mMCP7, a Mouse Ortholog of δ Tryptase, Mediates Pelvic Tactile Allodynia in a Model of Chronic Pelvic Pain

Chronic prostatitis/Chronic pelvic pain syndrome (CP/CPPS) is a condition that affects a large number of men and has unknown etiology. We have previously demonstrated the presence of elevated levels of mast cell tryptase in expressed prostatic secretions (EPS) of CP/CPPS patients. In a murine model of CP/CPPS, we showed tryptase and its cognate receptor PAR2 as critical to the development of pelvic pain and lower urinary tract symptoms. Here, we extend these observations to demonstrate that an isoform of tryptase called delta (δ)-tryptase, is elevated in the EPS of patients with CP/CPPS and is correlated with pelvic pain symptoms. Using an Escherichia coli (CP1) -induced murine model of CP/CPPS, we demonstrated a differential response in C57BL/6J and NOD/ShiLtJ mice, with C57BL6/J mice being resistant to an increase in pelvic tactile allodynia, despite having equivalent levels of activated mast cells in the prostate. Activated tryptase+ve mast cells were observed to be in closer apposition to PGP9.5+ve nerve fibers in the prostate stroma of NOD/ShiLtJ in comparison to C57BL/6J mice. The mouse ortholog of δ-tryptase, mouse mast cell protease 7 (mMCP7) has been reported to be unexpressed in C57BL/6J mice. We confirmed the absence of mMCP7 in the prostates of C57BL/6J and its presence in NOD/ShiLtJ mice. To evaluate a role for mMCP7 in the differential allodynia responses, we performed direct intra-urethral instillations of mMCP7 and the beta (β)-tryptase isoform ortholog, mMCP6 in the CP1-infection model. mMCP7, but not mMCP6 was able to induce an acute pelvic allodynia response in C57BL/6J mice. In-vitro studies with mMCP7 on cultured mast cells as well as dissociated primary neurons demonstrated the ability to induce differential activation of pain and inflammation associated molecules compared to mMCP6. We conclude that mMCP7, and possibility its human ortholog δ-tryptase, may play an important role in mediating the development of pelvic tactile allodynia in the mouse model of pelvic pain and in patients with CP/CPPS.

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A novel murine model of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) induced by immunization with a spermine binding protein (p25) peptide

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00147.2012 ◽

2013 ◽

Vol 304 (6) ◽

pp. R415-R422 ◽

Cited By ~ 12

Author(s):

Cengiz Z. Altuntas ◽

Firouz Daneshgari ◽

Elias Veizi ◽

Kenan Izgi ◽

Fuat Bicer ◽

...

Keyword(s):

Pelvic Pain ◽

Murine Model ◽

Chronic Prostatitis ◽

Chronic Pelvic Pain ◽

Chronic Pelvic Pain Syndrome ◽

Urine Volume ◽

Pain Syndrome ◽

Pelvic Pain Syndrome ◽

Human Phenotype ◽

Micturition Frequency

The pathophysiology of chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is poorly understood. Inflammatory and autoimmune mechanisms may play a role. We developed a murine model of experimental autoimmune prostatitis (EAP) that mimics the human phenotype of CP/CPPS. Eight-week-old mice were immunized subcutaneously with prostate-specific peptides in an emulsion of complete Freund's adjuvant. Mice were euthanized 10 days after immunization, and lymph node cells were isolated and assessed for recall proliferation to each peptide. P25 99–118 was the most immunogenic peptide. T-cell and B-cell immunity and serum levels of C-reactive protein and nitrate/nitrite levels were evaluated over a 9-wk period. Morphometric studies of prostate, 24-h micturition frequencies, and urine volume per void were evaluated. Tactile referred hyperalgesia was measured using von Frey filaments to the pelvic region. The unpaired Student's t-test was used to analyze differences between EAP and control groups. Prostates from p25 99–118-immunized mice demonstrated elevated gene expression levels of TNF-α, IL-17A, IFN-γ, and IL-1β, not observed in control mice. Compared with controls, p25 99–118-immunized mice had significantly higher micturition frequency and decreased urine output per void, and they demonstrated elevated pelvic pain response. p25 99–118 immunization of male SWXJ mice induced prostate-specific autoimmunity characterized by prostate-confined inflammation, increased micturition frequency, and pelvic pain. This autoimmune prostatitis model provides a useful tool for exploring the pathophysiology and new treatments.

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Microbiota of Chronic Prostatitis/Chronic Pelvic Pain Syndrome are Distinct from Interstitial Cystitis/Bladder Pain Syndrome

10.1101/2021.03.04.21252926 ◽

2021 ◽

Author(s):

Bryan White ◽

Michael Welge ◽

Loretta Auvil ◽

Matthew Berry ◽

Colleen Bushell ◽

...

