Co-expression of mutated and normal adrenoleukodystrophy protein reduces protein function: implications for gene therapy of X-linked adrenoleukodystrophy
Abstract
Inherited defects in the X-chromosomal adrenoleukodystrophy (ALD; ABCD1) gene are the genetic cause of the severe neurodegenerative disorder X-linked adrenoleukodystrophy (X-ALD). Biochemically the accumulation of very long-chain fatty acids, caused by impaired peroxisomal beta-oxidation, is the pathognomonic characteristic of the disease. Due to the X-chromosomal inheritance of X-ALD no data are available to clarify the question whether mutated adrenoleukodystrophy proteins (ALDPs) can negatively influence normal ALDP function. Here we show that restoration of beta-oxidation in X-ALD fibroblasts following transient transfection with normal ALD cDNA is more effective in ALDP-deficient fibroblasts compared with fibroblasts expressing normal amounts of mutated ALDP. Furthermore, we utilized the HeLa Tet-on system to construct a stable HeLa cell line expressing a constant level of endogenous ALDP and doxycycline-inducible levels of mutated ALDP. The induction was doxycycline dosage-dependent and the ALDP correctly localized. Interestingly, although mutated ALDP increased >6-fold in a dosage-dependent manner the total amount of ALDP (mutated and normal) remained approximately even as demonstrated by western blot and flow cytometric analyses. Thus, apparently mutated and normal ALDP compete for integration into a limited number of sites in the peroxisomal membrane. Consequently, increased amounts of mutated ALDP resulted in decreased peroxisomal beta-oxidation and accumulation of very long-chain fatty acids. These findings have direct implications on future gene therapy approaches for treatment of X-ALD, since in some patients a non-functional endogenous protein could act in a dominant negative way or displace the introduced, normal protein.
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If a mutated ABCD1 protein dimerizes with a functional monomer, it might exert a dominant-negative effect, resulting in a nonfunctional transporter.
“If such a mutated ABCD1 protein dimerizes with a functional monomer, it might exert a dominant-negative effect on its dimerization partner, thus resulting in a nonfunctional transporter.84”
From The genetic landscape of X-linked adrenoleukodystrophy: inheritance, mutations, modifier genes, and diagnosis (2015) · cites this paper as [84]
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Cites
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
LAEMMLI, U. K. (1970) · Nature · doi:10.1038/227680a0 · Cited by 249,961 · see citing works
A SIMPLE METHOD FOR THE ISOLATION AND PURIFICATION OF TOTAL LIPIDES FROM ANIMAL TISSUES
Folch, J., Lees, M., & Stanley, G. H. S. (1957) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)64849-5 · Cited by 65,189 · see citing works
The Metabolic and Molecular Bases of Inherited Disease
Scriver, R. C. (1995) · Medical Entomology and Zoology · Cited by 12,101 · see citing works
Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
Thomas, P. S. (1980) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.77.9.5201 · Cited by 7,937 · see citing works
Transcriptional Activation by Tetracyclines in Mammalian Cells
Gossen, M., Freundlieb, S., Bender, G., Müller, G., Hillen, W., & Bujard, H. (1995) · Science · doi:10.1126/science.7792603 · Cited by 2,488 · cited 2× · see citing works
“For further investigation of the functional relationship between mutated and normal ALDP, we used the HeLa Tet-on system (15), which allows stringent control of mutant ALDP over a wide range of expression under an inducible promoter whereas endogenous ALDP levels remain constant.”
