RNA interference targeted to multiple P2X receptor subtypes attenuates zinc-induced calcium entry

Lihua Liang, A. Zsembery, Erik M. Schwiebert

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

A postulated therapeutic avenue in cystic fibrosis (CF) is activation of Ca2+-dependent Cl- channels via stimulation of Ca 2+ entry from extracellular solutions independent of CFTR functional status. We have shown that extracellular zinc and ATP induce a sustained increase in cytosolic Ca2+ in human airway epithelial cells that translates into stimulation of sustained secretory Cl- transport in non-CF and CF human and mouse airway epithelial cells, cell monolayers, and nasal mucosa. On the basis of these studies, the Ca2+ entry channels most likely involved were P2X purinergic receptor channels. In the present study, molecular and biochemical data show coexpression of P2X4, P2X5, and P2X6 subtypes in non-CF (16HBE14 o-) and CF (IB3-1) human bronchial epithelial cells. Other P2X receptor Ca2+ entry channel subtypes are expressed rarely or not at all in airway epithelia, epithelial cell models from other CF-relevant tissues, or vascular endothelia. Novel transient lipid transfection-mediated delivery of small interference RNA fragments specific to P2X4 and P2X6 (but not P2X5) into IB3-1 CF human airway epithelial cells inhibited extracellular zinc- and ATP-induced Ca2+ entry markedly in fura-2 Ca2+ measurements and "knocked down" protein by >65%. These data suggest that multiple P2X receptor Ca 2+ entry channel subtypes are expressed in airway epithelia. P2X 4 and P2X6 may coassemble on the airway surface as targets for possible therapeutics for CF independent of CFTR genotype.

Original languageEnglish
JournalAmerican Journal of Physiology - Cell Physiology
Volume289
Issue number2 58-2
DOIs
Publication statusPublished - Aug 2005

Fingerprint

RNA Interference
Cystic Fibrosis
Zinc
RNA
Calcium
Epithelial Cells
Purinergic P2X Receptors
Fibrosis
Epithelium
Adenosine Triphosphate
Nasal Mucosa
Fura-2
Vascular Endothelium
Transfection
Monolayers
Chemical activation
Genotype
Tissue
Lipids
Therapeutics

Keywords

  • Airway epithelia
  • Cystic fibrosis
  • Purinergic receptors
  • Therapy
  • Zinc receptors

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

RNA interference targeted to multiple P2X receptor subtypes attenuates zinc-induced calcium entry. / Liang, Lihua; Zsembery, A.; Schwiebert, Erik M.

In: American Journal of Physiology - Cell Physiology, Vol. 289, No. 2 58-2, 08.2005.

Research output: Contribution to journalArticle

@article{b430da9cf88d4b2d9e3bd5d1d7748fb4,
title = "RNA interference targeted to multiple P2X receptor subtypes attenuates zinc-induced calcium entry",
abstract = "A postulated therapeutic avenue in cystic fibrosis (CF) is activation of Ca2+-dependent Cl- channels via stimulation of Ca 2+ entry from extracellular solutions independent of CFTR functional status. We have shown that extracellular zinc and ATP induce a sustained increase in cytosolic Ca2+ in human airway epithelial cells that translates into stimulation of sustained secretory Cl- transport in non-CF and CF human and mouse airway epithelial cells, cell monolayers, and nasal mucosa. On the basis of these studies, the Ca2+ entry channels most likely involved were P2X purinergic receptor channels. In the present study, molecular and biochemical data show coexpression of P2X4, P2X5, and P2X6 subtypes in non-CF (16HBE14 o-) and CF (IB3-1) human bronchial epithelial cells. Other P2X receptor Ca2+ entry channel subtypes are expressed rarely or not at all in airway epithelia, epithelial cell models from other CF-relevant tissues, or vascular endothelia. Novel transient lipid transfection-mediated delivery of small interference RNA fragments specific to P2X4 and P2X6 (but not P2X5) into IB3-1 CF human airway epithelial cells inhibited extracellular zinc- and ATP-induced Ca2+ entry markedly in fura-2 Ca2+ measurements and {"}knocked down{"} protein by >65{\%}. These data suggest that multiple P2X receptor Ca 2+ entry channel subtypes are expressed in airway epithelia. P2X 4 and P2X6 may coassemble on the airway surface as targets for possible therapeutics for CF independent of CFTR genotype.",
keywords = "Airway epithelia, Cystic fibrosis, Purinergic receptors, Therapy, Zinc receptors",
author = "Lihua Liang and A. Zsembery and Schwiebert, {Erik M.}",
year = "2005",
month = "8",
doi = "10.1152/ajpcell.00491.2004",
language = "English",
volume = "289",
journal = "American Journal of Physiology",
issn = "0363-6119",
publisher = "American Physiological Society",
number = "2 58-2",

