRH. Results of phase I and II clinical studies in patients with breast, endometrial and ovarian cancers demonstrated good anticancer activity with moderate toxic side effects and without any sign of cardiotoxicity so far. AEZS-108 is also being evaluated in phase I/II studies in castration resistant prostate cancer and metastatic bladder cancer. Because of the very promising phase II results in endometrial cancer, a multinational, multicenter phase III study of this malignancy has been initiated and is currently recruiting patients.

Dyslipidemia associated with triglyceride-rich lipoproteins (TRLs) represents an important residual risk factor for cardiovascular and chronic kidney disease in patients with type 1 diabetes (T1D). Levels of growth hormone (GH) are elevated in T1D, which aggravates both hyperglycemia and dyslipidemia. The hypothalamic growth hormone-releasing hormone (GHRH) regulates the release of GH by the pituitary but also exerts separate actions on peripheral GHRH receptors, the functional role of which remains elusive in T1D. In a rat model of streptozotocin (STZ)-induced T1D, GHRH receptor expression was found to be up-regulated in the distal small intestine, a tissue involved in chylomicron synthesis. Treatment of T1D rats with a GHRH antagonist, MIA-602, at a dose that did not affect plasma GH levels, significantly reduced TRL, as well as markers of renal injury, and improved endothelial-dependent vasorelaxation. Glucagon-like peptide 1 (GLP-1) reduces hyperglucagonemia and postprandial TRL, the latter in part through a decreased synthesis of apolipoprotein B-48 (ApoB-48) by intestinal cells. Although plasma GLP-1 levels were elevated in diabetic animals, this was accompanied by increased rather than reduced glucagon levels, suggesting impaired GLP-1 signaling. Treatment with MIA-602 normalized GLP-1 and glucagon to control levels in T1D rats. MIA-602 also decreased secretion of ApoB-48 from rat intestinal epithelial cells in response to oleic acid stimulation in vitro, in part through a GLP-1-dependent mechanism. Our findings support the hypothesis that antagonizing the signaling of GHRH in T1D may improve GLP-1 function in the small intestine, which, in turn, diminishes TRL and reduces renal and vascular complications.

433 Background: Urinary bladder cancer is the fifth most frequent cancer diagnosed and among the most expensive cancers to treat in the United States. The management of muscle-invasive tumors presents a clinical challenge because of the toxicity and limitations in efficacy and durability of current therapeutic modalities. Novel therapeutic strategies for this disease are of paramount importance. Growth hormone-releasing hormone (GHRH) receptors and its splice variant were detected in a series of urothelial malignancies and GHRH has been shown to influence the growth of these tumors. Herein we evaluated the effect of GHRH antagonists on the growth of various experimental human urinary bladder cancers in vitro and in vivo in nude mice. Methods: We investigated the effects of several GHRH antagonists MIA 602, MIA 606 and MIA 690 on growth of urothelial HT-1376, J82, and RT-4 tumors xenografted into nude mice. The presence of GHRH receptors was validated by Western blotting. Results: The receptors for GHRH and their main splice variant, SV1, were detected in tumor samples of all 3 human bladder cancer models. In the HT-1376 tumors, the GHRH antagonists caused a 30-60% reduction in volume and 52-70% decrease in tumor weights (p < 0.05). All three antagonists strongly inhibited growth of J82 cancers as shown by a 62-75% reduction in tumor volume and 54-66% decrease in tumor weights (p < 0.05). Treatment with MIA-602 and MIA-606 resulted in a similar marked inhibition of growth of RT-4 cancers; tumor volume and weights were reduced by about 51-71% in the treated groups (p < 0.05). The mice tolerated this therapy well; body and organ weights were not significantly changed by the treatments. No toxicity from the GHRH antagonists was noted. Conclusions: We demonstrated the efficacy of potent GHRH antagonists and their lack of toxicity in inhibiting the growth of experimental models of bladder cancer in vivo. The expression of GHRH receptors was detected in all 3 models of human primary urothelial bladder carcinomas. Our findings warrant further development of GHRH antagonists for clinical therapy of bladder cancer alone or in combination with current chemotherapeutic agents and Exploration of their mechanism of action.

It is a pleasure to contribute our presentation at the International Prostate Forum of the Annual Meeting of the American Urological Association (AUA) to this special issue of the Asian Journal of Andrology.

