Compounds that correct F508del-CFTR trafficking can also correct other protein trafficking diseases: An in vitro study using cell lines

Heidi M. Sampson, Hung Lam, Pei Chun Chen, Donglei Zhang, Cristina Mottillo, Myriam Mirza, Karim Qasim, Alvin Shrier, Show-Ling Shyng, John W. Hanrahan, David Y. Thomas

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: Many genetic diseases are due to defects in protein trafficking where the mutant protein is recognized by the quality control systems, retained in the endoplasmic reticulum (ER), and degraded by the proteasome. In many cases, the mutant protein retains function if it can be trafficked to its proper cellular location. We have identified structurally diverse correctors that restore the trafficking and function of the most common mutation causing cystic fibrosis, F508del-CFTR. Most of these correctors do not act directly as ligands of CFTR, but indirectly on other pathways to promote folding and correction. We hypothesize that these proteostasis regulators may also correct other protein trafficking diseases. Methods: To test our hypothesis, we used stable cell lines or transient transfection to express 2 well-studied trafficking disease mutations in each of 3 different proteins: the arginine-vasopressin receptor 2 (AVPR2, also known as V2R), the human ether-a-go-go-related gene (KCNH2, also known as hERG), and finally the sulfonylurea receptor 1 (ABCC8, also known as SUR1). We treated cells expressing these mutant proteins with 9 structurally diverse F508del-CFTR correctors that function through different cellular mechanisms and assessed whether correction occurred via immunoblotting and functional assays. Results were deemed significantly different from controls by a one-way ANOVA (p <0.05). Results: Here we show that F508del-CFTR correctors RDR1, KM60 and KM57 also correct some mutant alleles of other protein trafficking diseases. We also show that one corrector, the cardiac glycoside ouabain, was found to alter the glycosylation of all mutant alleles tested. Conclusions: Correctors of F508del-CFTR trafficking might have broader applications to other protein trafficking diseases.

Original languageEnglish (US)
Pages (from-to)11
Number of pages1
JournalOrphanet Journal of Rare Diseases
DOIs
StateAccepted/In press - Jan 14 2013
Externally publishedYes

Fingerprint

Protein Transport
Mutant Proteins
Cell Line
Alleles
Sulfonylurea Receptors
Vasopressin Receptors
Cardiac Glycosides
Mutation
Inborn Genetic Diseases
Ouabain
Proteasome Endopeptidase Complex
Glycosylation
Immunoblotting
Cystic Fibrosis
Endoplasmic Reticulum
Quality Control
Ether
Transfection
Analysis of Variance
Ligands

ASJC Scopus subject areas

  • Pharmacology (medical)
  • Genetics(clinical)
  • Medicine(all)

Cite this

Compounds that correct F508del-CFTR trafficking can also correct other protein trafficking diseases : An in vitro study using cell lines. / Sampson, Heidi M.; Lam, Hung; Chen, Pei Chun; Zhang, Donglei; Mottillo, Cristina; Mirza, Myriam; Qasim, Karim; Shrier, Alvin; Shyng, Show-Ling; Hanrahan, John W.; Thomas, David Y.

In: Orphanet Journal of Rare Diseases, 14.01.2013, p. 11.

Research output: Contribution to journalArticle

Sampson, Heidi M. ; Lam, Hung ; Chen, Pei Chun ; Zhang, Donglei ; Mottillo, Cristina ; Mirza, Myriam ; Qasim, Karim ; Shrier, Alvin ; Shyng, Show-Ling ; Hanrahan, John W. ; Thomas, David Y. / Compounds that correct F508del-CFTR trafficking can also correct other protein trafficking diseases : An in vitro study using cell lines. In: Orphanet Journal of Rare Diseases. 2013 ; pp. 11.
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T2 - An in vitro study using cell lines

AU - Sampson, Heidi M.

AU - Lam, Hung

AU - Chen, Pei Chun

AU - Zhang, Donglei

AU - Mottillo, Cristina

AU - Mirza, Myriam

AU - Qasim, Karim

AU - Shrier, Alvin

AU - Shyng, Show-Ling

AU - Hanrahan, John W.

AU - Thomas, David Y.

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AB - Background: Many genetic diseases are due to defects in protein trafficking where the mutant protein is recognized by the quality control systems, retained in the endoplasmic reticulum (ER), and degraded by the proteasome. In many cases, the mutant protein retains function if it can be trafficked to its proper cellular location. We have identified structurally diverse correctors that restore the trafficking and function of the most common mutation causing cystic fibrosis, F508del-CFTR. Most of these correctors do not act directly as ligands of CFTR, but indirectly on other pathways to promote folding and correction. We hypothesize that these proteostasis regulators may also correct other protein trafficking diseases. Methods: To test our hypothesis, we used stable cell lines or transient transfection to express 2 well-studied trafficking disease mutations in each of 3 different proteins: the arginine-vasopressin receptor 2 (AVPR2, also known as V2R), the human ether-a-go-go-related gene (KCNH2, also known as hERG), and finally the sulfonylurea receptor 1 (ABCC8, also known as SUR1). We treated cells expressing these mutant proteins with 9 structurally diverse F508del-CFTR correctors that function through different cellular mechanisms and assessed whether correction occurred via immunoblotting and functional assays. Results were deemed significantly different from controls by a one-way ANOVA (p <0.05). Results: Here we show that F508del-CFTR correctors RDR1, KM60 and KM57 also correct some mutant alleles of other protein trafficking diseases. We also show that one corrector, the cardiac glycoside ouabain, was found to alter the glycosylation of all mutant alleles tested. Conclusions: Correctors of F508del-CFTR trafficking might have broader applications to other protein trafficking diseases.

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