Dataset Information

Development of a Cannabinoid-Based Photoaffinity Probe to Determine the ?^8/9-Tetrahydrocannabinol Protein Interaction Landscape in Neuroblastoma Cells.

ABSTRACT: Introduction: ?⁹-Tetrahydrocannabinol (THC), the principle psychoactive ingredient in Cannabis, is widely used for its therapeutic effects in a large variety of diseases, but it also has numerous neurological side effects. The cannabinoid receptors (CBRs) are responsible to a large extent for these, but not all biological responses are mediated via the CBRs. Objectives: The identification of additional target proteins of THC to enable a better understanding of the (adverse) physiological effects of THC. Methods: In this study, a chemical proteomics approach using a two-step photoaffinity probe is applied to identify potential proteins that may interact with THC. Results: Photoaffinity probe 1, containing a diazirine as a photocrosslinker, and a terminal alkyne as a ligation handle, was synthesized in 14 steps. It demonstrated high affinity for both CBRs. Subsequently, two-step photoaffinity labeling in neuroblastoma cells led to identification of four potential novel protein targets of THC. The identification of these putative protein hits is a first step towards a better understanding of the protein interaction profile of THC, which could ultimately lead to the development of novel therapeutics based on THC.

SUBMITTER: Soethoudt M

PROVIDER: S-EPMC6038054 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Publications

Development of a Cannabinoid-Based Photoaffinity Probe to Determine the Δ8/9-Tetrahydrocannabinol Protein Interaction Landscape in Neuroblastoma Cells.

Soethoudt Marjolein M Alachouzos Georgios G van Rooden Eva J EJ Moya-Garzón María Dolores MD van den Berg Richard J B H N RJBHN Heitman Laura H LH van der Stelt Mario M

Cannabis and cannabinoid research 20180701 1

Introduction: Δ9-Tetrahydrocannabinol (THC), the principle psychoactive ingredient in Cannabis, is widely used for its therapeutic effects in a large variety of diseases, but it also has numerous neurological side effects. The cannabinoid receptors (CBRs) are responsible to a large extent for these, but not all biological responses are mediated via the CBRs. Objectives: The identification of additional target proteins of THC to enable a better understanding of the ...[more]

PMID: 29992186

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Project description:CD8+ T cells can contribute to neuroinflammation by secretion of inflammatory cytokines like interferon γ (IFNγ) and tumor necrosis factor α (TNFα). Astrocytes, a glial cell in the brain, can be stimulated by IFNγ and TNFα to secrete the inflammatory cytokines, monocyte chemotactic protein 1 (MCP-1), interleukin 6 (IL-6), and interferon-γ inducible protein 10 (IP-10). Δ9-Tetrahydrocannabinol (THC), the primary psychoactive cannabinoid in Cannabis sativa, possesses potent anti-inflammatory activity. The objective of this investigation was to assess the effects of THC treatment on CD8+ T cell-mediated activation of astrocytes. CD3/CD28/IFNα- stimulated CD8+ T cells were treated with vehicle (0.03% EtOH) or THC and cocultured with U251 astrocytes. IP-10+, MCP-1+, and IL-6+ astrocytes were quantified by flow cytometry. LegendPlex™ was used to measure cytokine secretion by CD8+ T cells and flow cytometry was employed to quantify IFNγ, TNFα, and lysosomal-associated membrane protein 1 (LAMP-1) expression. Recombinant TNFα and IFNγ were used to stimulate MCP-1, IP-10, IL-6 responses in U251 astrocytes, which were measured by flow cytometry. Treatment with THC reduced CD8+ T cell-mediated induction of IP-10 and IL-6 responses in U251 astrocytes but had no effect on MCP-1. THC treatment differentially affected T cell effector functions such that IFNγ and degranulation responses were sensitive to THC-mediated ablation while TNFα was not. Lastly, THC treatment reduced the IFNγ-induced IP-10 response but had no effect on TNFα-induced MCP-1 response in U251 astrocytes. The results suggest that cannabinoid treatment can selectively reduce certain CD8+ T cell responses that contribute to stimulation of astrocytes. Graphical Abstract Treatment with THC can abate CD8+ T cell-dependent neuroinflammatory processes by inhibiting CD8+ cell differentiation into effector cells, suppressing CD8+ effector cell function, and reducing activation of astrocytes by CD8+ T cell-derived inflammatory cytokines.

Dataset Information

Development of a Cannabinoid-Based Photoaffinity Probe to Determine the ?^8/9-Tetrahydrocannabinol Protein Interaction Landscape in Neuroblastoma Cells.

Publications

Development of a Cannabinoid-Based Photoaffinity Probe to Determine the Δ<sup>8/9</sup>-Tetrahydrocannabinol Protein Interaction Landscape in Neuroblastoma Cells.

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Dataset Information

Development of a Cannabinoid-Based Photoaffinity Probe to Determine the ?8/9-Tetrahydrocannabinol Protein Interaction Landscape in Neuroblastoma Cells.

Publications

Development of a Cannabinoid-Based Photoaffinity Probe to Determine the Δ<sup>8/9</sup>-Tetrahydrocannabinol Protein Interaction Landscape in Neuroblastoma Cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Development of a Cannabinoid-Based Photoaffinity Probe to Determine the ?^8/9-Tetrahydrocannabinol Protein Interaction Landscape in Neuroblastoma Cells.