Targeting Receptor Tyrosine Kinases and Their Downstream Signaling with Cell-Penetrating Peptides in Human Pulmonary Artery Smooth Muscle and Endothelial Cells

Thrombin is a procoagulant inflammatory agonist that can disrupt the endothelium-lumen barrier in the lung by causing contraction of endothelial cells and promote pulmonary cell proliferation. Both contraction and proliferation require increases in cytosolic Ca2+ concentration ([Ca2+]cyt). In this study, we compared the effect of thrombin on Ca2+ signaling in human pulmonary artery smooth muscle (PASMC) and endothelial (PAEC) cells. Thrombin increased the [Ca2+]cyt in both cell types; however, the transient response was significantly higher and recovered quicker in the PASMC, suggesting different mechanisms may contribute to thrombin-mediated increases in [Ca2+]cyt in these cell types. Depletion of intracellular stores with cyclopiazonic acid (CPA) in the absence of extracellular Ca2+ induced calcium transients representative of those observed in response to thrombin in both cell types. Interestingly, CPA pretreatment significantly attenuated thrombin-induced Ca2+ release in PASMC; this attenuation was not apparent in PAEC, indicating that a PAEC-specific mechanism was targeted by thrombin. Treatment with a combination of CPA, caffeine, and ryanodine also failed to abolish the thrombin-induced Ca2+ transient in PAEC. Notably, thrombin-induced receptor-mediated calcium influx was still observed in PASMC after CPA pretreatment in the presence of extracellular Ca2+. Ca2+ oscillations were triggered by thrombin in PASMC resulting from a balance of extracellular Ca2+ influx and Ca2+ reuptake by the sarcoplasmic reticulum. The data show that thrombin induces increases in intracellular calcium in PASMC and PAEC with a distinct CPA-, caffeine-, and ryanodine-insensitive release existing only in PAEC. Furthermore, a dynamic balance between Ca2+ influx, intracellular Ca2+ release, and reuptake underlie the Ca2+ transients evoked by thrombin in some PASMC. Understanding of such mechanisms will provide an important insight into thrombin-mediated vascular injury during hypertension.

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S112 Conditioned media from human pulmonary arterial endothelial cells treated with hepcidin or haemoglobin cause proliferation and migration of human pulmonary artery smooth muscle cells

10.1136/thoraxjnl-2017-210983.118 ◽

2017 ◽

Author(s):

T Shackshaft ◽

SJ Wort ◽

GJ Quinlan ◽

L Ramakrishnan

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Pulmonary Artery ◽

Smooth Muscle Cells ◽

Pulmonary Arterial ◽

Pulmonary Artery Smooth Muscle ◽

Human Pulmonary Artery ◽

And Migration ◽

Arterial Endothelial Cells ◽

Proliferation And Migration

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DDAH1 Overexpression in Human Pulmonary Vascular Endothelial Cells Co‐Cultured with Human Pulmonary Artery Smooth Muscle Cells

The FASEB Journal ◽

10.1096/fasebj.2020.34.s1.01937 ◽

2020 ◽

Vol 34 (S1) ◽

pp. 1-1

Author(s):

Jennifer K. Trittmann ◽

Hanadi Almazroue ◽

Yi Jin ◽

John Barba ◽

Avante D. Milton ◽

...

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Pulmonary Artery ◽

Smooth Muscle Cells ◽

Vascular Endothelial Cells ◽

Muscle Cells ◽

Vascular Endothelial ◽

Pulmonary Artery Smooth Muscle ◽

Human Pulmonary Artery

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Extracellular matrix degradation pathways and fatty acid metabolism regulate distinct pulmonary vascular cell types in pulmonary arterial hypertension

Pulmonary Circulation ◽

10.1177/2045894021996190 ◽

2021 ◽

Vol 11 (1) ◽

pp. 204589402199619

Author(s):

Sharon Mumby ◽

F. Perros ◽

C. Hui ◽

B.L. Xu ◽

W. Xu ◽

...

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Pulmonary Arterial Hypertension ◽

