Association Between a Polymorphism in Cannabinoid Receptor 2 and Severe Necroinflammation in Patients With Chronic Hepatitis C

doi:10.1016/j.cgh.2013.05.008

Clinical Gastroenterology and Hepatology

Volume 12, Issue 2, February 2014, Pages 334-340

https://doi.org/10.1016/j.cgh.2013.05.008 Get rights and content

Background & Aims

The cannabinoid receptor 2 (CB2) has been implicated in liver disease. The single-nucleotide polymorphism rs35761398 in cannabinoid receptor 2 gene (CNR2), which encodes the CB2, substitutes glutamine (Q) 63 with arginine (R), and reduces the function of the gene product. We investigated the effects of CNR2 rs35761398 in patients with hepatitis C virus (HCV) infection.

Methods

We studied 169 consecutive patients with asymptomatic chronic hepatitis (tested positive for anti-HCV and HCV RNA) at 2 liver units in southern Italy. First, liver biopsy samples were collected from July 2009 through December 2011. All patients were naive to antiviral therapy; CNR2 genotype was determined by polymerase chain reaction analysis.

Results

Patients with the CB2-63 QQ variant had higher serum levels of aminotransferase than those with the CB2-63 QR or RR variants; they also had higher histologic activity index (HAI) scores (8.6 ± 3.8) than patients without the CB2-63 RR variant (5.3 ± 3.6; P < .005) or those with the CB2-63 QR variant (5.8 ± 3.3; P < .001). Patients with the different variants of CNR2 did not differ in fibrosis stage or steatosis score. Moderate or severe chronic hepatitis (HAI score, >8) was identified more frequently (55.5%) in patients with the CB2-63 QQ variant than in those with the 63 QR (20%; P < .005) or RR variants (17.4%; P < .005). In logistic regression analysis, the CB2-63 QQ variant and fibrosis score were independent predictors of moderate or severe chronic hepatitis (HAI score, >8; P < .0001).

Conclusions

The CB2-63 QQ variant of CNR2 is associated with more severe inflammation and hepatocellular necrosis in patients with HCV infection.

Section snippets

Patients and Methods

Two Liver Units in the Campania region (southern Italy) participated in the study. These 2 centers have cooperated in several clinical investigations using the same clinical approach and the same laboratory methods.26, 27

A total of 169 consecutive Italian patients with anti-HCV/HCV RNA–positive asymptomatic chronic hepatitis for 18–36 months were included in the study. The patients enrolled were naive for antiviral therapy at the time they underwent their first liver biopsy from July 2009 to

Results

A total of 119 (70.4%) of the 169 patients had HCV genotype 1. The prevalence of patients with the CB2-63 QQ, QR, and RR variants was 10.7%, 62.1%, and 27.2%, respectively (Table 1), which is similar to previously reported distributions.30, 31, 32

The demographic, biochemical, virologic, and histologic data according to the CB2-63 variants are shown in Table 1. The mean aminotransferase serum level was higher in patients with the CB2-63 QQ variant (AST: mean ± SD, 2.6 ± 1.8 × normal value; ALT:

Discussion

This study analyzed the role of a functional polymorphism of CB type 2 in a cohort of 169 patients with CHC. The single-nucleotide polymorphism studied here is a missense mutation of the second and third base at codon 63 of the CNR2 gene, which leads to a Q/R substitution, causing a different polarization state of the protein. The CB2 variants have been shown to affect the ability of the CB2 receptor differently to exert its function.25, 33 The data from the present study suggest that the

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Cited by (49)

