Expression and In Vivo rescue of human ABCC6 disease-causing mutants in mouse liver

Olivier Le Saux, Krisztina Fülöp, Yukiko Yamaguchi, Attila Iliás, Z. Szabó, Christopher N. Brampton, Viola Pomozi, Krisztina Huszár, T. Arányi, A. Váradi

Research output: Contribution to journalArticle

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Abstract

Loss-of-function mutations in ABCC6 can cause chronic or acute forms of dystrophic mineralization described in disease models such as pseudoxanthoma elasticum (OMIM 26480) in human and dystrophic cardiac calcification in mice. The ABCC6 protein is a large membrane-embedded organic anion transporter primarily found in the plasma membrane of hepatocytes. We have established a complex experimental strategy to determine the structural and functional consequences of disease-causing mutations in the human ABCC6. The major aim of our study was to identify mutants with preserved transport activity but failure in intracellular targeting. Five missense mutations were investigated: R1138Q, V1298F, R1314W, G1321S and R1339C. Using in vitro assays, we have identified two variants; R1138Q and R1314W that retained significant transport activity. All mutants were transiently expressed in vivo, in mouse liver via hydrodynamic tail vein injections. The inactive V1298F was the only mutant that showed normal cellular localization in liver hepatocytes while the other mutants showed mostly intracellular accumulation indicating abnormal trafficking. As both R1138Q and R1314W displayed endoplasmic reticulum localization, we tested whether 4-phenylbutyrate (4-PBA), a drug approved for clinical use, could restore their intracellular trafficking to the plasma membrane in MDCKII and mouse liver. The cellular localization of R1314W was significantly improved by 4-PBA treatment, thus potentially rescuing its physiological function. Our work demonstrates the feasibility of the in vivo rescue of cellular maturation of some ABCC6 mutants in physiological conditions very similar to the biology of the fully differentiated human liver and could have future human therapeutic application.

Original languageEnglish
Article numbere24738
JournalPLoS One
Volume6
Issue number9
DOIs
Publication statusPublished - Sep 14 2011

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human diseases
Liver
mutants
liver
mice
Cell membranes
Hepatocytes
hepatocytes
Cell Membrane
Pseudoxanthoma Elasticum
Organic Anion Transporters
Genetic Databases
plasma membrane
Mutation
Rubber
Missense Mutation
Hydrodynamics
missense mutation
Endoplasmic Reticulum
disease models

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Expression and In Vivo rescue of human ABCC6 disease-causing mutants in mouse liver. / Le Saux, Olivier; Fülöp, Krisztina; Yamaguchi, Yukiko; Iliás, Attila; Szabó, Z.; Brampton, Christopher N.; Pomozi, Viola; Huszár, Krisztina; Arányi, T.; Váradi, A.

In: PLoS One, Vol. 6, No. 9, e24738, 14.09.2011.

Research output: Contribution to journalArticle

Le Saux, O, Fülöp, K, Yamaguchi, Y, Iliás, A, Szabó, Z, Brampton, CN, Pomozi, V, Huszár, K, Arányi, T & Váradi, A 2011, 'Expression and In Vivo rescue of human ABCC6 disease-causing mutants in mouse liver', PLoS One, vol. 6, no. 9, e24738. https://doi.org/10.1371/journal.pone.0024738
Le Saux, Olivier ; Fülöp, Krisztina ; Yamaguchi, Yukiko ; Iliás, Attila ; Szabó, Z. ; Brampton, Christopher N. ; Pomozi, Viola ; Huszár, Krisztina ; Arányi, T. ; Váradi, A. / Expression and In Vivo rescue of human ABCC6 disease-causing mutants in mouse liver. In: PLoS One. 2011 ; Vol. 6, No. 9.
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