Background & Aims

Persistent abdominal pain (PAP) is cited as one of the most impactful symptoms across many disorders of gut-brain interactions (DGBIs) including irritable bowel syndrome (IBS). Visceral hypersensitivity (VH) is a primary driver of chronic AP in DGBIs, regardless of any co-occurring alterations in bowel habits. PAP is the most troublesome symptom reported by IBS patients, but most treatments primarily target bowel habits rather than addressing VH directly. The etiology of VH is complex with insights from preclinical and clinical studies suggesting both genetics and variation in gut microbiome composition play a role in susceptibility to IBS/VH. This project examines the bidirectional relationship between 1) the host-gene expression of a novel VH candidate gene, Avpr1a, encoding for arginine vasopressin receptor 1A, and 2) microbial colonization of the colon with the goal of identifying novel non- opioid, gastrointestinal tissue-specific, therapeutic targets to prevent/treat VH.

Methods

Two model BL/6 substrains C57BL/6NTac (Taconic) and C57BL/6J (Jackson), were used to compare differences in VH using an intracolonic instillation of ZYM, a protein-carbohydrate complex found in the cell walls of the yeast Saccharomyces cerevisiae. The use of ZYM results in a VH in the absence of structural damage or ongoing inflammation in the colon, which accurately parallels the clinical presentation of IBS. Leveraging whole genome sequencing of BL/6 substrains, we identified a list of candidate genes related to VH susceptibility. Our multi-omic approaches using mRNA/protein expression, calcium imaging, and microbiome analysis (16S rRNA sequencing) has allowed us to understand the individual contributions of gene-host expression and microbiota/microbiome to VH. Furthermore, manipulation of gene-host expression through an antisense oligonucleotide (ASO), and the manipulation of microbiota composition through fecal microbiota transplants (FMT), have also been explored.

Results

Two C57BL/6 mouse substrains, C57BL/6NTac (Taconic) and C57BL/6J (Jackson), have differential susceptibility to VH induced by intracolonic ZYM. C57BL/6NTac mice develop persistent VH while C57BL/6J mice are resistant. Using genomic comparisons, we have identified arginine-vasopressin receptor 1A (Avpr1a) as the highest priority candidate gene for VH susceptibility. Subsequent data also reveals an upregulation of colonic enteric neuron-specific expression of Avpr1a in C57BL/6NTac mice with VH. Our data also suggest a potential relationship between enteric neuron-specific expression of Avpr1a and the gut microenvironment, including increased colonization with Firmicutes Lachnospiracaea Dorea and impaired intestinal barrier permeability. Further, we’ve identified colon-specific alterations in enteric neuron response properties that covary with gene expression and the pattern of microbial colonization corresponding to VH development.

Conclusions

We propose Avpr1a as (1) a novel colon-specific therapeutic target for VH susceptibility for IBS and other disorder of gut-brain interactions. Using a low dose Avpr1a-specific ASO has further helped identify potential benefits of localized knockdown of Avpr1a to abolish VH, and importantly fill in the knowledge gap on the contribution of enteric neurons to visceral pain sensitization in patients. We have also developed a FMT protocol that decreases VH, highlight the innovation in our combination of studying both host physiology/genetics and the gut-microbiome. Further work will focus on whether microbiome changes results from or results in altered host-gene expression and VH. This study will shed light on new possibilities for the treatment and/or prevention of VH in DGBI patients by understanding the multifactorial relationship between the gut microbial colonization/function, Avpr1a expression, ENS neuronal responses, and VH susceptibility.

References

Below are the current references that I have used to support my work and written content of this submission. I did not add numerical references in each sections due to limited space.

