Changes in Inflammatory and Contractile Protein Expression in Patients With Painful Bladder Syndrome/IC.

Mediators of Neurogenic Inflammation in the Urinary Tract as Key Factors in the Painful Bladder Syndrome / Interstitial Cystitis and Bladder Dysfunction

Interstitial cystitis (IC)/chronic pelvic pain syndrome (CPPS) is a clinical syndrome of pelvic pain and/or urinary urgency/frequency in absence of a specific cause such as bacterial infection or damage to the bladder. The pathogenetic mechanisms of IC/CPPS are as yet undefined and it is largely this lack of knowledge, which precludes a systematic therapeutic approach. Experimental evidence, including results from the animal models of cystitis and the knock-out mice, indicate a participation of tachykinin receptors, especially the NK1R, in neurogenic inflammation, which is considered an important element of the IC complex. However, there is very scant information about the molecular mechanisms of IC in humans, or of the types of receptors, which participate in neurogenic inflammation. Based on our molecular biological know-how and the clinical expertise, we propose to investigate the role of the tachykinin and bradykinin receptors and their signalling partners in CPPS and bladder dysfunction in humans.

Study Overview

• Status: Completed
• Conditions: Overactive Bladder; Cystitis, Interstitial; Painful Bladder Syndrome; Chronic Pelvic Pain Syndrome

Detailed Description

Although IC has been recognised for more than a century, its pathophysiology remains a mystery, and as a consequence the treatment of IC is largely empirical. A multitude of mechanisms of the disease have been postulated ranging from neuroinflammatory to autoimmune or possibly infectious or toxic agents. Newer studies have hinted towards a genetic basis of the disease, but in most hypotheses an inflammatory component of some kind is involved and many findings support this theory. An often-cited hypothesis is the leaky epithelium. The healthy bladder is coated with a thin mucinous substance bladder surface mucin, which is composed of numerous sulfonated glycosaminoglycans and glycoproteins. In IC patients, in contrast to controls, as well as in some animal models of IC, qualitative changes to this surface layer have been observed (Lilly et al, 1990., Moskowitz et al, 1994). An initiating event (toxin) may lead to functional changes and increased permeability. This in turn leads to nerve sensitisation and possibly up-regulation. There is increasing evidence that the progression of IC is accompanied by a significant up-regulation of sensory nerves in the bladder (Letourneau, 1996). Nerve growth factor (NGF), a neurotropin that sensitises nociceptor fibres, was reported to be increased in bladders of IC patients (Lowe, 1997), and application of NGF in Wistar rats acutely induced bladder hyperactivity (Chuang, 2001). Sensory nerves in neurogenic inflammation secrete inflammatory mediators such as Substance P (SP), a nociceptive neurotransmitter in the central and peripheral nervous system. Increased amounts of the released Substance P have been found in the urine of IC patients (Hohenfellner, et al,1992, Pang et al., 1995, Pang et al., 1996), the concentration of Substance P reflecting the patients' degree of pain (Chen et al., 1999). Substance P and related tachykinins (TKs) mediate a variety of physiological processes in the genitourinary, pulmonary and gastrointestinal tract through stimulation of NK1 and NK2 receptors. Pre-clinical evidence obtained through use of selective tachykinin receptor antagonists indicates that endogenous tachykinins are involved in regulation of smooth muscle contraction, vasodilation, water metabolism and inflammation. Recently it was shown that mRNA encoding for SP receptor NK1 is in-creased in the bladder biopsies from patients with interstitial cystitis (Marchand et al., 1998). NK1R mRNA was found in detrusor muscle, urothelium and vascular structures, and the endothelial NK1R were markedly increased. SP binding to NK1R on endothelial cells results in vasodilation, plasma extravasation, cytokine release and activation and infiltration of immune cells, all characteristic of IC. Experiments with the NK1R knockout mice allowed for the first time to demonstrate the obligatory requirement of NK1R in cystitis and participation of these receptors in the chain of events linking mast cell degranulation and inflammation (Saban et al., 2000).

Changes in the sarcolemma, and altered expression of the tachykinin and bradykinin receptors might significantly influence the downstream signalling events leading to propagation of the symptoms of chronic pelvic pain/IC. By uncovering the link between clinical symptoms and the molecular regulation of the atypical inflammatory response, we may be able to offer novel and specific options for treatment. In the current proposal, we would like to further our studies of urinary bladder pathology, in particular the symptomatic complex of PBS/interstitial cystitis, and investigate the effect of changes in human urothelium and detrusor muscle cells on the NK1R-mediated signalling.

Biopsies from controls and PBS patients will be obtained in either general or spinal anesthesia transurethrally from the bladder dome and trigone with a biopsy tong. RNA will be extracted, and the expression levels of selected genes analyzed using Taqman real-time PCR and gene expression arrays (Applied Biosystems). Calcium imaging will be used to monitor the receptor activation and protein levels analysed by SDS-PAGE and Western Blotting. Receptor tissue distribution will be analysed by immuno-cytochemistry.