Immunomodulation by cannabinoids: Current uses, mechanisms, and identification of data gaps to be addressed for additional therapeutic application
2021, Advances in Pharmacology
Citation Excerpt :
As we have already noted, many autoimmune diseases are associated with the Q63R variant in the CB2 receptor. Interestingly, in Italian cohorts, the CB2 63 QQ variant was associated with higher serum liver enzymes and histologic activity index in those individuals with chronic hepatitis C (Coppola et al., 2014), while there was an association of the CB2 63 RR variant with fibrosis score and higher prevalence of hepatitis B infection (Coppola et al., 2015). Recall that the Q63R variant in the CB2 receptor resulted in impairment of endocannabinoid suppression of immune function (Sipe et al., 2005).
The endocannabinoid system plays a critical role in immunity and therefore its components, including cannabinoid receptors 1 and 2 (CB₁ and CB₂), are putative druggable targets for immune-mediated diseases. Whether modulating endogenous cannabinoid levels or interacting with CB₁ or CB₂ receptors directly, cannabinoids or cannabinoid-based therapeutics (CBTs) show promise as anti-inflammatory or immune suppressive agents. Herein we provide an overview of cannabinoid effects in animals and humans that provide support for the use of CBTs in immune-mediated disease such as multiple sclerosis (MS), inflammatory bowel disease (IBD), asthma, arthritis, diabetes, human immunodeficiency virus (HIV), and HIV-associated neurocognitive disorder (HAND). This is not an exhaustive review of cannabinoid effects on immune responses, but rather provides: (1) key studies in which initial and/or novel observations were made in animal studies; (2) critical human studies including meta-analyses and randomized clinical trials (RCTs) in which CBTs have been assessed; and (3) evidence for the role of CB₁ or CB₂ receptors in immune-mediated diseases through genetic analyses of single nucleotide polymorphisms (SNPs) in the CNR1 and CNR2 genes that encode CB₁ or CB₂ receptors, respectively. Perhaps most importantly, we provide our view of data gaps that exist, which if addressed, would allow for more rigorous evaluation of the efficacy and risk to benefit ratio of the use of cannabinoids and/or CBTs for immune-mediated diseases.
Hepatic stellate cells as key target in liver fibrosis
2017, Advanced Drug Delivery Reviews
Citation Excerpt :
Newer generation antagonists with improved target specificity and limited central nervous system penetration such as AM4113 and AM6545 have been developed based on the crystal structure of CB1 [167,168]. A germline genetic variant in the CNR2 gene (encoding CB2), CB2-63QQ, was found to be associated with more severe necroinflammation in patients with chronic hepatitis C [169]. A selective CB 2 agonist, JWH-133, reduces fibrosis by inducing HSC quiescence/apoptosis and reducing IL-17 production by Th17 cells [166,170].
Progressive liver fibrosis, induced by chronic viral and metabolic disorders, leads to more than one million deaths annually via development of cirrhosis, although no antifibrotic therapy has been approved to date. Transdifferentiation (or “activation”) of hepatic stellate cells is the major cellular source of matrix protein-secreting myofibroblasts, the major driver of liver fibrogenesis. Paracrine signals from injured epithelial cells, fibrotic tissue microenvironment, immune and systemic metabolic dysregulation, enteric dysbiosis, and hepatitis viral products can directly or indirectly induce stellate cell activation. Dysregulated intracellular signaling, epigenetic changes, and cellular stress response represent candidate targets to deactivate stellate cells by inducing reversion to inactivated state, cellular senescence, apoptosis, and/or clearance by immune cells. Cell type- and target-specific pharmacological intervention to therapeutically induce the deactivation will enable more effective and less toxic precision antifibrotic therapies.
Polymorphisms of the CB2 Cannabinoid Receptor
2017, Handbook of Cannabis and Related Pathologies: Biology, Pharmacology, Diagnosis, and Treatment
This chapter focuses on functional and genetic association studies of single nucleotide polymorphisms (SNPs) on human CB2 cannabinoid receptor (CNR2). A review of original papers, published in English and indexed in PubMed up to Jun., 2014, is discussed in this chapter. Nonsynonymous SNPs in CNR2 have been reported to cause changes of the responses of the CB2 to various cannabinoid ligands. In addition, nonsynonymous SNPs, synonymous SNPs, as well as SNPs in the noncoding region of CNR2 have been shown to be associated with a variety of diseases, including autoimmune diseases, liver diseases, osteoporosis, and disorders of central nervous system.
The endocannabinoid system in the baboon (Papio spp.) as a complex framework for developmental pharmacology
2016, Neurotoxicology and Teratology
Citation Excerpt :