1.Spiller R, Aziz Q, Creed F, Emmanuel A, Houghton L, Hungin P, et al. Guidelines on the irritable bowel syndrome: mechanisms and practical management. Gut. 2007;56(12):1770-98.
2.Hungin AP, Whorwell PJ, Tack J, Mearin F. The prevalence, patterns and impact of irritable bowel syndrome: an international survey of 40,000 subjects. Aliment Pharmacol Ther. 2003;17(5):643-50.
3.Mayer EA, Gebhart GF. Basic and clinical aspects of visceral hyperalgesia. Gastroenterology. 1994;107(1):271-93.
4.Camilleri M, Saslow SB, Bharucha AE. Gastrointestinal sensation. Mechanisms and relation to functional gastrointestinal disorders. Gastroenterol Clin North Am. 1996;25(1):247-58.
5.Camilleri M, Coulie B, Tack JF. Visceral hypersensitivity: facts, speculations, and challenges. Gut. 2001;48(1):125-31.
6.Barbara G, De Giorgio R, Stanghellini V, Cremon C, Salvioli B, Corinaldesi R. New pathophysiological mechanisms in irritable bowel syndrome. Aliment Pharmacol Ther. 2004;20 Suppl 2:1-9.
7.Basnayake C. Treatment of irritable bowel syndrome. Aust Prescr. 2018;41(5):145-9.
8.Sayuk GS, Kanuri N, Gyawali CP, Gott BM, Nix BD, Rosenheck RA. Opioid medication use in patients with gastrointestinal diagnoses vs unexplained gastrointestinal symptoms in the US Veterans Health Administration. Aliment Pharmacol Ther. 2018;47(6):784-91.
9.Camilleri M. Genetics of human gastrointestinal sensation. Neurogastroenterol Motil. 2013;25(6):458-66.
10.Drossman DA, Camilleri M, Mayer EA, Whitehead WE. AGA technical review on irritable bowel syndrome. Gastroenterology. 2002;123(6):2108-31.
11.Pimentel M, Lembo A. Microbiome and Its Role in Irritable Bowel Syndrome. Dig Dis Sci. 2020;65(3):829-39.
12.Wouters MM, Lambrechts D, Knapp M, Cleynen I, Whorwell P, Agreus L, et al. Genetic variants in CDC42 and NXPH1 as susceptibility factors for constipation and diarrhoea predominant irritable bowel syndrome. Gut. 2014;63(7):1103-11.
13.Saito YA, Petersen GM, Larson JJ, Atkinson EJ, Fridley BL, de Andrade M, et al. Familial aggregation of irritable bowel syndrome: a family case-control study. Am J Gastroenterol. 2010;105(4):833-41.
14.Saito YA, Petersen GM, Locke GR, 3rd, Talley NJ. The genetics of irritable bowel syndrome. Clin Gastroenterol Hepatol. 2005;3(11):1057-65.
15.Saito YA, Zimmerman JM, Harmsen WS, De Andrade M, Locke GR, 3rd, Petersen GM, et al. Irritable bowel syndrome aggregates strongly in families: a family-based case-control study. Neurogastroenterol Motil. 2008;20(7):790-7.
16.Tao ZY, Xue Y, Li JF, Traub RJ, Cao DY. Do MicroRNAs Modulate Visceral Pain? Biomed Res Int. 2018;2018:5406973.
17.Ren TH, Wu J, Yew D, Ziea E, Lao L, Leung WK, et al. Effects of neonatal maternal separation on neurochemical and sensory response to colonic distension in a rat model of irritable bowel syndrome. Am J Physiol Gastrointest Liver Physiol. 2007;292(3):G849-56.
18.Bradesi S, Lao L, McLean PG, Winchester WJ, Lee K, Hicks GA, et al. Dual role of 5-HT3 receptors in a rat model of delayed stress-induced visceral hyperalgesia. Pain. 2007;130(1-2):56-65.
19.Videlock EJ, Mahurkar-Joshi S, Hoffman JM, Iliopoulos D, Pothoulakis C, Mayer EA, et al. Sigmoid colon mucosal gene expression supports alterations of neuronal signaling in irritable bowel syndrome with constipation. Am J Physiol Gastrointest Liver Physiol. 2018;315(1):G140-G57.
20.Fuentes IM, Christianson JA. Ion channels, ion channel receptors, and visceral hypersensitivity in irritable bowel syndrome. Neurogastroenterol Motil. 2016;28(11):1613-8.
