Growth hormone-releasing hormone antagonists reduce prostatic enlargement and inflammation in carrageenan-induced chronic prostatitis
Corresponding Author
Petra Popovics PhD
Division of Endocrinology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Correspondence
Petra Popovics, PhD, Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, FL, Research Service (151), 1201 NW 16th St, Miami, FL 33125.
Email: [email protected]
Search for more papers by this authorRenzhi Cai PhD
Division of Endocrinology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Search for more papers by this authorWei Sha BSc
Division of Endocrinology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Search for more papers by this authorFerenc G. Rick MD, PhD
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Department of Urology, Herbert Wertheim College of Medicine, Florida International, University, Miami, Florida
Search for more papers by this authorAndrew V. Schally PhD, MDhc (Multi), DSchc
Division of Endocrinology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Sylvester Comprehensive Cancer Center, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Division of Hematology/Oncology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Department of Pathology, Miller School of Medicine, University of Miami, Miami, Florida
Search for more papers by this authorCorresponding Author
Petra Popovics PhD
Division of Endocrinology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Correspondence
Petra Popovics, PhD, Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, FL, Research Service (151), 1201 NW 16th St, Miami, FL 33125.
Email: [email protected]
Search for more papers by this authorRenzhi Cai PhD
Division of Endocrinology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Search for more papers by this authorWei Sha BSc
Division of Endocrinology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Search for more papers by this authorFerenc G. Rick MD, PhD
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Department of Urology, Herbert Wertheim College of Medicine, Florida International, University, Miami, Florida
Search for more papers by this authorAndrew V. Schally PhD, MDhc (Multi), DSchc
Division of Endocrinology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Endocrine, Polypeptide and Cancer Institute, Veterans Affairs Medical Center, Miami, Florida
Sylvester Comprehensive Cancer Center, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Division of Hematology/Oncology, Department of Medicine, Miller School of Medicine, University of Miami, Miami, Florida
Department of Pathology, Miller School of Medicine, University of Miami, Miami, Florida
Search for more papers by this authorAbstract
Background
Inflammation 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.
Methods
To 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.
Results
Inflammation 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.
Conclusions
The 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.
CONFLICTS OF INTEREST
No competing financial interests exist. The authors have nothing to disclose.
Supporting Information
Additional supporting information may be found online in the Supporting Information section at the end of the article.
| Filename | Description |
|---|---|
| pros23655-sup-0001-SuppFig-S1.docx3.5 MB |
Fig. S1. Immunohistological localization of GHRH, TGF-β1, IGF-1, matrix metalloproteinase 9 (MMP9) and osteopontin in the ventral lobes of normal and inflamed prostates. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
REFERENCES
- 1 Roehrborn CG. Benign prostatic hyperplasia: an overview. Rev Urol. 2005; 7: S3–S14.
- 2 McNeal JE. Origin and evolution of benign prostatic enlargement. Invest Urol. 1978; 15: 340–345.
- 3 Hutchison A, Farmer R, Chapple C, et al. Characteristics of patients presenting with LUTS/BPH in six European countries. Eur Urol. 2006; 50: 555–561. discussion 562.
- 4 Vuichoud C, Loughlin KR. Benign prostatic hyperplasia: epidemiology, economics and evaluation. Can J Urol. 2015; 22: 1–6.
- 5 McVary KT, Roehrborn CG, Avins AL, et al. Update on AUA guideline on the management of benign prostatic hyperplasia. J Urol. 2011; 185: 1793–1803.
- 6 Slater S, Dumas C, Bubley G. Dutasteride for the treatment of prostate-related conditions. Expert Opin Drug Saf. 2012; 11: 325–330.
- 7 Steers WD. 5alpha-reductase activity in the prostate. Urology. 2001; 58: 17–24. discussion 24.
- 8 Singh MK, Avram M. Persistent sexual dysfunction and depression in finasteride users for male pattern hair loss: a serious concern or red herring? J Clin Aesthet Dermatol. 2014; 7: 51–55.
- 9 Kaplan SA. Side Effects of alpha-Blocker use: retrograde ejaculation. Rev Urol. 2009; 11: S14–S18.
- 10 Schoenfeld MJ, Shortridge EF, Gelwicks SC, Cui Z, Wong DG. Treatment patterns in alpha-blocker therapy for benign prostatic hyperplasia. Am J Mens Health. 2014; 8: 267–272.