The difference in the amino-acid sequences observed between baboons and humans is located in codon 63 (Q to R) (intracellular loop one). In humans, this codon is associated with polymorphism, and linked to auto-immune diseases (Sipe et al., 2005), depression (Onaivi et al., 2008), celiac disease (rs35761398), (Rossi et al., 2012), immune thrombocytopenic purpura (Mahmoud Gouda and Mohamed Kamel, 2013; Rossi et al., 2011; Rossi et al., 2012), hepatonecrosis (Coppola et al., 2014), and age of menarche (Bellini et al., 2015). Interestingly, the differences in the amino-acid sequence observed between humans and baboons in our study are mostly located in the C-terminus of CB2R (Fig. 4), which may be responsible for signal transduction (Dhopeshwarkar and Mackie, 2014) but is not critical for receptor binding (Feng et al., 2014).
The consumption of marijuana (exogenous cannabinoid) almost doubled in adults during last decade. Consumption of exogenous cannabinoids interferes with the endogenous cannabinoid (or “endocannabinoid” (eCB)) system (ECS), which comprises N-arachidonylethanolamide (anandamide, AEA), 2-arachidonoyl glycerol (2-AG), endocannabinoid receptors (cannabinoid receptors 1 and 2 (CB1R and CB2R), encoded by CNR1 and CNR2, respectively), and synthesizing/degrading enzymes (FAAH, fatty-acid amide hydrolase; MAGL, monoacylglycerol lipase; DAGL-α, diacylglycerol lipase-alpha). Reports regarding the toxic and therapeutic effects of pharmacological compounds targeting the ECS are sometimes contradictory. This may be caused by the fact that structure of the eCBs varies in the species studied.
First: to clone and characterize the cDNAs of selected members of ECS in a non-human primate (baboon, Papio spp.), and second: to compare those cDNA sequences to known human structural variants (single nucleotide polymorphisms and haplotypes).
Polymerase chain reaction-amplified gene products from baboon tissues were transformed into Escherichia coli. Amplicon-positive clones were sequenced, and the obtained sequences were conceptually translated into amino-acid sequences using the genetic code.
Among the ECS members, CNR1 was the best conserved gene between humans and baboons. The phenotypes associated with mutations in the untranslated regions of this gene in humans have not been described in baboons. One difference in the structure of CNR2 between humans and baboons was detected in the region with the only known clinically relevant polymorphism in a human receptor. All of the differences in the amino-acid structure of DAGL-α between humans and baboons were located in the hydroxylase domain, close to phosphorylation sites. None of the differences in the amino-acid structure of MAGL observed between baboons and humans were located in the area critical for enzyme function.
The evaluation of the data, obtained in non-human primate model of cannabis-related developmental exposure should take into consideration possible evolutionary-determined species-specific differences in the CB1R expression, CB2R transduction pathway, and FAAH and DAGLα substrate-enzyme interactions.
CB2-63 polymorphism and immune-mediated diseases associated with HCV chronic infection
2016, Digestive and Liver Disease
Citation Excerpt :
In accordance with this, we recently demonstrated the association of the CB2-63 RR variant with childhood immune thrombocytopenic purpura [40], celiac disease susceptibility [41] and the clinical course of juvenile arthritis [42]. Previously, we have observed that the CB2-63 QQ variant was associated with a more severe liver disease in a cohort of 169 patients with chronic hepatitis C [21]. Thus, in chronic hepatitis C the patients with QQ variants showed a more severe histological activity index and hepatocellular necrosis and those with RR variants a more frequent immune-mediated disorders, probably because the T cells with QQ variants were more inhibited with a less vigorous immune response against HCV, those with RR variants less inhibited, with a more vigorous systemic immune response.
To evaluate whether CB2 variants are associated with the presence of immune-mediated disorders (IMDs) in patients with chronic HCV infection.
One hundred and sixty-eight anti-HCV/HCV-RNA-positive patients were enrolled, 81 with signs of IMDs and 87 without. In the IMDs group, 22 (27.2%) showed ANA positivity (titers ≥1:160), 3 (3.7%) SMA positivity (titers ≥1:160), 24 (29.6%) had cryoglobulinemia, 25 (30.9%) autoimmune thyroiditis, 4 (4.9%) psoriasis, 2 (2.5%) B-cell non-Hodgkin lymphoma and 1 (1.2%) autoimmune hemolytic anemia. All patients were screened for the CNR2 rs35761398 single nucleotide polymorphism using a TaqMan Assay.
Compared with the 87 patients without IMDs, the 81 with IMDs were more frequently females (65% vs. 45%, p = 0.01), but no significant difference was found in the initial demographic, epidemiological, serological, biochemical or virological data. Instead, the prevalence of patients with the CB2-63 RR variant was significantly higher in the IMD than in the non-IMD group (49.4% vs. 24.1%, p = 0.001). A logistic regression analysis including the CB2-63 receptor (RR vs. QR or QQ), age and sex identified the CB2-63 RR as the only independent predictor of IMDs (p = 0.005).
The data suggest a significant, previously unknown, independent association between the CB2-63 RR variant and IMDs in anti-HCV-positive patients.
The study was approved by the Ethics Committee of the Azienda Ospedaliera Universitaria of the Second University of Naples (n° 214/2012).
The Role of Cannabidiol in Liver Disease: A Systemic Review
2024, International Journal of Molecular Sciences