Keyword(s):

Interstitial Cystitis ◽

Pelvic Pain ◽

Chronic Prostatitis ◽

Chronic Pelvic Pain ◽

Chronic Pelvic Pain Syndrome ◽

Pain Syndrome ◽

Bladder Pain Syndrome ◽

Unknown Etiology ◽

Pelvic Pain Syndrome ◽

Bladder Pain

Urologic chronic pelvic pain syndrome patients include men chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) and patients, mainly women, with interstitial cystitis/bladder pain syndrome (IC/BPS or IC). CP/CPPS is marked by severe chronic pelvic pain of unknown etiology that is differentially associated with prostatic inflammation. Microbes are known to modulate sensory responses, and microbiota are increasingly understood to drive normal biological processes and pathogenesis, including inflammation. Recent studies have linked fecal dysbiosis with chronic pelvic pain in IC/BPS, suggesting a role for microbiota in modulating UCPPS pain. Similarly, dysbiosis has been reported in CP/CPPS patients, but the relationship between with the dysbiosis of IC/BPS patients is unclear. Here, we characterized the fecal microbiota of men with CP/CPPS and women and men with IC/BPS. Similar to recent reports, we identified fecal dysbiosis in men with CP/CPPS relative to healthy controls among specific phyla and overall differences in diversity and richness. Interestingly, we also observed differences between CP/CPPS microbiota and IC/BPS microbiota that were not likely due to sex differences. These findings suggest that CP/CPPS is marked by changes in the gut microbiome, but these changes differ from IC/BPS. Taken together, UCPPS appears associated with distinct dybioses among CP/CPPS and IC/BPS, raising the possibility of distinct contributions to underlying pelvic pain mechanisms and/or etiologies.

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UropathogenicEscherichia coliInduces Chronic Pelvic Pain

Infection and Immunity ◽

10.1128/iai.00910-10 ◽

2010 ◽

Vol 79 (2) ◽

pp. 628-635 ◽

Cited By ~ 60

Author(s):

Charles N. Rudick ◽

Ruth E. Berry ◽

James R. Johnson ◽

Brian Johnston ◽

David J. Klumpp ◽

...

Keyword(s):

Pelvic Pain ◽

Chronic Pelvic Pain ◽

Prostate Gland ◽

Chronic Pelvic Pain Syndrome ◽

Nod Mice ◽

Pain Syndrome ◽

Unknown Etiology ◽

Pelvic Pain Syndrome ◽

Microbial Infection ◽

Microbial Infections

ABSTRACTChronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a debilitating syndrome of unknown etiology often postulated, but not proven, to be associated with microbial infection of the prostate gland. We hypothesized that infection of the prostate by clinically relevant uropathogenicEscherichia coli(UPEC) can initiate and establish chronic pain. We utilized anE. colistrain newly isolated from a patient with CP/CPPS (strain CP1) and examined its molecular pathogenesis in cell culture and in a murine model of bacterial prostatitis. We found that CP1 is an atypical isolate distinct from most UPEC in its phylotype and virulence factor profile. CP1 adhered to, invaded, and proliferated within prostate epithelia and colonized the prostate and bladder of NOD and C57BL/6J mice. Using behavioral measures of pelvic pain, we showed that CP1 induced and sustained chronic pelvic pain in NOD mice, an attribute not exhibited by a clinical cystitis strain. Furthermore, pain was observed to persist even after bacterial clearance from genitourinary tissues. CP1 induced pelvic pain behavior exclusively in NOD mice and not in C57BL/6J mice, despite comparable levels of colonization and inflammation. Microbial infections can thus serve as initiating agents for chronic pelvic pain through mechanisms that are dependent on both the virulence of the bacterial strain and the genetic background of the host.

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Pathological neurogenesis: a key step in the pathogenesis of pelvic pain caused by adenomyosis

ZHurnal «Patologicheskaia fiziologiia i eksperimental`naia terapiia» ◽

10.25557/0031-2991.2018.01.59-64 ◽

2018 ◽

pp. 59-64

Author(s):

