AM2/IMD secretion from human pulmonary smooth muscle cells and pulmonary fibroblasts is augmented in response to mechanical stretch

Gavin Thomas, Mark Harbinson, Gary McVeigh, Chris Watson, Malcolm Campbell, Alice Cheung, David Bell

Research output: Contribution to journalMeeting abstract

Abstract

Introduction: Pulmonary hypertension (PHT) is a severe life-limiting condition resulting in progressive shortness of breath, exercise intolerance and heart failure. PHT is defined by increased mean pulmonary arterial pressure (PAP) ≥ 25mmHg at rest, and has been attributed to an imbalance between vasodilator and vasoconstrictor influences in the pulmonary microcirculation. Assessment of the vasodilator AM2/IMD, a member of the CGRP/AM peptide family, may have potential application as novel disease biomarker. Objective: to quantify secretion of AM2/IMD from human pulmonary vascular cells cultured under basal, simulated normotensive and hypertensive conditions. Methods: pulmonary fibroblasts (PF), pulmonary smooth muscle (PSM), human pulmonary artery endothelial cells (HPAEC) and human pulmonary microvascular endothelial cells (HPMEC) were cultured on silicone elastomer-bottomed Flexcell plates pre-coated with Matrigel® at rest (un-flexed) or subjected to cyclic mechanical stretch (Flexcell Strain Unit) to simulate pulmonary normotensive (15mmHg, 2.0kPA) and hypertensive (40mmHg, 5.3kPA) conditions at a frequency of 1 Hz (60 cycles per minute) for 48 h. AM2/IMD was extracted from the medium of cultured cells and quantified by ELISA (Phoenix Pharmaceuticals Inc. Karlsruhe, Germany). Results: concentrations of AM2/IMD in culture medium from cells incubated under various conditions were as follows: ng.ml-1, mean + SE, n=2-12; *difference relative to un-flexed, P<0.05; + difference between normotensive and hypertensive condition. PF PSM HPAEC HPMEC un-flexed 6.98+ 2.01 0.40+ 0.06 1.28+0.24 12.63+1.38 normotensive 51.49+11.25* 106.81+59.22 0.48+0.06* 13.53+1.67 hypertensive 41.58+ 8.57* 83.65+20.53* 0.82+0.07+ 18.96+2.43* Conclusion: cyclic stretch enhanced secretion of AM2/IMD from PF and PSM, indicating that these cells may be an important source of this vasodilator peptide in the pulmonary microcirculation under physiological conditions. Secretion was not augmented in hypertension relative to normotensive conditions. AM2/IMD is unlikely therefore to be a suitable diagnostic or prognostic biomarker in PHT.
Original languageEnglish
JournalHeart (British Cardiac Society)
Volume104
Issue numberSuppl 4
DOIs
Publication statusPublished - 26 Mar 2018

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Smooth Muscle Myocytes
Fibroblasts
Lung
Vasodilator Agents
Pulmonary Hypertension
Endothelial Cells
Microcirculation
Pulmonary Artery
Smooth Muscle
Cultured Cells
Biomarkers
Silicone Elastomers
Peptides
Vasoconstrictor Agents
Dyspnea
Germany
Blood Vessels
Culture Media
Arterial Pressure
Heart Failure

