Evidence against the rescue of defective ΔF508-CFTR cellular processing by curcumin in cell culture and mouse models

Yuanlin Seng, N. D. Sonawane, Danieli Salinas, Liman Qian, Nicoletta Pedemonte, Luis J V Galietta, A. S. Verkman

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

Curcumin, the yellow colored component of the spice turmeric, has been reported to rescue defective ΔF508-cystic fibrosis transmembrane conductance regulator (CFTR) cellular processing in homozygous mutant mice, restoring nasal potential differences and improving survival (Egan, M. E., Pearson, M., Weiner, S. A., Rajendran, V., Rubin, D., Glockner-Pagel, J., Canny, S., Du, K., Lukacs, G. L., and Caplan, M. J. (2004) Science 304, 600-602). Because of the implied potential use of curcumin or similar compounds in the therapy of cystic fibrosis caused by the ΔF508 mutation, we tried to reproduce and extend the pre-clinical data of Egan et al. Fluorometric measurements of iodide influx in Fischer rat thyroid cells expressing ΔF508-CFTR showed no effect of curcumin (1-40 μM) when added for up to 24 h prior to assay in cells grown at 37 °C. Controls, including 27 °C rescue and 4 mM phenylbutyrate at 37 °C, were strongly positive. Also, curcumin did not increase short circuit current in primary cultures of a human airway epithelium homozygous for ΔF508-CFTR with a 27 °C rescue-positive control. Nasal potential differences in mice were measured in response to topical perfusion with serial solutions containing amiloride, low Cl-, and forskolin. Robust low Cl- and forskolin-induced hyperpolarization of 22 ± 3 mV was found in wild type mice, with 2.1 ± 0.4 mV hyperpolarization in ΔF508 homozygous mutant mice. No significant increase in Cl-/forskolin hyperpolarization was seen in any of the 22 ΔF508 mice studied using different curcumin preparations and administration regimens, including that used by Egan et al. Assay of serum curcumin by ethyl acetate extraction followed by liquid chromatography/mass spectrometry indicated a maximum serum concentration of 60 nM, well below that of 5-15 μM, where cellular effects by sarcoplasmic/endoplasmic reticulum calcium pump inhibition are proposed to occur. Our results do not support further evaluation of curcumin for cystic fibrosis therapy.

Original languageEnglish
Pages (from-to)40629-40633
Number of pages5
JournalJournal of Biological Chemistry
Volume279
Issue number39
DOIs
Publication statusPublished - Sep 24 2004

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Curcumin
Cell culture
Cell Culture Techniques
Processing
Colforsin
Nose
Cystic Fibrosis
Assays
Phenylbutyrates
Cells
Curcuma
Spices
Amiloride
Liquid chromatography
Inbred F344 Rats
Sarcoplasmic Reticulum
Iodides
Serum
Liquid Chromatography

ASJC Scopus subject areas

  • Biochemistry

Cite this

Evidence against the rescue of defective ΔF508-CFTR cellular processing by curcumin in cell culture and mouse models. / Seng, Yuanlin; Sonawane, N. D.; Salinas, Danieli; Qian, Liman; Pedemonte, Nicoletta; Galietta, Luis J V; Verkman, A. S.

In: Journal of Biological Chemistry, Vol. 279, No. 39, 24.09.2004, p. 40629-40633.

Research output: Contribution to journalArticle

Seng, Yuanlin ; Sonawane, N. D. ; Salinas, Danieli ; Qian, Liman ; Pedemonte, Nicoletta ; Galietta, Luis J V ; Verkman, A. S. / Evidence against the rescue of defective ΔF508-CFTR cellular processing by curcumin in cell culture and mouse models. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 39. pp. 40629-40633.
@article{0000263acb314c1781ed7ee2009f17bb,
title = "Evidence against the rescue of defective ΔF508-CFTR cellular processing by curcumin in cell culture and mouse models",
abstract = "Curcumin, the yellow colored component of the spice turmeric, has been reported to rescue defective ΔF508-cystic fibrosis transmembrane conductance regulator (CFTR) cellular processing in homozygous mutant mice, restoring nasal potential differences and improving survival (Egan, M. E., Pearson, M., Weiner, S. A., Rajendran, V., Rubin, D., Glockner-Pagel, J., Canny, S., Du, K., Lukacs, G. L., and Caplan, M. J. (2004) Science 304, 600-602). Because of the implied potential use of curcumin or similar compounds in the therapy of cystic fibrosis caused by the ΔF508 mutation, we tried to reproduce and extend the pre-clinical data of Egan et al. Fluorometric measurements of iodide influx in Fischer rat thyroid cells expressing ΔF508-CFTR showed no effect of curcumin (1-40 μM) when added for up to 24 h prior to assay in cells grown at 37 °C. Controls, including 27 °C rescue and 4 mM phenylbutyrate at 37 °C, were strongly positive. Also, curcumin did not increase short circuit current in primary cultures of a human airway epithelium homozygous for ΔF508-CFTR with a 27 °C rescue-positive control. Nasal potential differences in mice were measured in response to topical perfusion with serial solutions containing amiloride, low Cl-, and forskolin. Robust low Cl- and forskolin-induced hyperpolarization of 22 ± 3 mV was found in wild type mice, with 2.1 ± 0.4 mV hyperpolarization in ΔF508 homozygous mutant mice. No significant increase in Cl-/forskolin hyperpolarization was seen in any of the 22 ΔF508 mice studied using different curcumin preparations and administration regimens, including that used by Egan et al. Assay of serum curcumin by ethyl acetate extraction followed by liquid chromatography/mass spectrometry indicated a maximum serum concentration of 60 nM, well below that of 5-15 μM, where cellular effects by sarcoplasmic/endoplasmic reticulum calcium pump inhibition are proposed to occur. Our results do not support further evaluation of curcumin for cystic fibrosis therapy.",
author = "Yuanlin Seng and Sonawane, {N. D.} and Danieli Salinas and Liman Qian and Nicoletta Pedemonte and Galietta, {Luis J V} and Verkman, {A. S.}",
year = "2004",
month = "9",
day = "24",
doi = "10.1074/jbc.M407308200",
language = "English",
volume = "279",
pages = "40629--40633",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "39",