21.del Fresno C, Soler-Rangel L, Soares-Schanoski A, Gomez-Pina V, Gonzalez-Leon MC, Gomez-Garcia L, et al. Inflammatory responses associated with acute coronary syndrome up-regulate IRAK-M and induce endotoxin tolerance in circulating monocytes. Journal of endotoxin research. 2007;13(1):39-52.
22.Zhu S, Min L, Guo Q, Li H, Yu Y, Zong Y, et al. Transcriptome and methylome profiling in a rat model of irritable bowel syndrome induced by stress. Int J Mol Med. 2018;42(5):2641-9.
23.Mortazavi M, Ren Y, Saini S, Antaki D, Pierre CS, Williams A, et al. Polymorphic SNPs, short tandem repeats and structural variants are responsible for differential gene expression across C57BL/6 and C57BL/10 substrains. bioRxiv. 2021:2020.03.16.993683.
24.Peery AF, Crockett SD, Murphy CC, Jensen ET, Kim HP, Egberg MD, et al. Burden and Cost of Gastrointestinal, Liver, and Pancreatic Diseases in the United States: Update 2021. Gastroenterology. 2022;162(2):621-44.
25.Everhart JE, Ruhl CE. Burden of digestive diseases in the United States Part III: Liver, biliary tract, and pancreas. Gastroenterology. 2009;136(4):1134-44.
26.Inadomi JM, Fennerty MB, Bjorkman D. Systematic review: the economic impact of irritable bowel syndrome. Aliment Pharmacol Ther. 2003;18(7):671-82.
27.Hser YI, Mooney LJ, Saxon AJ, Miotto K, Bell DS, Huang D. Chronic pain among patients with opioid use disorder: Results from electronic health records data. J Subst Abuse Treat. 2017;77:26-30.
28.Farzaei MH, Bahramsoltani R, Abdollahi M, Rahimi R. The Role of Visceral Hypersensitivity in Irritable Bowel Syndrome: Pharmacological Targets and Novel Treatments. J Neurogastroenterol Motil. 2016;22(4):558-74.
29.Morris GP, Beck PL, Herridge MS, Depew WT, Szewczuk MR, Wallace JL. Hapten-induced model of chronic inflammation and ulceration in the rat colon. Gastroenterology. 1989;96(3):795-803.
30.Sengupta JN, Snider A, Su X, Gebhart GF. Effects of kappa opioids in the inflamed rat colon. Pain. 1999;79(2-3):175-85.
31.Lu G, Qian X, Berezin I, Telford GL, Huizinga JD, Sarna SK. Inflammation modulates in vitro colonic myoelectric and contractile activity and interstitial cells of Cajal. Am J Physiol. 1997;273(6):G1233-45.
32.Verma-Gandhu M, Verdu EF, Bercik P, Blennerhassett PA, Al-Mutawaly N, Ghia JE, et al. Visceral pain perception is determined by the duration of colitis and associated neuropeptide expression in the mouse. Gut. 2007;56(3):358-64.
33.Zhang HQ, Ding TT, Zhao JS, Yang X, Zhang HX, Zhang JJ, et al. Therapeutic effects of Clostridium butyricum on experimental colitis induced by oxazolone in rats. World J Gastroenterol. 2009;15(15):1821-8.
34.Burton MB, Gebhart GF. Effects of intracolonic acetic acid on responses to colorectal distension in the rat. Brain Res. 1995;672(1-2):77-82.
35.Kamp EH, Jones RC, 3rd, Tillman SR, Gebhart GF. Quantitative assessment and characterization of visceral nociception and hyperalgesia in mice. Am J Physiol Gastrointest Liver Physiol. 2003;284(3):G434-44.
36.Canavan C, West J, Card T. The epidemiology of irritable bowel syndrome. Clin Epidemiol. 2014;6:71-80.
37.Christianson JA, Gebhart GF. Assessment of colon sensitivity by luminal distension in mice. Nat Protoc. 2007;2(10):2624-31.
38.Feng B, Gebhart GF. Characterization of silent afferents in the pelvic and splanchnic innervations of the mouse colorectum. Am J Physiol Gastrointest Liver Physiol. 2011;300(1):G170-80.