- 11 Peixoto CA, Gomes FO. The role of phosphodiesterase-5 inhibitors in prostatic inflammation: a review. J Inflamm (Lond). 2015; 12: 54.
- 12 Cunha GR, Ricke W, Thomson A, et al. Hormonal, cellular, and molecular regulation of normal and neoplastic prostatic development. J Steroid Biochem Mol Biol. 2004; 92: 221–236.
- 13 Untergasser G, Madersbacher S, Berger P. Benign prostatic hyperplasia: age-related tissue-remodeling. Exp Gerontol. 2005; 40: 121–128.
- 14
Shibata Y,
Ito K,
Suzuki K, et al. Changes in the endocrine environment of the human prostate transition zone with aging: simultaneous quantitative analysis of prostatic sex steroids and comparison with human prostatic histological composition.
Prostate.
2000;
42: 45–55.
10.1002/(SICI)1097-0045(20000101)42:1<45::AID-PROS6>3.0.CO;2-W CAS PubMed Web of Science® Google Scholar
- 15 Hammarsten J, Hogstedt B, Holthuis N, Mellstrom D. Components of the metabolic syndrome-risk factors for the development of benign prostatic hyperplasia. Prostate Cancer Prostatic Dis. 1998; 1: 157–162.
- 16 Gandaglia G, Briganti A, Gontero P, et al. The role of chronic prostatic inflammation in the pathogenesis and progression of benign prostatic hyperplasia (BPH). BJU Int. 2013; 112: 432–441.
- 17 Kohnen PW, Drach GW. Patterns of inflammation in prostatic hyperplasia: a histologic and bacteriologic study. J Urol. 1979; 121: 755–760.
- 18 Wang X, Lin WJ, Izumi K, et al. Increased infiltrated macrophages in benign prostatic hyperplasia (BPH): role of stromal androgen receptor in macrophage-induced prostate stromal cell proliferation. J Biol Chem. 2012; 287: 18376–18385.
- 19 Xu D, Wang X, Jiang C, Ruan Y, Xia S, Wang X. The androgen receptor plays different roles in macrophage-induced proliferation in prostate stromal cells between transitional and peripheral zones of benign prostatic hypertrophy. EXCLI J. 2017; 16: 939–948.
- 20 Lu T, Lin WJ, Izumi K, et al. Targeting androgen receptor to suppress macrophage-induced EMT and benign prostatic hyperplasia (BPH) development. Mol Endocrinol. 2012; 26: 1707–1715.
- 21 Popovics P, Schally AV, Salgueiro L, Kovacs K, Rick FG. Antagonists of growth hormone-releasing hormone inhibit proliferation induced by inflammation in prostatic epithelial cells. Proc Natl Acad Sci U S A. 2017; 114: 1359–1364.
- 22 McDowell KL, Begley LA, Mor-Vaknin N, Markovitz DM, Macoska JA. Leukocytic promotion of prostate cellular proliferation. Prostate. 2010; 70: 377–389.
- 23 Begley LA, Kasina S, MacDonald J, Macoska JA. The inflammatory microenvironment of the aging prostate facilitates cellular proliferation and hypertrophy. Cytokine. 2008; 43: 194–199.
- 24 Shao R, Shi J, Liu H, et al. Epithelial-to-mesenchymal transition and estrogen receptor alpha mediated epithelial dedifferentiation mark the development of benign prostatic hyperplasia. Prostate. 2014; 74: 970–982.
- 25 Muller EE, Locatelli V, Cocchi D. Neuroendocrine control of growth hormone secretion. Physiol Rev. 1999; 79: 511–607.
- 26 Kiaris H, Chatzistamou I, Papavassiliou AG, Schally AV. Growth hormone-releasing hormone: not only a neurohormone. Trends Endocrinol Metab. 2011; 22: 311–317.
- 27 Rick FG, Schally AV, Szalontay L, et al. Antagonists of growth hormone-releasing hormone inhibit growth of androgen-independent prostate cancer through inactivation of ERK and Akt kinases. Proc Natl Acad Sci U S A. 2012; 109: 1655–1660.
- 28
Schally AV,
Comaru-Schally AM,
Plonowski A,
Nagy A,
Halmos G,
Rekasi Z. Peptide analogs in the therapy of prostate cancer.