}

TY - JOUR

T1 - Evidence against the rescue of defective ΔF508-CFTR cellular processing by curcumin in cell culture and mouse models

AU - Seng, Yuanlin

AU - Sonawane, N. D.

AU - Salinas, Danieli

AU - Qian, Liman

AU - Pedemonte, Nicoletta

AU - Galietta, Luis J V

AU - Verkman, A. S.

PY - 2004/9/24

Y1 - 2004/9/24

N2 - Curcumin, the yellow colored component of the spice turmeric, has been reported to rescue defective ΔF508-cystic fibrosis transmembrane conductance regulator (CFTR) cellular processing in homozygous mutant mice, restoring nasal potential differences and improving survival (Egan, M. E., Pearson, M., Weiner, S. A., Rajendran, V., Rubin, D., Glockner-Pagel, J., Canny, S., Du, K., Lukacs, G. L., and Caplan, M. J. (2004) Science 304, 600-602). Because of the implied potential use of curcumin or similar compounds in the therapy of cystic fibrosis caused by the ΔF508 mutation, we tried to reproduce and extend the pre-clinical data of Egan et al. Fluorometric measurements of iodide influx in Fischer rat thyroid cells expressing ΔF508-CFTR showed no effect of curcumin (1-40 μM) when added for up to 24 h prior to assay in cells grown at 37 °C. Controls, including 27 °C rescue and 4 mM phenylbutyrate at 37 °C, were strongly positive. Also, curcumin did not increase short circuit current in primary cultures of a human airway epithelium homozygous for ΔF508-CFTR with a 27 °C rescue-positive control. Nasal potential differences in mice were measured in response to topical perfusion with serial solutions containing amiloride, low Cl-, and forskolin. Robust low Cl- and forskolin-induced hyperpolarization of 22 ± 3 mV was found in wild type mice, with 2.1 ± 0.4 mV hyperpolarization in ΔF508 homozygous mutant mice. No significant increase in Cl-/forskolin hyperpolarization was seen in any of the 22 ΔF508 mice studied using different curcumin preparations and administration regimens, including that used by Egan et al. Assay of serum curcumin by ethyl acetate extraction followed by liquid chromatography/mass spectrometry indicated a maximum serum concentration of 60 nM, well below that of 5-15 μM, where cellular effects by sarcoplasmic/endoplasmic reticulum calcium pump inhibition are proposed to occur. Our results do not support further evaluation of curcumin for cystic fibrosis therapy.

AB - Curcumin, the yellow colored component of the spice turmeric, has been reported to rescue defective ΔF508-cystic fibrosis transmembrane conductance regulator (CFTR) cellular processing in homozygous mutant mice, restoring nasal potential differences and improving survival (Egan, M. E., Pearson, M., Weiner, S. A., Rajendran, V., Rubin, D., Glockner-Pagel, J., Canny, S., Du, K., Lukacs, G. L., and Caplan, M. J. (2004) Science 304, 600-602). Because of the implied potential use of curcumin or similar compounds in the therapy of cystic fibrosis caused by the ΔF508 mutation, we tried to reproduce and extend the pre-clinical data of Egan et al. Fluorometric measurements of iodide influx in Fischer rat thyroid cells expressing ΔF508-CFTR showed no effect of curcumin (1-40 μM) when added for up to 24 h prior to assay in cells grown at 37 °C. Controls, including 27 °C rescue and 4 mM phenylbutyrate at 37 °C, were strongly positive. Also, curcumin did not increase short circuit current in primary cultures of a human airway epithelium homozygous for ΔF508-CFTR with a 27 °C rescue-positive control. Nasal potential differences in mice were measured in response to topical perfusion with serial solutions containing amiloride, low Cl-, and forskolin. Robust low Cl- and forskolin-induced hyperpolarization of 22 ± 3 mV was found in wild type mice, with 2.1 ± 0.4 mV hyperpolarization in ΔF508 homozygous mutant mice. No significant increase in Cl-/forskolin hyperpolarization was seen in any of the 22 ΔF508 mice studied using different curcumin preparations and administration regimens, including that used by Egan et al. Assay of serum curcumin by ethyl acetate extraction followed by liquid chromatography/mass spectrometry indicated a maximum serum concentration of 60 nM, well below that of 5-15 μM, where cellular effects by sarcoplasmic/endoplasmic reticulum calcium pump inhibition are proposed to occur. Our results do not support further evaluation of curcumin for cystic fibrosis therapy.

UR - http://www.scopus.com/inward/record.url?scp=4644360693&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4644360693&partnerID=8YFLogxK

U2 - 10.1074/jbc.M407308200

DO - 10.1074/jbc.M407308200

M3 - Article

C2 - 15280357

AN - SCOPUS:4644360693

VL - 279

SP - 40629

EP - 40633

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 39

ER -