MethodPutative X-linked adrenoleukodystrophy gene shares unexpected homology with ABC transporters
Mosser, J., Douar, A.-M., Sarde, C.-O., Kioschis, P., Feil, R., Moser, H., Poustka, A.-M., Mandel, J.-L., & Aubourg, P. (1993) · Nature · doi:10.1038/361726a0 · Cited by 1,202 · see citing works
Moser, H. W., Mahmood, A., & Raymond, G. V. (2007) · Nature Clinical Practice Neurology · doi:10.1038/ncpneuro0421 · Cited by 663 · see citing works
Adrenoleukodystrophy: phenotype, genetics, pathogenesis and therapy
Moser, H. (1997) · Brain · doi:10.1093/brain/120.8.1485 · Cited by 478 · see citing works
Aubourg, P., Blanche, S., Jambaqué, I., Rocchiccioli, F., Kalifa, G., Naud-Saudreau, C., Rolland, M.-O., Debré, M., Chaussain, J.-L., Griscelli, C., et al. (1990) · New England Journal of Medicine · doi:10.1056/nejm199006283222607 · Cited by 366 · see citing works
Techniques in diagnostic human biochemical genetics
Shapira, E. (1991) · Europe PMC (PubMed Central) · Cited by 351 · see citing works
Migeon, B. R., Moser, H. W., Moser, A. B., Axelman, J., Sillence, D., & Norum, R. A. (1981) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.78.8.5066 · Cited by 283 · see citing works
Krivit, W. A., Peters, C., & Shapiro, E. (1999) · Current Opinion in Neurology · doi:10.1097/00019052-199904000-00007 · Cited by 275 · see citing works
Krivit, W., Sung, J. H., Shapiro, E., & Lockman, L. A. (1995) · Cell Transplantation · doi:10.1016/0963-6897(95)00021-o · Cited by 206 · see citing works
Bone marrow gene transfer in three patients with adenosine deaminase deficiency.
Hoogerbrugge, P. M., van Beusechem, V. W., Fischer, A., Debree, M., Deist, F. L., Pérignon, J. L., Morgan, G., Gaspar, B., Fairbanks, L. D., Skeoch, C. H., et al. (1996) · PubMed · Cited by 191 · see citing works
X-linked adrenoleukodystrophy: Genes, mutations, and phenotypes
Smith, K. D., Kemp, S., Braiterman, L. T., Lu, J.-F., Wei, H.-M., Geraghty, M., Stetten, G., Bergin, J. S., Pevsner, J., & Watkins, P. A. (1999) · Neurochemical Research · doi:10.1023/a:1022535930009 · Cited by 190 · see citing works
Firefly luciferase is targeted to peroxisomes in mammalian cells.
Keller, G. A., Gould, S. J., DeLuca, M., & Subramani, S. E. (1987) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.84.10.3264 · Cited by 188 · see citing works
Krivit, W. A., Lockman, L. A., Watkins, P. A., Hirsch, J., & Shapiro, E. (1995) · Journal of Inherited Metabolic Disease · doi:10.1007/bf00710052 · Cited by 159 · see citing works
Netik, A., Forss-Petter, S., Holzinger, A., Molzer, B., Unterrainer, G., & Berger, J. (1999) · Human Molecular Genetics · doi:10.1093/hmg/8.5.907 · Cited by 148 · see citing works
Self-citationHomo- and Heterodimerization of Peroxisomal ATP-binding Cassette Half-transporters
Liu, L. X., Janvier, K., Berteaux-Lecellier, V., Cartier, N., Benarous, R., & Aubourg, P. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.46.32738 · Cited by 141 · see citing works
Spectrum of mutations in the gene encoding the adrenoleukodystrophy protein.
(1995) · Europe PMC (PubMed Central) · Cited by 130 · see citing works
Altered expression of ALDP in X-linked adrenoleukodystrophy.
(1995) · PubMed Central · Cited by 127 · see citing works
Krivit, W. A., Aubourg, P. R., Shapiro, E., & Peters, C. (1999) · Current Opinion in Hematology · doi:10.1097/00062752-199911000-00004 · Cited by 124 · see citing works
Expression of the adrenoleukodystrophy protein in the human and mouse central nervous system
Fouquet, F., Zhou, J. M., Ralston, E., Murray, K., Troalen, F., Magal, E., Robain, O., Dubois-Dalcq, M., & Aubourg, P. (1997) · Neurobiology of Disease · doi:10.1006/nbdi.1997.0127 · Cited by 115 · see citing works
Bone marrow transplant in adrenoleukodystrophy
Moser, H. W., Tutschka, P. J., Brown, F. R., Moser, A. E., Yeager, A. M., Singh, I., Mark, S. A., Kumar, A., McDonnell, J. M., White, C. L., et al. (1984) · Neurology · doi:10.1212/wnl.34.11.1410 · Cited by 107 · see citing works
Cartier, N., Lopez, J., Moullier, P., Rocchiccioli, F., Rolland, M. O., Jorge, P., Mosser, J., Mandel, J. L., Bougnères, P. F., & Danos, O. (1995) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.92.5.1674 · Cited by 105 · see citing works
(1996) · PubMed Central · Cited by 99 · see citing works
Braiterman, L. (1998) · Human Molecular Genetics · doi:10.1093/hmg/7.2.239 · Cited by 98 · see citing works