Cite this

@article{3e1a1f9c8f9b44f697c5e66766d13be0,
title = "AM2/IMD secretion from human pulmonary smooth muscle cells and pulmonary fibroblasts is augmented in response to mechanical stretch",
abstract = "Introduction: Pulmonary hypertension (PHT) is a severe life-limiting condition resulting in progressive shortness of breath, exercise intolerance and heart failure. PHT is defined by increased mean pulmonary arterial pressure (PAP) ≥ 25mmHg at rest, and has been attributed to an imbalance between vasodilator and vasoconstrictor influences in the pulmonary microcirculation. Assessment of the vasodilator AM2/IMD, a member of the CGRP/AM peptide family, may have potential application as novel disease biomarker. Objective: to quantify secretion of AM2/IMD from human pulmonary vascular cells cultured under basal, simulated normotensive and hypertensive conditions. Methods: pulmonary fibroblasts (PF), pulmonary smooth muscle (PSM), human pulmonary artery endothelial cells (HPAEC) and human pulmonary microvascular endothelial cells (HPMEC) were cultured on silicone elastomer-bottomed Flexcell plates pre-coated with Matrigel{\circledR} at rest (un-flexed) or subjected to cyclic mechanical stretch (Flexcell Strain Unit) to simulate pulmonary normotensive (15mmHg, 2.0kPA) and hypertensive (40mmHg, 5.3kPA) conditions at a frequency of 1 Hz (60 cycles per minute) for 48 h. AM2/IMD was extracted from the medium of cultured cells and quantified by ELISA (Phoenix Pharmaceuticals Inc. Karlsruhe, Germany). Results: concentrations of AM2/IMD in culture medium from cells incubated under various conditions were as follows: ng.ml-1, mean + SE, n=2-12; *difference relative to un-flexed, P<0.05; + difference between normotensive and hypertensive condition. PF PSM HPAEC HPMEC un-flexed 6.98+ 2.01 0.40+ 0.06 1.28+0.24 12.63+1.38 normotensive 51.49+11.25* 106.81+59.22 0.48+0.06* 13.53+1.67 hypertensive 41.58+ 8.57* 83.65+20.53* 0.82+0.07+ 18.96+2.43* Conclusion: cyclic stretch enhanced secretion of AM2/IMD from PF and PSM, indicating that these cells may be an important source of this vasodilator peptide in the pulmonary microcirculation under physiological conditions. Secretion was not augmented in hypertension relative to normotensive conditions. AM2/IMD is unlikely therefore to be a suitable diagnostic or prognostic biomarker in PHT.",
author = "Gavin Thomas and Mark Harbinson and Gary McVeigh and Chris Watson and Malcolm Campbell and Alice Cheung and David Bell",
year = "2018",
month = "3",
day = "26",
doi = "10.1136/heartjnl-2018-SCF.20",
language = "English",
volume = "104",
journal = "Heart (British Cardiac Society)",
issn = "1355-6037",
publisher = "BMJ Publishing Group",
number = "Suppl 4",

}

TY - JOUR

T1 - AM2/IMD secretion from human pulmonary smooth muscle cells and pulmonary fibroblasts is augmented in response to mechanical stretch