Growth hormone releasing hormone (GHRH) from hypothalamus nominatively stimulates growth hormone release from adenohypophysis. GHRH is also produced by cancers, acting as an autocrine/paracrine growth factor. This growth factor function is seen in lymphoma, melanoma, colorectal, liver, lung, breast, prostate, kidney, bladder cancers. Pituitary type GHRH receptors and their splice variants are also expressed in these malignancies. Synthetic antagonists of the GHRH receptor inhibit proliferation of cancers. Besides direct inhibitory effects on tumors, GHRH antagonists also enhance cytotoxic chemotherapy. GHRH antagonists potentiate docetaxel effects on growth of H460 non-small cell lung cancer (NSCLC) and MX-1 breast cancer plus suppressive action of doxorubicin on MX-1 and HCC1806 breast cancer. We investigated mechanisms of antagonists on tumor growth, inflammatory signaling, doxorubicin response, expression of drug resistance genes, and efflux pump function. Triple negative breast cancer cell xenografted into nude mice were treated with GHRH antagonist, doxorubicin, or their combination. The combination reduced tumor growth, inflammatory gene expression, drug-resistance gene expression, cancer stem-cell marker expression, and efflux-pump function. Thus, antagonists increased the efficacy of doxorubicin in HCC1806 and MX-1 tumors. Growth inhibition of H460 NSCLC by GHRH antagonists induced marked downregulation in expression of prosurvival proteins K-Ras, COX-2, and pAKT. In HT-29, HCT-116 and HCT-15 colorectal cancer lines, GHRH antagonist treatment caused cellular arrest in S-phase of cell cycle, potentiated inhibition of in vitro proliferation and in vivo growth produced by S-phase specific cytotoxic agents, 5-FU, irinotecan and cisplatin. This enhancement of cytotoxic therapy by GHRH antagonists should have clinical applications.

Background We previously showed that growth hormone-releasing hormone (GHRH) agonists are cardioprotective following myocardial infarction (MI). Here, our aim was to evaluate the in vitro and in vivo activities of highly potent new GHRH agonists, and elucidate their mechanisms of action in promoting cardiac repair. Methods and Results H9c2 cells were cultured in serum-free medium, mimicking nutritional deprivation. GHRH agonists decreased calcium influx and significantly improved cell survival. Rats with cardiac infarction were treated with GHRH agonists or placebo for four weeks. MI size was reduced by selected GHRH agonists (JI-38, MR-356, MR-409); this accompanied an increased number of cardiac c-kit+ cells, cellular mitotic divisions, and vascular density. One week post-MI, MR-409 significantly reduced plasma levels of IL-2, IL-6, IL-10 and TNF-α compared to placebo. Gene expression studies revealed favorable outcomes of MR-409 treatment partially result from inhibitory activity on pro-apoptotic molecules and pro-fibrotic systems, and by elevation of bone morphogenetic proteins. Conclusions Treatment with GHRH agonists appears to reduce the inflammatory responses post-MI and may consequently improve mechanisms of healing and cardiac remod eling by regulating pathways involved in fibrosis, apoptosis and cardiac repair. Patients with cardiac dysfunction could benefit from treatment with novel GHRH agonists.

Background: Androgen deprivation therapy (ADT) has been the standard of care for treating patients with hormone-sensitive advanced prostate cancer (PCa) for 3 decades. The agonists of luteinizing hormone releasing hormone (LBRH), also called gonadotropin-releasing hormone, are still the most frequently used form of medical ADT.ADT and LHRH analogs: The application of agonists of LHRH has improved and modernized the treatment of advanced PCa; millions of patients have benefited from therapy with LHRH agonists as a preferred alternative to surgical castration, as the psychological effects and perpetuity of orchiectomy are undesirable for most men. Despite their efficacy, agonists of LHRH have several shortcomings, including initial surge in testosterone, producing exacerbation of clinical symptoms, and microsurges in testosterone that might occur after each administration. A new, alternate approach to ADT is emerging with the improvements in antagonists of LHRH. This class of LHRH analogues produces a direct and immediate blockade of pituitary LHRH receptors and leads to a more rapid suppression of testosterone without an initial surge or subsequent microsurges. Degarelix, a third-generation LHRH antagonist, is the only antagonist with a low histamine-releasing activity that is currently on the market for clinical use in advanced PCa with improved testosterone suppression, better control of follicle-stimulating hormone and prostate-specific antigen, and which offers a prolonged delay to progression and more favorable effects on serum alkaline phosphatase.Conclusions: Although LHRH agonists are still the mainstay for treatment of advanced PCa, antagonists of LHRH offer an alternative as a pharmacological approach. (C) 2015 Elsevier Inc. All rights reserved.