39.Feng B, La JH, Schwartz ES, Tanaka T, McMurray TP, Gebhart GF. Long-term sensitization of mechanosensitive and -insensitive afferents in mice with persistent colorectal hypersensitivity. Am J Physiol Gastrointest Liver Physiol. 2012;302(7):G676-83.
40.Shinoda M, Feng B, Gebhart GF. Peripheral and central P2X receptor contributions to colon mechanosensitivity and hypersensitivity in the mouse. Gastroenterology. 2009;137(6):2096-104.
41.Zurita E, Chagoyen M, Cantero M, Alonso R, Gonzalez-Neira A, Lopez-Jimenez A, et al. Genetic polymorphisms among C57BL/6 mouse inbred strains. Transgenic Res. 2011;20(3):481-9.
42.Wilson SG, Chesler EJ, Hain H, Rankin AJ, Schwarz JZ, Call SB, et al. Identification of quantitative trait loci for chemical/inflammatory nociception in mice. Pain. 2002;96(3):385-91.
43.Nair HK, Hain H, Quock RM, Philip VM, Chesler EJ, Belknap JK, et al. Genomic loci and candidate genes underlying inflammatory nociception. Pain. 2011;152(3):599-606.
44.Mogil JS, Sorge RE, LaCroix-Fralish ML, Smith SB, Fortin, A., Sotocinal SG, Ritchie, J., Austin JS, et al. Pain sensitivity and vasopressin analgesia are mediated by a gene-sex-environment interaction. Nat Neuroscie. 2011;14(12):1569-73.
45.Roach KL, Hershberger PE, Rutherford JN, Molokie RE, Wang ZJ, Wilkie DJ. The AVPR1A Gene and Its Single Nucleotide Polymorphism rs10877969: A Literature Review of Associations with Health Conditions and Pain. Pain management nursing : official journal of the American Society of Pain Management Nurses. 2018;19(4):430-44.
46.Nishina K, Takagishi H, Takahashi H, Sakagami M, Inoue-Murayama M. Association of Polymorphism of Arginine-Vasopressin Receptor 1A (AVPR1a) Gene With Trust and Reciprocity. Front Hum Neurosci. 2019;13:230.
47.Christianson JA, Bielefeldt K, Altier C, Cenac N, Davis BM, Gebhart GF, et al. Development, plasticity and modulation of visceral afferents. Brain research reviews. 2009;60(1):171-86.
48.Jones RC, 3rd, Xu L, Gebhart GF. The mechanosensitivity of mouse colon afferent fibers and their sensitization by inflammatory mediators require transient receptor potential vanilloid 1 and acid-sensing ion channel 3. J Neurosci. 2005;25(47):10981-9.
49.Jones RC, 3rd, Otsuka E, Wagstrom E, Jensen CS, Price MP, Gebhart GF. Short-term sensitization of colon mechanoreceptors is associated with long-term hypersensitivity to colon distention in the mouse. Gastroenterology. 2007;133(1):184-94.
50.Smith HS. Activated microglia in nociception. Pain physician.13(3):295-304.
51.Li HG, Zhou ZG, Li Y, Zheng XL, Lei S, Zhu L, et al. Alterations of Toll-like receptor 4 expression on peripheral blood monocytes during the early stage of human acute pancreatitis. Dig Dis Sci. 2007;52(8):1973-8.
52.Chiu IM, Heesters BA, Ghasemlou N, Von Hehn CA, Zhao F, Tran J, et al. Bacteria activate sensory neurons that modulate pain and inflammation. Nature. 2013;501(7465):52-7.
53.Ochoa-Cortes F, Ramos-Lomas T, Miranda-Morales M, Spreadbury I, Ibeakanma C, Barajas-Lopez C, et al. Bacterial cell products signal to mouse colonic nociceptive dorsal root ganglia neurons. Am J Physiol Gastrointest Liver Physiol. 2010;299(3):G723-32.
54.Acosta C, Davies A. Bacterial lipopolysaccharide regulates nociceptin expression in sensory neurons. Journal of neuroscience research. 2008;86(5):1077-86.
55.Saito S, Okuno A, Cao DY, Peng Z, Wu HY, Lin SH. Bacterial Lipoteichoic Acid Attenuates Toll-Like Receptor Dependent Dendritic Cells Activation and Inflammatory Response. Pathogens. 2020;9(10).