• Study Type: Observational
• Enrollment (Anticipated): 45

Contacts and Locations

Study Locations

• Switzerland
  • Bern, Switzerland, 3010
    • Dept. of Urology, Bern University Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (ADULT, OLDER_ADULT)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

• Sampling Method: Non-Probability Sample

Study Population

university hospital clinic

Description

Inclusion Criteria:

• Clinical asymptomatic bladder function
• Clinical urgency frequency syndrome/ overactive bladder
• Clinical painful bladder syndrome/interstitial cystitis

Exclusion Criteria:

• Not able to understand patient information
• Other cause of symptoms
• Confusable diseases

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort
1
2
3

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Time Frame
Receptor expression in disease states versus control	at time of biospy

Collaborators and Investigators

• Sponsor: University Hospital Inselspital, Berne

Publications and helpful links

General Publications

• Lilly JD, Parsons CL. Bladder surface glycosaminoglycans is a human epithelial permeability barrier. Surg Gynecol Obstet. 1990 Dec;171(6):493-6.
• Letourneau R, Pang X, Sant GR, Theoharides TC. Intragranular activation of bladder mast cells and their association with nerve processes in interstitial cystitis. Br J Urol. 1996 Jan;77(1):41-54. doi: 10.1046/j.1464-410x.1996.08178.x.
• Lowe EM, Anand P, Terenghi G, Williams-Chestnut RE, Sinicropi DV, Osborne JL. Increased nerve growth factor levels in the urinary bladder of women with idiopathic sensory urgency and interstitial cystitis. Br J Urol. 1997 Apr;79(4):572-7. doi: 10.1046/j.1464-410x.1997.00097.x.
• Chuang YC, Fraser MO, Yu Y, Chancellor MB, de Groat WC, Yoshimura N. The role of bladder afferent pathways in bladder hyperactivity induced by the intravesical administration of nerve growth factor. J Urol. 2001 Mar;165(3):975-9.
• Hohenfellner M, Nunes L, Schmidt RA, Lampel A, Thuroff JW, Tanagho EA. Interstitial cystitis: increased sympathetic innervation and related neuropeptide synthesis. J Urol. 1992 Mar;147(3):587-91. doi: 10.1016/s0022-5347(17)37314-7.
• Pang X, Marchand J, Sant GR, Kream RM, Theoharides TC. Increased number of substance P positive nerve fibres in interstitial cystitis. Br J Urol. 1995 Jun;75(6):744-50. doi: 10.1111/j.1464-410x.1995.tb07384.x.
• Pang X, Boucher W, Triadafilopoulos G, Sant GR, Theoharides TC. Mast cell and substance P-positive nerve involvement in a patient with both irritable bowel syndrome and interstitial cystitis. Urology. 1996 Mar;47(3):436-8. doi: 10.1016/S0090-4295(99)80469-5.
• Marchand JE, Sant GR, Kream RM. Increased expression of substance P receptor-encoding mRNA in bladder biopsies from patients with interstitial cystitis. Br J Urol. 1998 Feb;81(2):224-8. doi: 10.1046/j.1464-410x.1998.00507.x.
• Moskowitz MO, Byrne DS, Callahan HJ, Parsons CL, Valderrama E, Moldwin RM. Decreased expression of a glycoprotein component of bladder surface mucin (GP1) in interstitial cystitis. J Urol. 1994 Feb;151(2):343-5. doi: 10.1016/s0022-5347(17)34944-3.
• Saban R, Saban MR, Nguyen NB, Lu B, Gerard C, Gerard NP, Hammond TG. Neurokinin-1 (NK-1) receptor is required in antigen-induced cystitis. Am J Pathol. 2000 Mar;156(3):775-80. doi: 10.1016/S0002-9440(10)64944-9.

Study record dates

Study Major Dates

• Study Start: October 1, 2006
• Primary Completion (ACTUAL): April 1, 2010
• Study Completion (ACTUAL): April 1, 2010

Study Registration Dates

• First Submitted: October 17, 2008
• First Submitted That Met QC Criteria: October 17, 2008
• First Posted (ESTIMATE): October 20, 2008

Study Record Updates

• Last Update Posted (ESTIMATE): April 27, 2010
• Last Update Submitted That Met QC Criteria: April 25, 2010
• Last Verified: April 1, 2010

More Information

Terms related to this study

• Keywords: Inflammatory proteins; Tachykinin receptors; Protein expression; Contractile proteins
• Additional Relevant MeSH Terms: Pathologic Processes; Urologic Diseases; Urinary Bladder Diseases; Lower Urinary Tract Symptoms; Urological Manifestations; Neurologic Manifestations; Disease; Urinary Bladder, Overactive; Syndrome; Cystitis; Pain; Cystitis, Interstitial; Pelvic Pain
• Other Study ID Numbers: KEK_146_05