Cannabis, in addition to its psychotropic properties, could have anti-inflammatory and immunomodulatory effects. Phytocannabinoids (pCBs) are a group of molecules naturally secreted by the cannabis plant. The major pCBs are cannabidiol (CBD) and Δ9-tetrahydrocannabinol Δ9 (THC). Only THC has psychotropic effects, which CBD does not have. Alongside these two main components, there is a wide variety of other molecules, such as other pCBs and terpenes which could increase the effects on immune system through synergistic interactions between these different compounds ("entourage effect").In vivo, pCBs essentially interfere with the endocannabinoid system, acting on many ubiquitous receptors, present on a significant number of different cell types. Numerous published studies show that pCBs have immunomodulatory and anti-inflammatory properties by acting on several of these receptors, whether through modulation of the immune response of different cell types, effects on cytokine networks, reduction of innate and adaptive responses and/or impact on cell survival or death (autophagy, proliferation/ apoptosis). The Immune-Mediated Inflammatory Diseases (IMIDs) affect 5 to 7% of the general population in Western countries, involve different organs (joints, skin, digestive tract) but share the same inflammatory mechanisms resulting from a dysregulation of the immune response. Our research focuses on the identification of the most effective phytochemical profile of pCBs, allowing an optimal effect on chronic inflammatory pathologies of interest among immune-mediated chronic inflammatory diseases (IMIDs). The pCB-IMIDs project is therefore part of an innovative translational project, around new therapeutic applications of medical cannabis (CannAppIMIDs). In our study, we will include 100 patients with one of IMIDs among Rheumatoid Arthritis, spondylarthritis, psoriatic arthritis, Sjogren disease and systemic lupus, at different stage and with different treatments. After patient's consent we will collect for research purposes an additional 40 ml of blood during a routine care blood test. Mononuclear and polynucleated blood cells will be exposed in vitro to different full-spectrum pCB extracts (full spectrum extract) including a CBD dominant and low THC extract (\<0.2%), 1 dominant THC extract, 1 balanced THC/CBD extract and 1 dominant CBG extract. In this cross-sectional study, our objective will be to assess the biological effects of different pCB compositions on inflammatory profiles (concentrations of pro and anti-inflammatory cytokines and chemokines) and modulations of expression profiles (autophagy, apoptosis, and cannabinoid receptor expression profile).
Male or female ≥ 18 years old
Diagnosis confirmed by a rheumatologist of RA or spondyloarthropathy (with or without IBD) or psoriatic arthritis (with or without active psoriasis) or systemic lupus erythematosus or Sjögren's disease
Patient who has expressed consent to participate in the study
Patients affiliated to social security
Patient who received intravenous corticosteroid therapy less than 4 weeks ago
Patient receiving oral corticosteroid therapy with a daily dose >15 mg/day
Consumption of CBD and/or recreational cannabis and/or positive saliva test for cannabis consumption and/or CBD
Pregnant and lactating women
In this exploratory study, the study population will include subjects suffering from IMID with rheumatological expression: rheumatoid arthritis (RA), Spondyloarthropathies with or without IBD, Psoriatic arthritis (Rpso), systemic lupus erythematosus (SLE), Sjögren's disease.
Other
Cross-sectional
1 blood sample, 40 mL (collection during a blood test, part of the standard of care for this disease)
Other interventionMononuclear and polynuclear peripheral blood cells from subjects suffering from IMID will be exposed in vitro to different inflammatory inducers with and without the concomitant addition of at least 4 different pCB extracts spectrum complete (full spectrum) including a dominant CBD extract and low in THC (\<0.2%), 1 dominant THC extract, 1 balanced THC/CBD extract and 1 dominant CBG extract. The same analyzes will be carried out in the presence of a recognized anti-inflammatory (for example hydrocortisone or dexamethasone) which will thus be used as a positive control.
The primary outcome is to evaluate the biological effect of exposure of blood cells from patients suffering from IMID to different varieties of pCB extracts on the inflammatory profile.
The initial inflammatory profiles of patients will be established by evaluations at the transcriptomic and protein level.
The primary outcome is to evaluate the immunological effect of exposure of blood cells from patients suffering from IMID to different varieties of pCB extracts on the inflammatory profile.
In plasma, the concentrations of cytokines and chemokines associated with inflammation, pro (mainly TNF-α IFN-α, but for instance also IL6, IL8, and IL1-β) and anti-inflammatory (for example IL-10) will be quantified in Multiplexed (34-plex type) or conventional ELISA. The level of expression of the corresponding transcripts, produced by the sorted subpopulations (cellular targets: polymorphonuclear, CD14+ monocyte, CD19+ B lymphocytes, CD4+ T4 lymphocytes and CD8+ T8 lymphocytes) will subsequently be reported by BRB-Seq \[13\] and/or by droplet RT-PCR (RT-ddPCR) assays. This quantification technique is reproducible, requires little starting biological material and is very efficient in detecting small quantities.
Inflammatory and autophagic/apoptotic transcripts, as well as transcripts of the main phytocannabinoid receptors in subpopulations of sorted blood cells.
Quantify ex vivo (by BRB-Seq and/or droplet quantitative RT-PCR (RT-ddPCR)). nflammatory and autophagic/apoptotic transcripts, as well as transcripts of the main phytocannabinoid receptors in subpopulations of sorted blood cells (polymorphonuclear cells, CD14+ monocytes, CD19+ B lymphocytes, CD4+ T and CD8+ T cells and if possible, quantify the evolution, after induction of an inflammatory context, of transcripts.
The main phytocannabinoid receptors in blood cells
Quantify ex vivo (by BRB-Seq and/or droplet quantitative RT-PCR (RT-ddPCR)). the expression profile of phyto and endocannabinoid receptors in the subpopulations will be established. Note that the development of this quantification of the transcripts of certain cytokines and several pCB receptors has already been carried out during previous work in the CBM laboratory (on French blood institution blood donors). Finally, an evaluation of mRNAs associated with apoptosis and autophagy could also be documented. Indeed, autophagy, a crucial biological function for the cell, linked to inflammation \[14, 15\], will also be evaluated, because the combined effects of pCB, pro-apoptotic, pro-autophagic and anti-inflammatory, have already been reported.
The main autophagy genes and apoptosis genes such as caspases
Quantify ex vivo (by BRB-Seq and/or droplet quantitative RT-PCR (RT-ddPCR)). Inflammatory and autophagic/apoptotic transcripts, as well as transcripts of the main phytocannabinoid receptors in subpopulations of sorted blood cells (polymorphonuclear cells, CD14+ monocytes, CD19+ B lymphocytes, CD4+ T and CD8+ T cells, and quantify the evolution, after induction of an inflammatory context, of transcripts.
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