56.Ashida K, Miyazaki K, Takayama E, Tsujimoto H, Ayaori M, Yakushiji T, et al. Characterization of the expression of TLR2 (toll-like receptor 2) and TLR4 on circulating monocytes in coronary artery disease. Journal of atherosclerosis and thrombosis. 2005;12(1):53-60.
57.Buchholz BM, Billiar TR, Bauer AJ. Dominant role of the MyD88-dependent signaling pathway in mediating early endotoxin-induced murine ileus. Am J Physiol Gastrointest Liver Physiol. 2010;299(2):G531-8.
58.Hironaka K, Yamaguchi Y, Okita R, Okawaki M, Nagamine I. Essential requirement of toll-like receptor 4 expression on CD11c+ cells for locoregional immunotherapy of malignant ascites using a streptococcal preparation OK-432. Anticancer research. 2006;26(5B):3701-7.
59.Hutchinson MR, Zhang Y, Shridhar M, Evans JH, Buchanan MM, Zhao TX, et al. Evidence that opioids may have toll-like receptor 4 and MD-2 effects. Brain Behav Immun. 2010;24(1):83-95.
60.Methe H, Kim JO, Kofler S, Weis M, Nabauer M, Koglin J. Expansion of circulating Toll-like receptor 4-positive monocytes in patients with acute coronary syndrome. Circulation. 2005;111(20):2654-61.
61.Peri F, Piazza M, Calabrese V, Damore G, Cighetti R. Exploring the LPS/TLR4 signal pathway with small molecules. Biochem Soc Trans.38(5):1390-5.
62.Fukushima R, Soejima H, Fukunaga T, Nakayama M, Oe Y, Oshima S, et al. Expression levels of Toll-like receptor genes in coronary atherosclerotic lesions of patients with acute coronary syndrome or stable angina pectoris. Circ J. 2009;73(8):1479-84.
63.Kanuri G, Ladurner R, Skibovskaya J, Spruss A, Konigsrainer A, Bischoff SC, et al. Expression of toll-like receptors 1-5 but not TLR 6-10 is elevated in livers of patients with non-alcoholic fatty liver disease. Liver Int. 2015;35(2):562-8.
64.Koch A, Hamann L, Schott M, Boehm O, Grotemeyer D, Kurt M, et al. Genetic variation of TLR4 influences immunoendocrine stress response: an observational study in cardiac surgical patients. Critical care (London, England).15(2):R109.
65.Bettoni I, Comelli F, Rossini C, Granucci F, Giagnoni G, Peri F, et al. Glial TLR4 receptor as new target to treat neuropathic pain: efficacy of a new receptor antagonist in a model of peripheral nerve injury in mice. Glia. 2008;56(12):1312-9.
66.Spiller R, Aziz Q, Creed F, Emmanuel A, Houghton L, Hungin P, et al. Guidelines on the irritable bowel syndrome: mechanisms and practical management. Gut. 2007;56(12):1770-98.
67.Hungin AP, Whorwell PJ, Tack J, Mearin F. The prevalence, patterns and impact of irritable bowel syndrome: an international survey of 40,000 subjects. Aliment Pharmacol Ther. 2003;17(5):643-50.
68.Mayer EA, Gebhart GF. Basic and clinical aspects of visceral hyperalgesia. Gastroenterology. 1994;107(1):271-93.
69.Camilleri M, Saslow SB, Bharucha AE. Gastrointestinal sensation. Mechanisms and relation to functional gastrointestinal disorders. Gastroenterol Clin North Am. 1996;25(1):247-58.
5.Camilleri M, Coulie B, Tack JF. Visceral hypersensitivity: facts, speculations, and challenges. Gut. 2001;48(1):125-31.
70.Barbara G, De Giorgio R, Stanghellini V, Cremon C, Salvioli B, Corinaldesi R. New pathophysiological mechanisms in irritable bowel syndrome. Aliment Pharmacol Ther. 2004;20 Suppl 2:1-9.

Presenting Author

Leena Kader

Poster Authors

Leena Kader

BSc

University of Kansas Medical Center

Lead Author

E-mail

Topics

  • Specific Pain Conditions/Pain in Specific Populations: Visceral Pain – Gastrointestinal/Abdominal