}

TY - JOUR

T1 - RNA interference targeted to multiple P2X receptor subtypes attenuates zinc-induced calcium entry

AU - Liang, Lihua

AU - Zsembery, A.

AU - Schwiebert, Erik M.

PY - 2005/8

Y1 - 2005/8

N2 - A postulated therapeutic avenue in cystic fibrosis (CF) is activation of Ca2+-dependent Cl- channels via stimulation of Ca 2+ entry from extracellular solutions independent of CFTR functional status. We have shown that extracellular zinc and ATP induce a sustained increase in cytosolic Ca2+ in human airway epithelial cells that translates into stimulation of sustained secretory Cl- transport in non-CF and CF human and mouse airway epithelial cells, cell monolayers, and nasal mucosa. On the basis of these studies, the Ca2+ entry channels most likely involved were P2X purinergic receptor channels. In the present study, molecular and biochemical data show coexpression of P2X4, P2X5, and P2X6 subtypes in non-CF (16HBE14 o-) and CF (IB3-1) human bronchial epithelial cells. Other P2X receptor Ca2+ entry channel subtypes are expressed rarely or not at all in airway epithelia, epithelial cell models from other CF-relevant tissues, or vascular endothelia. Novel transient lipid transfection-mediated delivery of small interference RNA fragments specific to P2X4 and P2X6 (but not P2X5) into IB3-1 CF human airway epithelial cells inhibited extracellular zinc- and ATP-induced Ca2+ entry markedly in fura-2 Ca2+ measurements and "knocked down" protein by >65%. These data suggest that multiple P2X receptor Ca 2+ entry channel subtypes are expressed in airway epithelia. P2X 4 and P2X6 may coassemble on the airway surface as targets for possible therapeutics for CF independent of CFTR genotype.

AB - A postulated therapeutic avenue in cystic fibrosis (CF) is activation of Ca2+-dependent Cl- channels via stimulation of Ca 2+ entry from extracellular solutions independent of CFTR functional status. We have shown that extracellular zinc and ATP induce a sustained increase in cytosolic Ca2+ in human airway epithelial cells that translates into stimulation of sustained secretory Cl- transport in non-CF and CF human and mouse airway epithelial cells, cell monolayers, and nasal mucosa. On the basis of these studies, the Ca2+ entry channels most likely involved were P2X purinergic receptor channels. In the present study, molecular and biochemical data show coexpression of P2X4, P2X5, and P2X6 subtypes in non-CF (16HBE14 o-) and CF (IB3-1) human bronchial epithelial cells. Other P2X receptor Ca2+ entry channel subtypes are expressed rarely or not at all in airway epithelia, epithelial cell models from other CF-relevant tissues, or vascular endothelia. Novel transient lipid transfection-mediated delivery of small interference RNA fragments specific to P2X4 and P2X6 (but not P2X5) into IB3-1 CF human airway epithelial cells inhibited extracellular zinc- and ATP-induced Ca2+ entry markedly in fura-2 Ca2+ measurements and "knocked down" protein by >65%. These data suggest that multiple P2X receptor Ca 2+ entry channel subtypes are expressed in airway epithelia. P2X 4 and P2X6 may coassemble on the airway surface as targets for possible therapeutics for CF independent of CFTR genotype.

KW - Airway epithelia

KW - Cystic fibrosis

KW - Purinergic receptors

KW - Therapy

KW - Zinc receptors

UR - http://www.scopus.com/inward/record.url?scp=22444433901&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=22444433901&partnerID=8YFLogxK

U2 - 10.1152/ajpcell.00491.2004

DO - 10.1152/ajpcell.00491.2004

M3 - Article

VL - 289

JO - American Journal of Physiology

JF - American Journal of Physiology

SN - 0363-6119

IS - 2 58-2

ER -