Linking medicinal cannabis to autotaxin–lysophosphatidic acid signaling

Abstract

Autotaxin is primarily known for the formation of lysophosphatidic acid (LPA) from lysophosphatidylcholine. LPA is an important signaling phospholipid that can bind to six G protein–coupled receptors (LPA1–6). The ATX-LPA signaling axis is a critical component in many physiological and pathophysiological conditions. Here, we describe a potent inhibition of Δ9-trans-tetrahydrocannabinol (THC), the main psychoactive compound of medicinal cannabis and related cannabinoids, on the catalysis of two isoforms of ATX with nanomolar apparent EC50values. Furthermore, we decipher the binding interface of ATX to THC, and its derivative 9(R)-Δ6a,10a-THC (6a10aTHC), by X-ray crystallography. Cellular experiments confirm this inhibitory effect, revealing a significant reduction of internalized LPA1in the presence of THC with simultaneous ATX and lysophosphatidylcholine stimulation. Our results establish a functional interaction of THC with autotaxin–LPA signaling and highlight novel aspects of medicinal cannabis therapy.

Linking the Autotaxin-LPA Axis to Medicinal Cannabis and the Endocannabinoid System

“…THC works as a partial inhibitor on the catalysis of both ATX-β and ATX-γ isoforms, as described previously by Eymery et al . [ 62 ]. The apparent EC 50 of THC with ATX-β and the substrate LPC 18:1 is 1025 ± 138 nM, with a magnitude of inhibition around 60%.…”

In-situserial crystallography facilitates 96-well plate structural analysis at low symmetry

“…Crystallisation was performed using the HTX platform at the EMBL Grenoble, in CrystalDirect (CD) plates. Crystals were obtained using an adapted protocol of the conditions previously described for r-ATX-beta (Eymery et al, 2023). The r-ATX-beta protein solution was concentrated to 3-4 mg/mL in Tris 50 mM, NaCl 150 mM at pH 8.…”

“…In this work, autotaxin from rat (r-ATX), which is a phosphodiesterase producing the lipid signalling molecule lysophosphatidic acid (LPA), has been used as a challenging case study. ATX is the main producer of LPA, forming the ATX-LPA signalling axis with LPA acting as a multifunctional lipid mediator by engagement with six dedicated G-protein coupled receptors (LPA1-6) (Eymery et al, 2023).…”

“…ATX regulates many pathophysiological processes e.g., vascular development, neuropathy pain, fibrosis, rheumatoid arthritis, sclerosis, and cancer . The X-ray structure of r-ATX (MW 100 kDa) has already been determined through conventional cryo-MX (Hausmann et al, 2011;Eymery et al, 2023Eymery et al, , 2024, giving a good control over crystallisation, which was minimally optimised to obtain sitting drops containing micro-crystals. r-ATX was expressed in a HEK293T cellline, producing ~4 mg/mL purified protein (Eymery et al, 2023), which is inadequate for batch crystallisation.…”

“…The X-ray structure of r-ATX (MW 100 kDa) has already been determined through conventional cryo-MX (Hausmann et al, 2011;Eymery et al, 2023Eymery et al, , 2024, giving a good control over crystallisation, which was minimally optimised to obtain sitting drops containing micro-crystals. r-ATX was expressed in a HEK293T cellline, producing ~4 mg/mL purified protein (Eymery et al, 2023), which is inadequate for batch crystallisation. Thus, r-ATX serves as a relevant real-life case, justifying the viability of a crystallisation plate as the most natural sample delivery approach for serial crystallography experiments.…”