Spatiotemporal regulation of cAMP signaling controls the human trophoblast fusion

Gerbaud, Pascale; Taskén, Kjetil; Pidoux, Guillaume

doi:10.3389/fphar.2015.00202

Cited by 34 publications

(28 citation statements)

References 82 publications

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“…Finally, AKAPs bind to or co-localize with specific PKA substrates to allow rapid and efficient phosphorylation [25]. Several AKAPs have been described in human placenta and we recently showed that two or more AKAPs are involved in the regulation of trophoblast fusion [20,[28][29][30]. Specifically, ezrin establishes a signaling complex with PKA and connexin 43 (Cx43) that mediates gap junction communication and thereby triggers trophoblast fusion [29,31].…”

Section: Introductionmentioning

confidence: 99%

Ezrin-anchored PKA phosphorylates serine 369 and 373 on connexin 43 to enhance gap junction assembly, communication, and cell fusion

Dukic

Gerbaud

Gadrey

et al. 2018

Biochemical Journal

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A limited number of human cells can fuse to form multinucleated syncytia. In the differentiation of human placenta, mononuclear cytotrophoblasts fuse to form an endocrinologically active, non-proliferative, multinucleated syncytium. This syncytium covers the placenta and manages the exchange of nutrients and gases between maternal and fetal circulation. We recently reported protein kinase A (PKA) to be part of a macromolecular signaling complex with ezrin and gap junction protein connexin 43 (Cx43) that provides cAMP-mediated control of gap junction communication. Here, we examined the associated phosphorylation events. Inhibition of PKA activity resulted in decreased Cx43 phosphorylation, which was associated with reduced trophoblast fusion and differentiation. studies using peptide arrays, together with mass spectrometry, pointed to serine 369 and 373 of Cx43 as the major PKA phosphorylation sites that increases gap junction assembly at the plasmalemma. A combination of knockdown and reconstitution experiments and gap-fluorescence loss in photobleaching assays with mutant Cx43 containing single or double phosphoserine-mimicking amino acid substitutions in putative PKA phosphorylation sites demonstrated that phosphorylation of S369 and S373 mediated gap junction communication, trophoblast differentiation, and cell fusion.

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Section: Introductionmentioning

confidence: 99%

Ezrin-anchored PKA phosphorylates serine 369 and 373 on connexin 43 to enhance gap junction assembly, communication, and cell fusion

Dukic

Gerbaud

Gadrey

et al. 2018

Biochemical Journal

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“…This process of syncytialization is a hallmark of placentation and plays a central role in physiological function of the placental barrier as a key regulator of material exchange between the maternal and fetal circulations 23 . While the underlying molecular pathways of syncytialization are not fully understood, studies have shown that activation of adenylate cyclase, which is the regulatory subunit of protein kinase A, by 3′,5′-cyclic monophosphate (cAMP) or forskolin induces BeWo cells and primary villous cytotrophoblast cells to fuse and acquire differentiated phenotypes of the syncytiotrophoblast 24,25 .…”

Section: Resultsmentioning

confidence: 99%

A microphysiological model of the human placental barrier

et al. 2016

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During human pregnancy, the fetal circulation is separated from maternal blood in the placenta by two cell layers – the fetal capillary endothelium and placental trophoblast. This placental barrier plays an essential role in fetal development and health by tightly regulating the exchange of endogenous and exogenous materials between the mother and the fetus. Here we present a microengineered device that provides a novel platform to mimic the structural and functional complexity of this specialized tissue in vitro. Our model is created in a multilayered microfluidic system that enables co-culture of human trophoblast cells and human fetal endothelial cells in a physiologically relevant spatial arrangement to replicate the characteristic architecture of the human placental barrier. We have engineered this co-culture model to induce progressive fusion of trophoblast cells and to form a syncytialized epithelium that resembles the syncytiotrophoblast in vivo. Our system also allows the cultured trophoblasts to form dense microvilli under dynamic flow conditions and to reconstitute expression and physiological localization of membrane transport proteins, such as glucose transporters (GLUTs), critical to the barrier function of the placenta. To provide a proof-of-principle for using this microdevice to recapitulate native function of the placental barrier, we demonstrated physiological transport of glucose across the microengineered maternal-fetal interface. Importantly, the rate of maternal-to-fetal glucose transfer in this system closely approximated that measured in ex vivo perfused human placentas. Our “placenta-on-a-chip” platform represents an important advance in the development of new technologies to model and study the physiological complexity of the human placenta for a wide variety of applications.

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“…The role of P-gp in trophoblast fusion is less surprising as a number of studies have reported the loss or down-regulation of P-gp in the spontaneous differentiation of primary CT to ST. [40][41][42] It is also well known that trophoblast fusion stimulated by the protein kinase A pathway and cAMP involves the induction of GJA1 and hCG. 43 Knockdown of their expression in the BeWo choriocarcinoma model results in an arrest of forskolin-stimulated fusion, 44,45 while overexpression of Cx43 permits Jeg-3 cell fusion. 28 As a point of speculation, beyond its well-known role in substrate distribution and bioavailability and in multidrug resistance, P-gp plays a role in cancer cell invasion, drug resistance and metastasis.…”

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confidence: 99%

P‐Glycoprotein (P‐gp)/ABCB1 plays a functional role in extravillous trophoblast (EVT) invasion and is decreased in the pre‐eclamptic placenta

Dunk

Pappas

Lye

et al. 2018

J Cellular Molecular Medi

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Dysregulation of trophoblast differentiation is implicated in the placental pathologies of intrauterine growth restriction and pre‐eclampsia. P‐glycoprotein (P‐gp encoded by ABCB1) is an ATP‐binding cassette transporter present in the syncytiotrophoblast layer of the placenta where it acts as a molecular sieve. In this study, we show that P‐gp is also expressed in the proliferating cytotrophoblast (CT), the syncytiotrophoblast (ST) and the extravillous trophoblast (EVT), suggesting our hypothesis of a functional role for P‐gp in placental development. Silencing of ABCB1, via siRNA duplex, results in dramatically reduced invasion and migration, and increased tube formation and fusion in the EVT‐like HTR8/SV neo cell line. In both EVT and CT explant differentiation experiments, silencing of ABCB1 leads to induction of the fusion markers human hCG, ERVW‐1 and GJA1 and terminal differentiation of both trophoblast subtypes. Moreover, P‐gp protein levels are decreased in both the villous and the EVT of severe early‐onset pre‐eclamptic placentas. We conclude that, in addition to its role as a syncytial transporter, P‐gp is a key factor in the maintenance of both CT and EVT lineages and that its decrease in severe pre‐eclampsia may contribute to the syncytial and EVT placental pathologies associated with this disease.

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Spatiotemporal regulation of cAMP signaling controls the human trophoblast fusion

Cited by 34 publications

References 82 publications

Ezrin-anchored PKA phosphorylates serine 369 and 373 on connexin 43 to enhance gap junction assembly, communication, and cell fusion

Ezrin-anchored PKA phosphorylates serine 369 and 373 on connexin 43 to enhance gap junction assembly, communication, and cell fusion

A microphysiological model of the human placental barrier

P‐Glycoprotein (P‐gp)/ABCB1 plays a functional role in extravillous trophoblast (EVT) invasion and is decreased in the pre‐eclamptic placenta

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