Transcriptional profiling of the ductus arteriosus: Comparison of rodent microarrays and human RNA sequencing

Yarboro, Michael T.; Durbin, Matthew D.; Herington, Jennifer L.; Shelton, Elaine L.; Zhang, Tao; Ebby, Cris G; Stoller, Jason Z.; Clyman, Ronald I.; Reese, Jeff

doi:10.1053/j.semperi.2018.05.003

Cited by 15 publications

(27 citation statements)

References 78 publications

(99 reference statements)

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“…We evaluated the activity of VU0071063 on Kir6.2/SUR1 and Kir6.1/SUR2B channels heterologously expressed in HEK-293 cells, as well as on native Kir6.1/ SUR2B expressed in ductus arteriosus vessels. They were chosen for these studies because they are enriched in vascular K ATP channels composed of Kir6.1 and SUR2B (Shelton et al, 2014;Yarboro et al, 2018). As shown in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Pressure Myography. Ductus arteriosus (DA) vessels, which are enriched in Kir6.1/SUR2B channels (Shelton et al, 2014;Yarboro et al, 2018), were isolated from term-gestation fetal mice and mounted on glass pipette tips submerged in microvessel perfusion chambers as previously reported (Reese et al, 2009;Pfaltzgraff et al, 2014;Hooper et al, 2016). Chambers were placed on inverted microscopes equipped with IonOptix digital image capture and dimensioning software.…”

Section: Methodsmentioning

confidence: 99%

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Structure-Activity Relationships, Pharmacokinetics, and Pharmacodynamics of the Kir6.2/SUR1-Specific Channel Opener VU0071063

Kharade¹,

Sánchez-Andrés²,

Fulton

et al. 2019

J Pharmacol Exp Ther

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Glucose-stimulated insulin secretion from pancreatic b-cells is controlled by ATP-regulated potassium (K ATP) channels composed of Kir6.2 and sulfonylurea receptor 1 (SUR1) subunits. The K ATP channel-opener diazoxide is FDA-approved for treating hyperinsulinism and hypoglycemia but suffers from off-target effects on vascular K ATP channels and other ion channels. The development of more specific openers would provide critically needed tool compounds for probing the therapeutic potential of Kir6.2/SUR1 activation. Here, we characterize a novel scaffold activator of Kir6.2/SUR1 that our group recently discovered in a high-throughput screen. Optimization efforts with medicinal chemistry identified key structural elements that are essential for VU0071063-dependent opening of Kir6.2/SUR1. VU0071063 has no effects on heterologously expressed Kir6.1/SUR2B channels or ductus arteriole tone, indicating it does not open vascular K ATP channels. VU0071063 induces hyperpolarization of b-cell membrane potential and inhibits insulin secretion more potently than diazoxide. VU0071063 exhibits metabolic and pharmacokinetic properties that are favorable for an in vivo probe and is brain penetrant. Administration of VU0071063 inhibits glucose-stimulated insulin secretion and glucose-lowering in mice. Taken together, these studies indicate that VU0071063 is a more potent and specific opener of Kir6.2/SUR1 than diazoxide and should be useful as an in vitro and in vivo tool compound for investigating the therapeutic potential of Kir6.2/SUR1 expressed in the pancreas and brain.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Structure-Activity Relationships, Pharmacokinetics, and Pharmacodynamics of the Kir6.2/SUR1-Specific Channel Opener VU0071063

Kharade¹,

Sánchez-Andrés²,

Fulton

et al. 2019

J Pharmacol Exp Ther

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“…A systematic review of transcriptional profiling in the rodent and human DA in comparison to the aorta demonstrated that a higher expression of EP4 was observed commonly in the rodent and human DA. 41 The differential EP4 expression was observed in mice and humans at embryonic day 14 and 21 weeks of gestation, respectively, 25 , 41 and EP4 expression increases toward term. 9 , 25 , 42 , 43 The present study identified fibulin-1 as the gene regulated by EP4 signaling, and fibulin-1 expression may be upregulated in accordance with that of EP4 expression.…”

Section: Discussionmentioning

confidence: 99%

Fibulin-1 Integrates Subendothelial Extracellular Matrices and Contributes to Anatomical Closure of the Ductus Arteriosus