Characterization of the 70-kDa Peroxisomal Membrane Protein, an ATP Binding Cassette Transporter
Imanaka, T., Aihara, K., Takano, T., Yamashita, A., Sato, R., Suzuki, Y., Yokota, S., & Osumi, T. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.17.11968 · Cited by 97 · see citing works
Moser, H. W. (1996) · Molecular and Chemical Neuropathology · doi:10.1007/bf02815041 · Cited by 96 · see citing works
Peroxisomal fatty acid β-oxidation in HepG2 cells
Watkins, P. A., Ferrell, E. V., Pedersen, J. I., & Hoefler, G. (1991) · Archives of Biochemistry and Biophysics · doi:10.1016/0003-9861(91)90419-j · Cited by 86 · see citing works
Takano, H., Koike, R., Onodera, O., Sasaki, R., & Tsuji, S. (1999) · Archives of Neurology · doi:10.1001/archneur.56.3.295 · Cited by 58 · see citing works
Missense mutations are frequent in the gene for X-chromosomal adrenoleukodystrophy (ALD)
Fuchs, S., Sarde, C. O., Wedemann, H., Schwinger, E., Mandel, J. L., & Gal, A. (1994) · Human Molecular Genetics · doi:10.1093/hmg/3.10.1903 · Cited by 57 · see citing works
Fairbairn, L., Lashford, L. S., Spooncer, E., McDermott, R. H., Lebens, G., Arrand, J. R., Arrand, J. R., Bellantuono, I., Holt, R. C. L., Hatton, C. E., et al. (1996) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.93.5.2025 · Cited by 50 · see citing works
Clinical and Genetic Aspects of X-Linked Adrenoleukodystrophy
Gärtner, J., Braun, A. D., Holzinger, A., Roerig, P., Lenard, H. G., & Roscher, A. (1998) · Neuropediatrics · doi:10.1055/s-2007-973526 · Cited by 50 · see citing works
Flavigny, E., Sanhaj, A., Aubourg, P. R., & Cartier, N. (1999) · FEBS Letters · doi:10.1016/s0014-5793(99)00379-8 · Cited by 49 · see citing works
Adrenoleukodystrophy: The Restoration of Peroxisomal β-Oxidaton by Transfection of Normal cDNA
Shinnoh, N., Yamada, T., Yoshimura, T., Furuya, H., Yoshida, Y., Suzuki, Y., Shimozawa, N., Orii, T., & Kobayashi, T. (1995) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1995.1733 · Cited by 43 · see citing works
Retroviral Transfer and Long-Term Expression of the Adrenoleukodystrophy Gene in Human CD34 + Cells
Doerflinger, N., Micléa, J.-M., Lopez, J., Chomienne, C., Bougneres, P. F., Aubourg, P. R., & Cartier, N. (1998) · Human Gene Therapy · doi:10.1089/hum.1998.9.7-1025 · Cited by 39 · see citing works
Treatment of adrenoleukodystrophy with bone marrow transplantation
Malm, G., Ringdén, O. T. H., Anvret, M., von Döbeln, U., Hagenfeldt, L., Isberg, B., Knuutila, S., Nennesmo, I., Winiarski, J. H., & Marcus, C. (1997) · Acta Paediatrica · doi:10.1111/j.1651-2227.1997.tb08918.x · Cited by 37 · see citing works
Mazurier, F., Moreau‐Gaudry, F., Salesse, S., Barbot, C., Ged, C., Reiffers, J. J., & de Verneuil, H. (1997) · Journal of Inherited Metabolic Disease · doi:10.1023/a:1005365008147 · Cited by 32 · see citing works
Wichers, M., Köhler, W., Brennemann, W., Boese, V., Sokolowski, P., BIDLINGMAIER, F., & Ludwig, M. Z. (1999) · Human Genetics · doi:10.1007/s004390051072 · Cited by 19 · see citing works
Forss-Petter, S. (1997) · doi:10.1002/(sici)1097-4547(19971201)50:5
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Berger, J., Wiesinger, C., & Eichler, F. (2015) · The Application of Clinical Genetics · doi:10.2147/tacg.s49590 · Cited by 140 · see citing works
Theodoulou, F. L., Holdsworth, M. J., & Baker, A. (2006) · FEBS Letters · doi:10.1016/j.febslet.2005.12.095 · Cited by 119 · see citing works
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Hsich, G. E., Sena‐Esteves, M., & Breakefield, X. O. (2002) · Human Gene Therapy · doi:10.1089/10430340252837198 · Cited by 98 · see citing works
Current and Future Pharmacological Treatment Strategies in X‐Linked Adrenoleukodystrophy
Berger, J., Pujol, A., Aubourg, P. R., & Forss‐Petter, S. (2010) · Brain Pathology · doi:10.1111/j.1750-3639.2010.00393.x · Cited by 90 · see citing works
Treatment of cerebral adrenoleukodystrophy: allogeneic transplantation and lentiviral gene therapy
Gupta, A. O., Raymond, G., Pierpont, E. I., Kemp, S., McIvor, R. S., Rayannavar, A., Miller, B., Lund, T. C., & Orchard, P. J. (2022) · Expert Opinion on Biological Therapy · doi:10.1080/14712598.2022.2124857 · Cited by 67 · see citing works
“limited, and that increased expression of wild-type ALD protein can influence the relative amount in the membrane as opposed to a defective gene product [109].”