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: Conflicts of interest The authors disclose no conflicts.

: Funding This study was supported by a grant from Progetti di Ricerca di Interesse Nazionale (PRIN) 2008, Ministero dell'Istruzione, dell'Università e della Ricerca Scientifica (MIUR), Rome, Italy “Ottimizzazione della Diagnosi Eziologica dell’Epatite Acuta C e Studio dei Fattori Viro-Immunologici di Guarigione, di Cronicizzazione e di Risposta Alla Terapia con Interferone,” and in part by a grant from Regione Campania “Progetti per il miglioramento della qualità dell'assistenza, diagnosi e terapia del paziente affetto da AIDS nei settori: immunologia, coinfezioni, informazione e prevenzione,” 2008.

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Original articlePancreas, biliary tract, and liverAssociation Between a Polymorphism in Cannabinoid Receptor 2 and Severe Necroinflammation in Patients With Chronic Hepatitis C

Background & Aims

Methods

Results

Conclusions

Section snippets

Patients and Methods

Results

Discussion

Gastroenterology

J Hepatol

Hepatology

J Clin Virol

Hepatology

Hepatology

J Hepatol

Clin Gastroenterol Hepatol

Clin Gastroenterol Hepatol

Gastroenterology

Hepatology

Addict Behav

J Biol Chem

J Hepatol

Pharmacol Res

Virology

Estimating future hepatitis C morbidity, mortality, and costs in the United States

Am J Public Health

Role of alcohol in the progression of liver disease caused by hepatitis C virus infection

Hepatology

Influence of alcohol use, race, and viral coinfections on spontaneous HCV clearance in a US veteran population

Hepatology

Daily cannabis smoking as a risk factor for progression of fibrosis in chronic hepatitis C

Hepatology

Genetic variation in IL28B and spontaneous clearance of hepatitis C virus

Nature

Original article
Pancreas, biliary tract, and liver
Association Between a Polymorphism in Cannabinoid Receptor 2 and Severe Necroinflammation in Patients With Chronic Hepatitis C