Ito

Yokoyama

Nakakoji

et al. 2020

ATVB

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Objective: The ductus arteriosus (DA) is a fetal artery connecting the aorta and pulmonary arteries. Progressive matrix remodeling, that is, intimal thickening (IT), occurs in the subendothelial region of DA to bring anatomic DA closure. IT is comprised of multiple ECMs (extracellular matrices) and migrated smooth muscle cells (SMCs). Because glycoprotein fibulin-1 binds to multiple ECMs and regulates morphogenesis during development, we investigated the role of fibulin-1 in DA closure. Approach and Results: Fibulin-1–deficient ( Fbln1 −/− ) mice exhibited patent DA with hypoplastic IT. An unbiased transcriptome analysis revealed that EP4 (prostaglandin E receptor 4) stimulation markedly increased fibulin-1 in DA-SMCs via phospholipase C-NFκB (nuclear factor κB) signaling pathways. Fluorescence-activated cell sorting (FACS) analysis demonstrated that fibulin-1 binding protein versican was derived from DA-endothelial cells (ECs). We examined the effect of fibulin-1 on directional migration toward ECs in association with versican by using cocultured DA-SMCs and ECs. EP4 stimulation promoted directional DA-SMC migration toward ECs, which was attenuated by either silencing fibulin-1 or versican. Immunofluorescence demonstrated that fibulin-1 and versican V0/V1 were coexpressed at the IT of wild-type DA, whereas 30% of versican-deleted mice lacking a hyaluronan binding site displayed patent DA. Fibulin-1 expression was attenuated in the EP4-deficient mouse ( Ptger4 −/− ) DA, which exhibits patent DA with hypoplastic IT, and fibulin-1 protein administration restored IT formation. In human DA, fibulin-1 and versican were abundantly expressed in SMCs and ECs, respectively. Conclusions: Fibulin-1 contributes to DA closure by forming an environment favoring directional SMC migration toward the subendothelial region, at least, in part, in combination with EC-derived versican and its binding partner hyaluronan.

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“…43 We further examined the expression of t-PA in human DA. 43 Although RNA sequencing of the human DA was recently reported, 46 there has been no transcriptional profiling of the human DA intimal thickening region that contains DA endothelial cells. We collected human DA from neonatal patients with congenital heart disease and examined the expression of t-PA in the intimal thickening region compared with the tunica media.…”

Section: Expression Of T-pa In Da Endothelial Cellsmentioning

confidence: 99%

Role of Tissue-Type Plasminogen Activator in Remodeling of the Ductus Arteriosus

2020

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pO2, decreased blood pressure in the DA lumen, decreased circulating prostaglandin E2 (PGE2) level, and a decreased number of PGE2 receptors. 6-8The postnatal increase of pO2 inhibits the ductal smooth muscle voltage-dependent potassium channels, such as Kv1.5 and Kv2.1, 9,10 which causes membrane depolarization and calcium influx via the voltage-dependent calcium channels. This is followed by DA SMC contraction. 11 Voltage-dependent calcium channel and potassium channel genes are upregulated in the mature DA. 12,13 In addition to calcium influx via voltage-dependent calcium channels, oxygen-induced activation of the Rho/Rho-associated coiled-coil containing protein kinase (Rho/ROCK) pathway is associated with postnatal DA constriction. 14 Reduced circulating PGE2 is related to postnatal DA constriction. The fetal DA is dilated by PGE2, which is derived from the placenta. 15 After birth, the loss of the placenta and the increased flow of the lung, which is the major site of PGE catabolism, cause a decline in circulating PGE2. 16 In addition, PGE2 receptors are decreased after birth. 17 These changes promote DA contraction.Furthermore, we recently reported that serum osmotic change and glutamate are related to postnatal DA constriction. 18, 19 We found that premature infants did not have a physiological decrease in serum osmolarity after birth, and Vascular remodeling (e.g., intimal thickening) is necessary for complete closure of the ductus arteriosus (DA). Smooth muscle cells are reported to contribute to DA remodeling. In contrast, the contribution of endothelial cells remains largely unknown. Recent data showed that tissue-type plasminogen activator (t-PA) was highly expressed in the endothelial cells of rat and human DA. It is well known that t-PA is an activator of the blood fibrinolytic system, but t-PA-induced localized proteolysis has been reported to play an important role in vascular development. We found that t-PA-induced plasminogen-plasmin conversion promoted matrix metalloproteinase-2 activation in endothelial cells of rat DA. Gelatinase activity was noted at the internal elastic laminae (IEL) of rat and human DA on in situ gelatin zymography. The in vivo injection of plasminogen to pre-term rats increased gelatinase activation, IEL disruption, and the subsequent intimal thickening formation in the pre-term rat DA. Human DA results partly supported the rat DA findings, suggesting that t-PA-mediated DA remodeling may also be present in the human DA. Current pharmacotherapy for patent DA (PDA) mainly focuses on increasing vascular constriction. Elucidating the molecular mechanisms of DA remodeling may help to expand the range of therapeutic strategies for PDA.

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Transcriptional profiling of the ductus arteriosus: Comparison of rodent microarrays and human RNA sequencing

Cited by 15 publications

References 78 publications

Structure-Activity Relationships, Pharmacokinetics, and Pharmacodynamics of the Kir6.2/SUR1-Specific Channel Opener VU0071063

Structure-Activity Relationships, Pharmacokinetics, and Pharmacodynamics of the Kir6.2/SUR1-Specific Channel Opener VU0071063

Fibulin-1 Integrates Subendothelial Extracellular Matrices and Contributes to Anatomical Closure of the Ductus Arteriosus

Role of Tissue-Type Plasminogen Activator in Remodeling of the Ductus Arteriosus

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