Introduction: Increasing evidence suggests that prostate cancer cells undergo unique metabolic reprogramming during transformation. A master regulator of cellular homeostasis, 5'-AMP-activated protein kinase (AMPK), directs metabolic adaptation that supports the growth demands of rapidly dividing cancer cells. The utilization of AMPK as a therapeutic target may therefore provide an effective strategy in the treatment of prostate cancer.Areas covered: Our review describes the regulation of AMPK by androgens and upstream kinases including the calcium/calmodulin-dependent protein kinase kinase 2 (CaMKK2) in prostate cancer. Oncogenic, AMPK-regulated pathways that direct various metabolic processes are also addressed. Furthermore, we discuss the role of AMPK in growth arrest and autophagy as a potential survival pathway for cancer cells. In addition, by regulating non-metabolic pathways, AMPK may stimulate migration and mitosis. Finally, this review summarizes efforts to treat prostate cancer with pharmacological agents capable of modulating AMPK signaling.Expert opinion: Current research is primarily focused on developing drugs that activate AMPK as a treatment for prostate cancer. However, oncogenic aspects of AMPK signaling calls for caution about employing such therapies. We think that inhibitors of CaMKK2 or AMPK, or perhaps the modulation of downstream targets of AMPK, will gain importance in the clinical management of prostate cancer.

Papillary thyroid cancer (PTC) is the most prevalent of all endocrine cancers. In recent studies, the presence of receptors for pituitary-type growth hormone-releasing hormone (pGHRH-R) has been demonstrated in various human cancers, including human prostate, brain, and other cancer lines. Thyroid malignancies, however, have not yet been investigated in this regard. In this study, we found that pGHRH-R and its functional splice variant, SV1, are present in normal thyroid and PTC cells. We also treated seven normal and PTC tumor thyroid cells in vitro with a GHRH antagonist, MIA-602, to compare its anti-proliferation and anti-invasion potential against vehicle-treated cells. We found that treatment with GHRH antagonist increases the expression of SV1 and pGHRH-R in tumor cells compared to tumor cells exposed to vehicle only, a response which may alter the sensitivity of signaling kinases within the cells. GHRH antagonist treatment of tumor cells also reduced activity of the tumor invasion marker, matrix metalloproteinase (MMP)-2, compared to tumor cells exposed to vehicle only. The expression of pGHRH-R and SV1, as well as MMP-2 activity, in normal thyroid cells remained unaffected by GHRH antagonist treatment. Similarly, cell proliferation rates for tumor or normal thyroid cells were not affected by GHRH antagonist treatment. Our findings have important implications for the therapeutic use of GHRH antagonist in cases of aggressive PTC refractory to conventional treatment modalities, and in which protein expression and MMP-2 activity in normal thyroid tissue is left unaltered.

Malignant melanoma is the deadliest form of skin cancer; the treatment of advanced and recurrent forms remains a challenge. It has recently been reported that growth hormone-releasing hormone (GHRH) receptor is involved in the pathogenesis of melanoma. Therefore, we investigated the effects of our new GHRH antagonists on a human melanoma cancer cell line. Antiproliferative effects of GHRH antagonists, MIA-602, MIA-606 and MIA-690, on the human melanoma cell line, A-375, were studied in vitro using the MTS assay. The effect of MIA-690 (5 μg/day 28 d) was further evaluated in vivo in nude mice bearing xenografts of A-375. Subcellular localization of p27 was detected with Western blot and immunofluorescent staining. MIA-690 inhibited the proliferation of A-375 cells in a dose-dependent manner (33% at 10 μM, and 19.2% at 5 μM, P < 0 .05 vs. control), and suppressed the growth of xenografted tumors by 70.45% (P < 0.05). Flow cytometric analysis of cell cycle effects following the administration of MIA-690 revealed a decrease in the number of cells in G2/M phase (from 19.7% to 12.9%, P < 0.001). Additionally, Western blot and immunofluorescent studies showed that exposure of A-375 cells to MIA-690 triggered the nuclear accumulation of p27. MIA-690 inhibited tumor growth in vitro and in vivo, and increased the translocation of p27 into the nucleus thus inhibiting progression of the cell cycle. Our findings indicate that patients with malignant melanoma could benefit from treatment regimens, which combine existing chemotherapy agents and novel GHRH-antagonists.