The Prostate.
2000;
45: 158–166.
10.1002/1097-0045(20001001)45:2<158::AID-PROS10>3.0.CO;2-K CAS PubMed Web of Science® Google Scholar
- 29 Schally AV, Varga JL, Engel JB. Antagonists of growth-hormone-releasing hormone: an emerging new therapy for cancer. Nat Clin Pract Endocrinol Metab. 2008; 4: 33–43.
- 30 Fahrenholtz CD, Rick FG, Garcia MI, et al. Preclinical efficacy of growth hormone-releasing hormone antagonists for androgen-dependent and castration-resistant human prostate cancer. Proc Natl Acad Sci U S A. 2014; 111: 1084–1089.
- 31 Zarandi M, Cai R, Kovacs M, et al. Synthesis and structure-activity studies on novel analogs of human growth hormone releasing hormone (GHRH) with enhanced inhibitory activities on tumor growth. Peptides. 2017; 89: 60–70.
- 32 Rick FG, Schally AV, Block NL, et al. Antagonists of growth hormone-releasing hormone (GHRH) reduce prostate size in experimental benign prostatic hyperplasia. Proc Natl Acad Sci U S A. 2011; 108: 3755–3760.
- 33 Rick FG, Szalontay L, Schally AV, et al. Combining growth hormone-releasing hormone antagonist with luteinizing hormone-releasing hormone antagonist greatly augments benign prostatic hyperplasia shrinkage. J Urol. 2012; 187: 1498–1504.
- 34 Rick FG, Schally AV, Block NL, Abi-Chaker A, Krishan A, Szalontay L. Mechanisms of synergism between antagonists of growth hormone-releasing hormone and antagonists of luteinizing hormone-releasing hormone in shrinking experimental benign prostatic hyperplasia. Prostate. 2013; 73: 873–883.
- 35 Zarandi M, Horvath JE, Halmos G, et al. Synthesis and biological activities of highly potent antagonists of growth hormone-releasing hormone. Proc Natl Acad Sci U S A. 1994; 91: 12298–12302.
- 36 Zeng F, Chen H, Yang J, et al. Development and validation of an animal model of prostate inflammation-induced chronic pelvic pain: evaluating from inflammation of the prostate to pain behavioral modifications. PLoS ONE. 2014; 9: e96824.
- 37 Jerde TJ, Bushman W. IL-1 induces IGF-dependent epithelial proliferation in prostate development and reactive hyperplasia. Sci Signal. 2009; 2: ra49.
- 38 Penna G, Fibbi B, Amuchastegui S, et al. Human benign prostatic hyperplasia stromal cells as inducers and targets of chronic immuno-mediated inflammation. J Immunol. 2009; 182: 4056–4064.
- 39 Wang HH, Wang L, Jerde TJ, et al. Characterization of autoimmune inflammation induced prostate stem cell expansion. Prostate. 2015; 75: 1620–1631.
- 40 Rodriguez-Nieves JA, Patalano SC, Almanza D, Gharaee-Kermani M, Macoska JA. CXCL12/CXCR4 axis activation mediates prostate myofibroblast phenoconversion through non-canonical EGFR/MEK/ERK signaling. PLoS ONE. 2016; 11: e0159490.
- 41 Vykhovanets EV, Resnick MI, MacLennan GT, Gupta S. Experimental rodent models of prostatitis: limitations and potential. Prostate Cancer Prostatic Dis 2007; 10: 15–29.
- 42 Necas J, Bartosikova L. Carrageenan: a review. Veterinarni Medicina. 2013; 58: 187–205.
- 43 Borthakur A, Bhattacharyya S, Dudeja PK, Tobacman JK. Carrageenan induces interleukin-8 production through distinct Bcl10 pathway in normal human colonic epithelial cells. Am J Physiol Gastrointest Liver Physiol. 2007; 292: G829–G838.
- 44 Bhattacharyya S, Gill R, Chen ML, et al. Toll-like receptor 4 mediates induction of the Bcl10-NFkappaB-interleukin-8 inflammatory pathway by carrageenan in human intestinal epithelial cells. J Biol Chem. 2008; 283: 10550–10558.
