Disclosures TRPV1 Mediates Anxious/Depressive No disclosures - - PDF document

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TRPV1 Mediates Anxious/Depressive Behavior in a Murine Model of Chronic Abdominal Pain

NASPGHAN Annual Meeting Oct 20, 2012

John Rosen, MD

Division of Pediatric Gastroenterology, Hepatology, and Nutrition Ann & Robert H. Lurie Children’s Hospital of Chicago Northwestern University Feinberg School of Medicine

Disclosures

• No disclosures
• No conflict of interest

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Introduction

• Gastrointestinal and genitourinary visceral

pain syndromes overlap

• Chronic GI and GU pain are associated

with anxiety and depression

• Mechanisms of chronic pain incompletely

understood

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TRPV1 is distributed widely

• Central and peripheral nervous system

– Brain – Spinal cord (especially lumbar) – DRG – C and Aδ sensory fibers (largely lamina I&II) – Cutaneous free nerve terminals

• Intestinal mucosa / myenteric plexus
• Urothelium

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Laboratory models suggest TRPV1 mediates allodynia

• Lack of TRPV1 inhibits cyclophosphamide cystitis-

induced increased mechanical sensitivity

(Wang et al., Pain, 2008)

• LPS sensitizes TRPV1 via activation of TLR4 in

trigeminal sensory neurons

(Diogenes et al., J Dental Res, 2011)

• LPS component of SΦ874 bacteria induces TLR4

dependent chronic pelvic allodynia

(Rudick et al., PLOS One, 2012)

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Hypothesis

TRPV1 mediates chronic pelvic allodynia in a murine model of infectious cystitis

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Methods

NU14 O18 SΦ874 wz* Source Pain UTI Lab Chronic Acute

100 200 300 10 20 30

S874 S874+pDGal

Pain (% increase)

Time (days)

Methods

• 10-12 wk old female C57BL/6 mice
• Transurethral infection with SΦ874

– Capsazepine (TRPV1 antagonist) or vehicle

• Pelvic von Frey test of referred visceral pain
• Novelty suppressed feeding (NSF) behavioral test
• Visceromotor response to bladder distension

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SΦ874 induced chronic allodynia is abrogated by early capsazepine

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SΦ874 induced chronic allodynia is abrogated by early capsazepine

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Post-infection Day 15

SΦ874 induced chronic anxiety is blunted by early capsazepine

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Post-infection Day 15

Preliminary VMR Data

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Summary

• SΦ874 cystitis is associated with chronic

increased anxious/depressive behavior

• Capsazepine administered at the time of

SΦ874 infection abrogates chronic pelvic allodynia and anxious/depressive behavior

• Capsazepine administered 2 weeks after

SΦ874 infection does not alter allodynia or behavior

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Conclusions

• TRPV1 may mediate chronic allodynia and

anxiety/depression induced by SΦ874 bacterial cystitis

• Induction, but not maintenance, of allodynia

and anxiety/depression may be mediated by TRPV1

• NSF behavior testing can be a useful adjunct

in models of chronic abdominal/pelvic pain

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Future Directions

• Other measures of visceral pain and behavior
• Increased specificity of TRPV1 antagonist
• Additional bacterial and viral models

– Human isolates – Acute and chronic pain

• Physiologic study of TRPV1 sensitization

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Acknowledgements

Northwestern University

Klumpp Laboratory David Klumpp, PhD Anthony Schaeffer, MD Charles Rudick, PhD Wenbin Yang, PhD Andrew Piotrowski, PhD Vladimir Pavlov, MD Lizzie Aguiniga Ryan Yaggie Yelena Radulovic, MD, PhD Behavioral Phenotyping Core Mouse Histology Laboratory

Medical College of Wisconsin

Adrian Miranda, MD

University of Pittsburgh Medical Center

Gerald Gebhart, PhD

Scholarship Committee

David Klumpp, PhD Miguel Saps, MD John Lavigne, PsyD

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Questions?

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SΦ874 induced chronic allodynia is abrogated by early capsazepine

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VMR data, add in background/methods if using this slide

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TRPV1

Nature Reviews Drug Discovery 6, 357-372 (May 2007)

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Lack of TRPV1 inhibits cystitis-induced increased mechanical sensitivity in mice

• Cyclophosphamide model of cystitis
• B6 vs. TRPV -/- mice

– Histologic evaluation of cystitis – Short-term urinary voiding frequency – Bladder capacity – Cystometry – Thermal and mechanical (VF) test of hindpaw

Wang, Bjorling, et al.. Pain. 2008; 139: 158-167.

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Wang, Bjorling, et al.. Pain. 2008; 139: 158-167.

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Wang, Bjorling, et al.. Pain. 2008; 139: 158-167.

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Neonatal cystitis-induced colonic hypersensitivity in adult rats: a model of viscero-visceral convergence

Miranda, Sengupta, et al.. Neurogastroenterology and Motility. 2011; 23: 683-e281.

• Zymosan model of cystitis
• Sprague-Dawley rats

– Bladder and colon histology – NR1 expression in spinal cord (western blot) – In vivo pelvic nerve afferent fiber recording – Visceromotor response to colorectal distension

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Miranda, Sengupta, et al.. Neurogastroenterology and Motility. 2011; 23: 683-e281.

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LPS Sensitizes TRPV1 via Activation of TLR4 in Trigeminal Sensory Neurons

Diogenes, Hargreaves, et al.. J Dental Research. 2011; 90: 759-764.

• Cultured trigeminal neurons
• Sprague-Dawley rats

– Immunofluorescence – LPS binding assay – Single cell calcium imaging – iCGRP release assay – Patch-clamp electrophysiology

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Murine Pelvic Pain Testing

• Von Frey filaments
• Response to stimulus

– Abdominal retraction – Licking or scratching – Jumping

• Specific to pelvic pain

– Hindpaw testing – Weight

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TRPV1 is increased in chronic noninflammatory pain

• Peritoneum of women with chronic pelvic pain
• Foci of endometriosis
• Irritable Bowel Syndrome (IBS)
• Inflammatory Bowel Disease + IBS

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Questions for me

• Why capsazepine instead of RTX etc?
• Distribution of SQ cap—BBB?
• Timing of SQ cap effect, metabolism?
• VF pelvic testing for referred pain?

Validated?

• Other behavioral tests?
• TRPV1 central action, psych?
• Need histology and urinary MPO to prove no

change in acute inflammation?

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