AU - Thomas, Gavin

AU - Harbinson, Mark

AU - McVeigh, Gary

AU - Watson, Chris

AU - Campbell, Malcolm

AU - Cheung, Alice

AU - Bell, David

PY - 2018/3/26

Y1 - 2018/3/26

N2 - Introduction: Pulmonary hypertension (PHT) is a severe life-limiting condition resulting in progressive shortness of breath, exercise intolerance and heart failure. PHT is defined by increased mean pulmonary arterial pressure (PAP) ≥ 25mmHg at rest, and has been attributed to an imbalance between vasodilator and vasoconstrictor influences in the pulmonary microcirculation. Assessment of the vasodilator AM2/IMD, a member of the CGRP/AM peptide family, may have potential application as novel disease biomarker. Objective: to quantify secretion of AM2/IMD from human pulmonary vascular cells cultured under basal, simulated normotensive and hypertensive conditions. Methods: pulmonary fibroblasts (PF), pulmonary smooth muscle (PSM), human pulmonary artery endothelial cells (HPAEC) and human pulmonary microvascular endothelial cells (HPMEC) were cultured on silicone elastomer-bottomed Flexcell plates pre-coated with Matrigel® at rest (un-flexed) or subjected to cyclic mechanical stretch (Flexcell Strain Unit) to simulate pulmonary normotensive (15mmHg, 2.0kPA) and hypertensive (40mmHg, 5.3kPA) conditions at a frequency of 1 Hz (60 cycles per minute) for 48 h. AM2/IMD was extracted from the medium of cultured cells and quantified by ELISA (Phoenix Pharmaceuticals Inc. Karlsruhe, Germany). Results: concentrations of AM2/IMD in culture medium from cells incubated under various conditions were as follows: ng.ml-1, mean + SE, n=2-12; *difference relative to un-flexed, P<0.05; + difference between normotensive and hypertensive condition. PF PSM HPAEC HPMEC un-flexed 6.98+ 2.01 0.40+ 0.06 1.28+0.24 12.63+1.38 normotensive 51.49+11.25* 106.81+59.22 0.48+0.06* 13.53+1.67 hypertensive 41.58+ 8.57* 83.65+20.53* 0.82+0.07+ 18.96+2.43* Conclusion: cyclic stretch enhanced secretion of AM2/IMD from PF and PSM, indicating that these cells may be an important source of this vasodilator peptide in the pulmonary microcirculation under physiological conditions. Secretion was not augmented in hypertension relative to normotensive conditions. AM2/IMD is unlikely therefore to be a suitable diagnostic or prognostic biomarker in PHT.

AB - Introduction: Pulmonary hypertension (PHT) is a severe life-limiting condition resulting in progressive shortness of breath, exercise intolerance and heart failure. PHT is defined by increased mean pulmonary arterial pressure (PAP) ≥ 25mmHg at rest, and has been attributed to an imbalance between vasodilator and vasoconstrictor influences in the pulmonary microcirculation. Assessment of the vasodilator AM2/IMD, a member of the CGRP/AM peptide family, may have potential application as novel disease biomarker. Objective: to quantify secretion of AM2/IMD from human pulmonary vascular cells cultured under basal, simulated normotensive and hypertensive conditions. Methods: pulmonary fibroblasts (PF), pulmonary smooth muscle (PSM), human pulmonary artery endothelial cells (HPAEC) and human pulmonary microvascular endothelial cells (HPMEC) were cultured on silicone elastomer-bottomed Flexcell plates pre-coated with Matrigel® at rest (un-flexed) or subjected to cyclic mechanical stretch (Flexcell Strain Unit) to simulate pulmonary normotensive (15mmHg, 2.0kPA) and hypertensive (40mmHg, 5.3kPA) conditions at a frequency of 1 Hz (60 cycles per minute) for 48 h. AM2/IMD was extracted from the medium of cultured cells and quantified by ELISA (Phoenix Pharmaceuticals Inc. Karlsruhe, Germany). Results: concentrations of AM2/IMD in culture medium from cells incubated under various conditions were as follows: ng.ml-1, mean + SE, n=2-12; *difference relative to un-flexed, P<0.05; + difference between normotensive and hypertensive condition. PF PSM HPAEC HPMEC un-flexed 6.98+ 2.01 0.40+ 0.06 1.28+0.24 12.63+1.38 normotensive 51.49+11.25* 106.81+59.22 0.48+0.06* 13.53+1.67 hypertensive 41.58+ 8.57* 83.65+20.53* 0.82+0.07+ 18.96+2.43* Conclusion: cyclic stretch enhanced secretion of AM2/IMD from PF and PSM, indicating that these cells may be an important source of this vasodilator peptide in the pulmonary microcirculation under physiological conditions. Secretion was not augmented in hypertension relative to normotensive conditions. AM2/IMD is unlikely therefore to be a suitable diagnostic or prognostic biomarker in PHT.

U2 - 10.1136/heartjnl-2018-SCF.20

DO - 10.1136/heartjnl-2018-SCF.20

M3 - Meeting abstract

VL - 104

JO - Heart (British Cardiac Society)

JF - Heart (British Cardiac Society)

SN - 1355-6037

IS - Suppl 4

ER -