Pulmonary Artery ◽

Arterial Hypertension ◽

Cell Types ◽

Microvascular Endothelial Cells ◽

Pulmonary Arterial ◽

Pulmonary Artery Smooth Muscle ◽

Human Pulmonary Artery

Pulmonary arterial hypertension describes a group of diseases characterised by raised pulmonary vascular resistance, resulting from vascular remodelling in the pre-capillary resistance arterioles. Left untreated, patients die from right heart failure. Pulmonary vascular remodelling involves all cell types but to date the precise roles of the different cells is unknown. This study investigated differences in basal gene expression between pulmonary arterial hypertension and controls using both human pulmonary microvascular endothelial cells and human pulmonary artery smooth muscle cells. Human pulmonary microvascular endothelial cells and human pulmonary artery smooth muscle cells from pulmonary arterial hypertension patients and controls were cultured to confluence, harvested and RNA extracted. Whole genome sequencing was performed and after transcript quantification and normalisation, we examined differentially expressed genes and applied gene set enrichment analysis to the differentially expressed genes to identify putative activated pathways. Human pulmonary microvascular endothelial cells displayed 1008 significant ( p ≤ 0.0001) differentially expressed genes in pulmonary arterial hypertension samples compared to controls. In human pulmonary artery smooth muscle cells, there were 229 significant ( p ≤ 0.0001) differentially expressed genes between pulmonary arterial hypertension and controls. Pathway analysis revealed distinctive differences: human pulmonary microvascular endothelial cells display down-regulation of extracellular matrix organisation, collagen formation and biosynthesis, focal- and cell-adhesion molecules suggesting severe endothelial barrier dysfunction and vascular permeability in pulmonary arterial hypertension pathogenesis. In contrast, pathways in human pulmonary artery smooth muscle cells were mainly up-regulated, including those for fatty acid metabolism, biosynthesis of unsaturated fatty acids, cell–cell and adherens junction interactions suggesting a more energy-driven proliferative phenotype. This suggests that the two cell types play different mechanistic roles in pulmonary arterial hypertension pathogenesis and further studies are required to fully elucidate the role each plays and the interactions between these cell types in vascular remodelling in disease progression.

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S55 Effect of electronic cigarette extraction on vasoactive gene expression in human pulmonary artery smooth muscle and endothelial cells

10.1136/thorax-2018-212555.61 ◽

2018 ◽

Author(s):

A Alqarni ◽

O Brand ◽

P Alice ◽

M Alshehri ◽

L Pang

Keyword(s):

Gene Expression ◽

Endothelial Cells ◽

Smooth Muscle ◽

Pulmonary Artery ◽

Electronic Cigarette ◽

Pulmonary Artery Smooth Muscle ◽

Human Pulmonary Artery

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Histamine-mediated increases in cytosolic [Ca2+] involve different mechanisms in human pulmonary artery smooth muscle and endothelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00236.2005 ◽

2006 ◽

Vol 290 (2) ◽

pp. C325-C336 ◽

Cited By ~ 11

Author(s):

Joseph R. H. Mauban ◽

Katherine Wilkinson ◽

Christian Schach ◽

Jason X.-J. Yuan

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Pulmonary Artery ◽

Solution Treatment ◽

Cyclopiazonic Acid ◽

Free Solution ◽

Intracellular Ca ◽

Pulmonary Artery Smooth Muscle ◽

Human Pulmonary Artery ◽

Stimulation Of

Agonist stimulation of human pulmonary artery smooth muscle cells (PASMC) and endothelial cells (PAEC) with histamine showed similar spatiotemporal patterns of Ca2+ release. Both sustained elevation and oscillatory patterns of changes in cytosolic Ca2+ concentration ([Ca2+]cyt) were observed in the absence of extracellular Ca2+. Capacitative Ca2+ entry (CCE) was induced in PASMC and PAEC by passive depletion of intracellular Ca2+ stores with 10 μM cyclopiazonic acid (CPA; 15–30 min). The pyrazole derivative BTP2 inhibited CPA-activated Ca2+ influx, suggesting that depletion of CPA-sensitive internal stores is sufficient to induce CCE in both PASMC and PAEC. The recourse of histamine-mediated Ca2+ release was examined after exposure of cells to CPA, thapsigargin, caffeine, ryanodine, FCCP, or bafilomycin. In PASMC bathed in Ca2+-free solution, treatment with CPA almost abolished histamine-induced rises in [Ca2+]cyt. In PAEC bathed in Ca2+-free solution, however, treatment with CPA eliminated histamine-induced sustained and oscillatory rises in [Ca2+]cyt but did not affect initial transient increase in [Ca2+]cyt. Furthermore, treatment of PAEC with a combination of CPA (or thapsigargin) and caffeine (and ryanodine), FCCP, or bafilomycin did not abolish histamine-induced transient [Ca2+]cyt increases. These observations indicate that 1) depletion of CPA-sensitive stores is sufficient to cause CCE in both PASMC and PAEC; 2) induction of CCE in PAEC does not require depletion of all internal Ca2+ stores; 3) the histamine-releasable internal stores in PASMC are mainly CPA-sensitive stores; 4) PAEC, in addition to a CPA-sensitive functional pool, contain other stores insensitive to CPA, thapsigargin, caffeine, ryanodine, FCCP, and bafilomycin; and 5) although the CPA-insensitive stores in PAEC may not contribute to CCE, they contribute to histamine-mediated Ca2+ release.

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