- 45 Wu W, Zhen Z, Niu T, et al. Kappa-carrageenan enhances lipopolysaccharide-induced interleukin-8 secretion by stimulating the Bcl10-NF-kappaB pathway in HT-29 cells and aggravates C. freundii-induced inflammation in mice. Mediators Inflamm. 2017; 2017: 8634865.
- 46 Chen CS, Chang PJ, Lin WY, Huang YC, Ho DR. Evidences of the inflammasome pathway in chronic prostatitis and chronic pelvic pain syndrome in an animal model. Prostate. 2013; 73: 391–397.
- 47 Gatti G, Quintar AA, Andreani V, et al. Expression of Toll-like receptor 4 in the prostate gland and its association with the severity of prostate cancer. Prostate 2009; 69: 1387–1397.
- 48 Hamakawa T, Sasaki S, Shibata Y, et al. Interleukin-18 may lead to benign prostatic hyperplasia via thrombospondin-1 production in prostatic smooth muscle cells. Prostate. 2014; 74: 590–601.
- 49 Gan J, Ke X, Jiang J, et al. Growth hormone-releasing hormone receptor antagonists inhibit human gastric cancer through downregulation of PAK1-STAT3/NF-kappaB signaling. Proc Natl Acad Sci U S A. 2016; 113: 14745–14750.
- 50 Borges FT, Melo SA, Ozdemir BC, et al. TGF-beta1-containing exosomes from injured epithelial cells activate fibroblasts to initiate tissue regenerative responses and fibrosis. J Am Soc Nephrol. 2013; 24: 385–392.
- 51 Macoska J. Prostatic fibrosis is associated with lower urinary tract symptoms. J Urol. 2012; 188: 1375–1381.
- 52 Munoz-Moreno L, Bajo AM, Prieto JC, Carmena MJ. Growth hormone-releasing hormone (GHRH) promotes metastatic phenotypes through EGFR/HER2 transactivation in prostate cancer cells. Mol Cell Endocrinol. 2017; 446: 59–69.
- 53 Munoz-Moreno L, Arenas MI, Carmena MJ, Schally AV, Prieto JC, Bajo AM. Growth hormone-releasing hormone antagonists abolish the transactivation of human epidermal growth factor receptors in advanced prostate cancer models. Invest New Drugs. 2014; 32: 871–882.
- 54 Vacas E, Munoz-Moreno L, Valenzuela PL, et al. Growth hormone-releasing hormone induced transactivation of epidermal growth factor receptor in human triple-negative breast cancer cells. Peptides. 2016; 86: 153–161.
- 55 Hayward SW, Dahiya R, Cunha GR, Bartek J, Deshpande N, Narayan P. Establishment and characterization of an immortalized but non-transformed human prostate epithelial cell line: BPH-1. In Vitro Cell Dev Biol Anim. 1995; 31: 14–24.
- 56 Corsi MM, Fulgenzi A, Gaja G, et al. Effect of acute and chronic inflammation on plasma growth hormone levels in rats. Drugs Exp Clin Res. 1997; 23: 117–122.
- 57 Cardillo MR, Monti S, Di Silverio F, Gentile V, Sciarra F, Toscano V. Insulin-like growth factor (IGF)-I, IGF-II and IGF type I receptor (IGFR-I) expression in prostatic cancer. Anticancer Res. 2003; 23: 3825–3835.
- 58 Zhao D, Bakirtzi K, Zhan Y, Zeng H, Koon HW, Pothoulakis C. Insulin-like growth factor-1 receptor transactivation modulates the inflammatory and proliferative responses of neurotensin in human colonic epithelial cells. J Biol Chem. 2011; 286: 6092–6099.
- 59 Yabluchanskiy A, Ma Y, Iyer RP, Hall ME, Lindsey ML. Matrix metalloproteinase-9: many shades of function in cardiovascular disease. Physiology (Bethesda). 2013; 28: 391–403.
- 60 Cantor H, Shinohara ML. Regulation of T-helper-cell lineage development by osteopontin: the inside story. Nat Rev Immunol. 2009; 9: 137–141.
- 61 Kourepini E, Aggelakopoulou M, Alissafi T, Paschalidis N, Simoes DC, Panoutsakopoulou V, Osteopontin expression by CD103-dendritic cells drives intestinal inflammation. Proc Natl Acad Sci U S A. 2014; 111: E856–865.
