Differential expression and functional role of cannabinoid genes in alcohol users.
BACKGROUND
Genetic factors account for about fifty percent of the risk for alcoholism and alcohol dependence (AD) has been reported to be influenced by cannabinoid receptors (CBRs) and the endocannabinoid system (ECS). Previous studies have focused on cannabinoids and alcohol-related effects in the CNS; however, the role CBRs play on alcohol effects in the immune system has not been elucidated yet. Since alcohol can affect immune responses and have detrimental effects on immune cells such as dendritic cells (DCs), we hypothesize that alcohol can exert its effects on DCs by modulating changes in CBRs, which in turn can regulate important DC functions such as cytokine production.


METHODS
Therefore, we studied the expression of CNR1 and CNR2, and the novel cannabinoid G protein-coupled receptor (GPCR) 55 (GPR55) in human monocyte-derived dendritic cells (MDDCs) from alcohol users. CNR1, CNR2, and GPR55 genes were measured by qRT-PCR and protein by flow cytometry. MDDCs from alcohol users show significantly higher levels of CNR2 and GPR55 compared to MDDCs from non-users. These findings were further confirmed using MDDCs treated with alcohol. Inflammatory cytokines were measured in EtOH-treated and non-treated cells by antibody array.


RESULTS
Functional effects of CBRs on MDDCs were shown by CB2 and GPR55 siRNA transfection. Transfected EtOH-treated cells showed significantly higher levels of proinflammatory cytokine production as measured by IL-1β expression. Our results provide insights into alcohol mechanisms of DC regulation and show, for the first time, that alcohol is inducing CNR2 and GPR55 in human DCs.
Florida International University, United States. Electronic address: magudelo@fiu.edu.
Marisela
M
Agudelo
Adriana
A
Yndart
Marisol
M
Morrison
Gloria
G
Figueroa
Karla
K
Muñoz
Thangavel
T
Samikkannu
Madhavan P
MP
Nair
D000328:Adult; D000437:Alcoholism / Q000235:genetics*; D002186:Cannabinoids / Q000235:genetics*; D002454:Cell Differentiation / Q000187:drug effects; D002492:Central Nervous System Depressants / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000502:physiology; D000431:Ethanol / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D053583:Interleukin-1beta / Q000096:biosynthesis; D008297:Male; D008875:Middle Aged; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D043884:Receptor, Cannabinoid, CB1 / Q000235:genetics; D043885:Receptor, Cannabinoid, CB2 / Q000235:genetics*; D043882:Receptors, Cannabinoid / Q000235:genetics*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics; D014162:Transfection; D055815:Young Adult
D002186:Cannabinoids; D002492:Central Nervous System Depressants; D016207:Cytokines; C524885:GPR55 protein, human; D053583:Interleukin-1beta; D034741:RNA, Small Interfering; D043884:Receptor, Cannabinoid, CB1; D043885:Receptor, Cannabinoid, CB2; D043882:Receptors, Cannabinoid; D043562:Receptors, G-Protein-Coupled; D000431:Ethanol
Alcohol; Cannabinoid receptors; Cannabinoids; Human dendritic cells