Cannabidiol exerts antiinflammatory effects but maintains T effector memory cell differentiation in humans
Debora L. Gisch, Sachiko Koyama, Jumar Etkins, Gerald C. So, Daniel J. Fehrenbach, Jessica Bo Li Lu, Ying-Hua Cheng, Ricardo Melo Ferreira, Evan Rajadhyaksha, Kelsey McClara, Mahla Asghari, Asif A. Sharfuddin, Pierre C. Dagher, Laura M. Snell, Meena S. Madhur, Rafael B. Polidoro, Zeruesenay Desta, Michael T. Eadon
Debora L. Gisch, Sachiko Koyama, Jumar Etkins, Gerald C. So, Daniel J. Fehrenbach, Jessica Bo Li Lu, Ying-Hua Cheng, Ricardo Melo Ferreira, Evan Rajadhyaksha, Kelsey McClara, Mahla Asghari, Asif A. Sharfuddin, Pierre C. Dagher, Laura M. Snell, Meena S. Madhur, Rafael B. Polidoro, Zeruesenay Desta, Michael T. Eadon
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Clinical Research and Public Health Clinical Research Immunology

Cannabidiol exerts antiinflammatory effects but maintains T effector memory cell differentiation in humans

Abstract

BACKGROUND Cannabidiol (CBD) is increasingly used for pain management, including in transplant recipients with limited analgesic options. Its immunomodulatory effects in humans are not well defined at a single-cell level at CBD steady state with concomitant tacrolimus treatment.METHODS In a phase I ex vivo study, peripheral blood mononuclear cells from 23 participants who received oral CBD (Epidiolex) up to 5 mg/kg twice daily for 11 days were collected before CBD (pre-CBD) and at steady state (post-CBD). Lymphocytes were isolated and stimulated with anti-CD3/CD28 antibodies, with or without tacrolimus (5 ng/mL). Pharmacodynamic responses were assessed using CellTiter-Glo proliferation, single-cell and single-nucleus RNA sequencing, cytokine assays, and flow cytometry. Steady-state plasma concentrations of CBD were quantified via tandem mass spectrometry.RESULTS We identified an increased proportion of T effector memory (TEM) cells post-CBD (22% increase), which correlated with CBD plasma concentrations (R = 0.77, P = 0.01). CBD reduced proliferation of T (37% decrease) and CD70hi B (17% decrease) lymphocytes with additive immunosuppressive effects to tacrolimus. Single-cell RNA sequencing revealed reduced IL2 and TNF signaling and altered receptor-ligand networks in TEM cells. Post-CBD cytokine assays revealed elevated proinflammatory IL-6 protein levels and antiinflammatory IL-10 levels, with reduced TNF-α, LTA, and IL-2. In flow cytometry, the proportion of TEM and TEMRA cells increased post-CBD with tacrolimus.CONCLUSION CBD exerts mixed immunomodulatory effects in humans, combining antiproliferative and pro- and antiinflammatory responses. Understanding the clinical safety of CBD use is important given the paucity of pain control options available for immunocompromised transplant populations.TRIAL REGISTRATION ClinicalTrials.gov NCT05490511FUNDING NIH/National Center for Complementary and Integrative Health (R01AT011463); NIH/National Institute of General Medical Sciences (NIGMS) (R35GM145383); Intramural Research Program of the NIH; NIH/NIGMS (T32GM008425).

Authors

Debora L. Gisch, Sachiko Koyama, Jumar Etkins, Gerald C. So, Daniel J. Fehrenbach, Jessica Bo Li Lu, Ying-Hua Cheng, Ricardo Melo Ferreira, Evan Rajadhyaksha, Kelsey McClara, Mahla Asghari, Asif A. Sharfuddin, Pierre C. Dagher, Laura M. Snell, Meena S. Madhur, Rafael B. Polidoro, Zeruesenay Desta, Michael T. Eadon

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Figure 3

CBD concentration–dependent effects on cell proportion.

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CBD concentration–dependent effects on cell proportion. In the post-CBD ...
In the post-CBD phase, 10 participants had overlapping sc/snRNA-Seq and pharmacokinetic data. (A) Concentration-to-time curves of CBD concentration levels (ng/mL) at time points starting on day 12: 0 hours, 0.33 hours, 0.66 hours, 1 hour, 2 hours, 4 hours, 6 hours, and 12 hours while at CBD steady state. Some inter-individual variability was observed in the pharmacokinetic profiles of participants. (B) Scatterplot shows relationships between immune cell proportions and CBD concentrations in participants. Green dots represent proportion of TEM cells versus CBD trough concentration at steady state (C0h), with Pearson’s correlation coefficient (Cor) of 77%, P value of 0.01, and R2 value of 0.59. Dashed line indicates linear regression fits, performed using Pearson’s correlation by lm method in ggplot2 (v3.5.1). (C) Gene expression in TEM cells changed across comparisons: No CD3/CD28 –CBD vs. CD3/CD28 –CBD (x axis) and CD3/CD28 +CBD vs. CD3/CD28 –CBD (y axis). Scatterplot depicts log2 fold changes (log2FC) in gene expression for TEM cells in each comparison. Each point represents a gene with statistically significant differential expression (adjusted P value < 0.05) in both comparisons. Genes exhibiting opposing directions of regulation — up in one comparison and down in the other — are highlighted (green, upregulated with CBD under stimulation; gold, downregulated with CBD under stimulation). The top 300 genes identified based on discordant regulation across CBD treatment conditions and ranked by Euclidean distance from the origin (see Supplemental Table 6) were analyzed using active-subnetwork enrichment via pathfindR. Genes with adjusted P value less than 0.05 were input into pathfindR using the BioGRID protein-protein interaction network and KEGG gene set. (D) Volcano plot showing associations between CBD trough concentration (C0h) and pathway activity in TEM cells. Each point represents a pathway, with the x axis indicating effect size (correlation coefficient) and the y axis showing statistical significance (–log10 P value). N = 10 participants. (E–G) Negative correlations were observed for KEGG hsa03050 (proteasome, 90%, P = 0.00055; Pearson’s) and hsa00010 (glycolysis/gluconeogenesis, 80%, P = 0.0058; Pearson’s), while a positive correlation was identified for hsa00061 (fatty acid biosynthesis, 61%, P = 0.059; Pearson’s). For each participant in the x axis CBD trough C0h and y axis mean pathway activity (MPA) TEM cell divided by MPA calculated for all other cell types (CT).