Anandamide down-regulates placental transporter expression through CB2 receptor-mediated inhibition of cAMP synthesis

John T. Szilagyi, Gabriella M. Composto-Wahler, Laurie B. Joseph, Bingbing Wang, Todd Rosen, Jeffrey Laskin, Lauren Aleksunes

Research output: Contribution to journalArticle

Abstract

The BCRP/ABCG2 efflux transporter is expressed on the membrane of placental syncytiotrophoblasts and protects the fetus from toxicant exposure. Syncytiotrophoblasts arise from the fusion of cytotrophoblasts, a process negatively regulated by the endocannabinoid, anandamide (AEA). It is unknown whether AEA can influence fetal concentrations of xenobiotics by modulating the expression of transporters in syncytiotrophoblasts. Here, we sought to characterize and identify the mechanism(s) responsible for AEA-mediated down-regulation of the BCRP transporter in human placental explants and BeWo trophoblasts. Treatment of human placental explants with AEA (1 μM, 24 h) reduced hCGα syncytin-1, and BCRP mRNAs by ˜30%. Similarly, treatment of BeWo trophoblasts with AEA (0–10 μM, 3-24 h) coordinately down-regulated mRNAs for hCGß syncytin-2, and BCRP. In turn, AEA increased the sensitivity of trophoblasts to the cytotoxicity of mitoxantrone, a known BCRP substrate, and environmental and dietary contaminants including mycoestrogens and perfluorinated chemicals. AEA-treated trophoblasts also demonstrated reduced BCRP transport of the mycoestrogen zearalenone and the diabetes drug glyburide, labeled with BODIPY. The AEA-mediated reduction of BCRP mRNA was abrogated when placental cells were co-treated with AM630, a CB2 receptor inhibitor, or 8-Br-cAMP, a cAMP analog. AEA reduced intracellular cAMP levels in trophoblasts by 75% at 1 h, and completely inhibited forskolin-induced phosphorylation of the cAMP response element binding protein (CREB). AEA also decreased p-CREB binding to the BCRP promoter. Taken together, our data indicate that AEA down-regulates placental transporter expression and activity via CB2-cAMP signaling. This novel mechanism may explain the repression of placental BCRP expression observed during diseases of pregnancy.

Original languageEnglish (US)
Pages (from-to)331-342
Number of pages12
JournalPharmacological Research
Volume141
DOIs
StatePublished - Mar 1 2019

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Cannabinoid Receptor CB2
Trophoblasts
Down-Regulation
Cyclic AMP Response Element-Binding Protein
Messenger RNA
Zearalenone
anandamide
Mitoxantrone
Endocannabinoids
Glyburide
Xenobiotics
Colforsin
Protein Binding
Fetus
Phosphorylation

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

@article{95fe8c954bd843aabb2109b9cbea772f,
title = "Anandamide down-regulates placental transporter expression through CB2 receptor-mediated inhibition of cAMP synthesis",
abstract = "The BCRP/ABCG2 efflux transporter is expressed on the membrane of placental syncytiotrophoblasts and protects the fetus from toxicant exposure. Syncytiotrophoblasts arise from the fusion of cytotrophoblasts, a process negatively regulated by the endocannabinoid, anandamide (AEA). It is unknown whether AEA can influence fetal concentrations of xenobiotics by modulating the expression of transporters in syncytiotrophoblasts. Here, we sought to characterize and identify the mechanism(s) responsible for AEA-mediated down-regulation of the BCRP transporter in human placental explants and BeWo trophoblasts. Treatment of human placental explants with AEA (1 μM, 24 h) reduced hCGα syncytin-1, and BCRP mRNAs by ˜30{\%}. Similarly, treatment of BeWo trophoblasts with AEA (0–10 μM, 3-24 h) coordinately down-regulated mRNAs for hCG{\ss} syncytin-2, and BCRP. In turn, AEA increased the sensitivity of trophoblasts to the cytotoxicity of mitoxantrone, a known BCRP substrate, and environmental and dietary contaminants including mycoestrogens and perfluorinated chemicals. AEA-treated trophoblasts also demonstrated reduced BCRP transport of the mycoestrogen zearalenone and the diabetes drug glyburide, labeled with BODIPY. The AEA-mediated reduction of BCRP mRNA was abrogated when placental cells were co-treated with AM630, a CB2 receptor inhibitor, or 8-Br-cAMP, a cAMP analog. AEA reduced intracellular cAMP levels in trophoblasts by 75{\%} at 1 h, and completely inhibited forskolin-induced phosphorylation of the cAMP response element binding protein (CREB). AEA also decreased p-CREB binding to the BCRP promoter. Taken together, our data indicate that AEA down-regulates placental transporter expression and activity via CB2-cAMP signaling. This novel mechanism may explain the repression of placental BCRP expression observed during diseases of pregnancy.",
author = "Szilagyi, {John T.} and Composto-Wahler, {Gabriella M.} and Joseph, {Laurie B.} and Bingbing Wang and Todd Rosen and Jeffrey Laskin and Lauren Aleksunes",
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Anandamide down-regulates placental transporter expression through CB2 receptor-mediated inhibition of cAMP synthesis. / Szilagyi, John T.; Composto-Wahler, Gabriella M.; Joseph, Laurie B.; Wang, Bingbing; Rosen, Todd; Laskin, Jeffrey; Aleksunes, Lauren.

In: Pharmacological Research, Vol. 141, 01.03.2019, p. 331-342.

Research output: Contribution to journalArticle

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T1 - Anandamide down-regulates placental transporter expression through CB2 receptor-mediated inhibition of cAMP synthesis

AU - Szilagyi, John T.

AU - Composto-Wahler, Gabriella M.

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AU - Rosen, Todd

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