М.Р. Оразов ◽

В.Е. Радзинский ◽

М.Б. Хамошина ◽

Е.Н. Носенко ◽

Т.В. Галина ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Pelvic Pain ◽

Hormonal Therapy ◽

Chronic Pelvic Pain ◽

Chronic Pelvic Pain Syndrome ◽

Pain Syndrome ◽

Nerve Fibers ◽

Pelvic Pain Syndrome ◽

Neurofilament Proteins ◽

Ectopic Endometrium

Статья посвящена одной из актуальных проблем клинической гинекологии - тазовой боли при аденомиозе. Цель исследования: изучение морфологических особенностей нервного аппарата миометрия у женщин с синдромом хронической тазовой боли, обусловленной аденомиозом. Методика. Изучено 60 биоптатов, полученных после гистерэктомий у женщин с диффузным аденомиозом ІІ-ІІІ степени, сопровождающимся болевым синдромом выраженной степени тяжести, не получавших гормональную терапию. Группа морфологического сравнения составила 10 биоптатов, полученных у женщин с аденомиозом, без болевого синдрома, прооперированных по поводу аномальных маточных кровотечений, также не получавших гормональную терапию. У всех пациентов получено информированное согласие на использование биологического материала в исследовании. После гистерэктомии участки стенок маток, включавшие эндометрий и миометрий подвергали стандартным гистологическим процедурам, готовили парафиновые срезы (5 мкм). Общую морфологическую оценку проводили при окраске срезов гематоксилином и эозином. Визуализацию нервных волокон осуществляли после иммуногистохимического (ИГХ) окрашивания с использованием моноклональных антител (МАТ) к белкам нейрофиламентов (DAKO, Дания) по инструкциям фирмы. Результаты. Изучение иннервационного аппарата матки при аденомиозе с помощью моноклональных антител к белкам нейрофиламентов выявил разнообразие волокон по толщине, плотности и интенсивности окраски. Плотность нервных окончаний, сопряженных непосредственно с очагами эндометриоза, была невелика - в зоне желез 4,1 ± 0,3 на мм среза, в строме - 9,2 ± 0,6. Тонкие нервные волокна визуализировались преимущественно в строме вокруг кровеносных сосудов, сопровождающих зоны врастания эктопического эндометрия. Густое сплетение тонких нервных волокон также было обнаружено в субсерозном слое. Причем, как в субсерозном, так и в подслизистом слое миометрия превалировали разветвленные тонкие нервные волокна, количество которых статистически значимо (p<0,01) превышало аналогичный показатель в группе сравнения - 17,2 ± 1,4 против 11,8 ± 0,9 на мм. Сравнение иннервационного аппарата матки у женщин контрольной группы и у пациенток с аденомиозом с болевым синдромом позволяет утверждать, что именно расширение иннервационного поля в миометрии является наиболее вероятной причиной формирования тазовой боли у пациенток с аденомиозом. Выводы. Результаты проделанной работы продемонстрировали, что основным местом локализации нервов в матке и потенциальной причиной формирования гипералгезии при аденомиозе является миометрий с формированием аномально избыточного иннервационного аппарата вокруг очагов эктопического эндометрия, в периваскулярных регионах и в строме между пучками гладких миоцитов. Objective. To study morphological features of the myometrium neural apparatus in women with chronic pelvic pain syndrome associated with adenomyosis. Methods. 60 biopsy samples were studied, which were obtained from hysterectomies for grade I-III diffuse adenomyosis associated with severe pelvic pain syndrome. These women did not receive any hormonal therapy. The control group included 10 biopsies obtained from women with adenomyosis who had no pelvic pain syndrome and underwent a surgery for abnormal uterine bleeding. These women did not receive any hormonal therapy either. After hysterectomy, uterine wall samples including endometrium and myometrium were fixed in 10% neutral, buffered formalin (pH 7.4) for 24 hours. After dehydration, the material was embedded in paraffin highly purified with polymeric additives (Richard-Allan Scientific, USA) at a temperature <60°C. Overall morphological evaluation of sections was performed with H E staining. Imaging was performed after nerve fibers immunohistochemical (IHC) staining with monoclonal antibodies (MAbs) to neurofilament proteins (DAKO, Denmark) according to the manufacturer’s instructions. Results. Studying of the uterine innervation apparatus using monoclonal antibodies to neurofilament proteins in adenomyosis identified a variety of fiber thickness, density and color intensity. Density of nerve endings directly associated with endometriosis foci was low; 4.1 ± 0.3 per mm in the glandular zone and 9.2 ± 0.6 per mm in the stroma. Thin nerve fibers were visualized mainly in the stroma around blood vessels associated with the ectopic endometrium ingrowth zone. A dense plexus of thin nerve fibers was also found in the subserosal layer. Furthermore, both in myometrial subserosal and submucosal layers, ramified, thin nerve fibers predominated. The number of such fibers was significantly greater than in the comparison group (17.2 ± 1.4 vs. 11.8 ± 0.9 per mm, p<0.01). Comparison of the uterine innervation apparatus in women with and without chronic pelvic pain syndrome suggested that particularly the expansion of myometrium innervation field was the most likely cause for the development of pelvic pain in women with adenomyosis. Conclusions. The main location of uterine nerves and a potential cause for hyperalgesia in adenomyosis is the myometrium developing abnormally excessive innervation around foci of ectopic endometrium, perivascular areas, and the stroma between fibers of smooth myocytes.

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