RT Journal Article
SR Electronic
T1 Multi-Omic Analysis Reveals Disruption of Cholesterol Homeostasis by Cannabidiol in Human Cell Lines
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.06.03.130864
DO 10.1101/2020.06.03.130864
A1 Steven E. Guard
A1 Douglas A. Chapnick
A1 Zachary C. Poss
A1 Christopher C. Ebmeier
A1 Jeremy Jacobsen
A1 Travis Nemkov
A1 Kerri A. Ball
A1 Kristofor J. Webb
A1 Helen L. Simpson
A1 Stephen Coleman
A1 Eric Bunker
A1 Adrian Ramirez
A1 Julie A. Reisz
A1 Robert Sievers
A1 Michael H.B. Stowell
A1 Angelo D’Alessandro
A1 Xuedong Liu
A1 William M. Old
YR 2021
UL http://biorxiv.org/content/early/2021/01/07/2020.06.03.130864.abstract
AB The non-psychoactive cannabinoid, cannabidiol (CBD), is FDA-approved for treatment of two drug-resistant epileptic disorders, and is seeing increased use among the general public, yet the mechanisms that underlie its therapeutic effects and side-effect profiles remain unclear. Here, we report a systems-level analysis of CBD action in human cell lines using temporal multi-omic profiling. FRET-based biosensor screening revealed that CBD treatment resulted in a sharp rise in cytosolic calcium, and activation of AMPK and ERK kinases in human keratinocyte and neuroblastoma cell lines. CBD treatment led to alterations in the abundance of metabolites, mRNA transcripts, and proteins consistent with activation of cholesterol biosynthesis, transport and storage. We found that CBD rapidly incorporated into cellular membranes and altered cholesterol chemical activity, suggesting direct perturbation of cholesterol-dependent membrane properties. CBD treatment induced apoptosis in a dose-dependent manner in multiple human cell lines, which was rescued by inhibition of cholesterol synthesis, and potentiated by compounds that disrupt cholesterol trafficking and storage. Our data point to a pharmacological interaction of CBD with cholesterol homeostasis pathways, with potential implications in its therapeutic use.Competing Interest StatementXL, DC and WO are patent holders of PCT WO2019246632A1, and XL, DC, WO and EB are patent holders of PCT WO2019118837A1. Both patents are related to this work. Unrelated to the contents of this manuscript, AD and TN are founders of Omix Technologies Inc, D.C. is the founder of Biooloomics, Inc, and RS is the founder of Sievers Infinity LLC.25-OHC25-hydroxycholesterolACACAacetyl-CoA carboxylaseACATacyl-coenzyme A cholesterol O-acyltransferaseAMPKAMP-activated protein kinaseAMKARAMP-activated protein kinase activity reporterAPOBapolipoprotein BAPOEapolipoprotein EATF4activating transcription factor 4CBDcannabidiolCAMKKßCa2+/ calmodulin-dependent protein kinase kinase ßCaV3voltage gated calcium channel 3CREBcAMP response element-binding proteinDMSOdimethylsulfoxideDHAdocosahexaenoic acidDHCR2424-dehydrocholesterol reductaseEEF2eukaryotic elongation factor 2EEF2Keukaryotic elongation factor 2 kinaseERKextracellular signal-regulated kinaseECARextracellular acidification rateERendoplasmic reticulumFCCPtrifluoromethoxy carbonylcyanide phenylhydrazoneFDAFood and Drug AdministrationFRAPfluorescence recovery after photobleachingFRETFörster resonance energy transferGPR55G protein-coupled receptor 55GRAMD1GRAM domain containing protein 1GTPguanosine triphosphateHK1Hexokinase 1HMGCR3-hydroxy-3-methylglutaryl-CoA reductaseHSP60heat shock protein 60LDLRlow density lipoprotein receptorLPElysophosphatidylethanolamineMBCDmethyl ß cyclodextrinMCMCMarkov chain Monte CarloNaV1.1sodium channel, voltage-gated, type I, alpha subunitNFE2L2nuclear factor erythroid 2-related factor 2NPC1Niemann-Pick disease, type C1OCRoxygen consumption ratePCAprincipal component analysisPCphosphatidylcholinePEphosphatidylethanolamineSCAPSREBP cleavage-activating proteinSGPL1Sphingosine-1-phosphate lyaseSILACstable isotope labeling of amino acids in cultureSOD1superoxide dismutase 1SP1specificity protein 1 transcription factorSREBPsterol regulatory element-binding proteinSTK11serine/ threonine kinase 11SUVsynthetic unilamellar vesicles TCA tricarboxylic acid cycleTMTtandem mass tagTRPM8transient receptor potential cation channel subfamily M member 8TRPVtransient receptor potentialVDAC1voltage-dependent anion channel 1