BackgroundLeukodystrophies: recent developments in genetics, molecular biology, pathogenesis and treatment*
Berger, J., Moser, H. W., & Forss‐Petter, S. (2001) · Current Opinion in Neurology · doi:10.1097/00019052-200106000-00007 · Cited by 64 · see citing works
Benhamida, S., Pflumio, F., Dubart-Kupperschmitt, A., Zhao-Emonet, J.-C., Cavazzana-Calvo, M., Rocchiccioli, F., Fichelson, S., Aubourg, P., Charneau, P., & Cartier, N. (2003) · Molecular Therapy · doi:10.1016/s1525-0016(03)00002-9 · Cited by 61 · see citing works
“Some authors [26] have raised concerns about the ability to correct ALD cells with missense mutations and present although nonfunctional ALD protein, due to competition for insertion or function within the peroxisomal membrane of normal and mutant forms of ALD protein.”
BackgroundWeinhofer, I. (2002) · Human Molecular Genetics · doi:10.1093/hmg/11.22.2701 · Cited by 59 · see citing works
Genin, E. C., Geillon, F., Gondcaille, C., Athias, A., Gambert, P., Trompier, D., & Savary, S. (2011) · Journal of Biological Chemistry · doi:10.1074/jbc.m110.211912 · Cited by 53 · see citing works
Zhang, X., De Marcos Lousa, C., Schutte-Lensink, N., Ofman, R., Wanders, R. J., Baldwin, S. A., Baker, A., Kemp, S., & Theodoulou, F. L. (2011) · Biochemical Journal · doi:10.1042/bj20110249 · Cited by 43 · see citing works
Cytotoxic effect of doxycycline and its implications for tet-on gene expression systems
Ermak, G., Cancasci, V. J., & Davies, K. J. A. (2003) · Analytical Biochemistry · doi:10.1016/s0003-2697(03)00166-0 · Cited by 40 · see citing works
The mitochondrial ABC transporter Atm1p functions as a homodimer
Chloupková, M., Reaves, S. K., LeBard, L. M., & Koeller, D. M. (2004) · FEBS Letters · doi:10.1016/j.febslet.2004.05.051 · Cited by 27 · see citing works
ABCD1 translation–initiator mutation demonstrates genotype–phenotype correlation for AMN
O’Neill, G. N., Aoki, M., & Brown, R. H. (2001) · Neurology · doi:10.1212/wnl.57.11.1956 · Cited by 28 · see citing works
Maya‐Mendoza, A., & Aranda‐Anzaldo, A. (2003) · Analytical Biochemistry · doi:10.1016/s0003-2697(02)00611-5 · Cited by 24 · see citing works
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Kálmán, B., & Leist, T. P. (2004) · The Neurologist · doi:10.1097/01.nrl.0000131273.12774.16 · Cited by 18 · see citing works
Existence of catalase-less peroxisomes in Sf21 insect cells
Kurisu, M., Morita, M., Kashiwayama, Y., Yokota, S., Hayashi, H., Sakai, Y., Ohkuma, S., Nishimura, M., & Imanaka, T. (2003) · Biochemical and Biophysical Research Communications · doi:10.1016/s0006-291x(03)00913-6 · Cited by 16 · see citing works
Raymond, G. V. (2001) · Current Opinion in Neurology · doi:10.1097/00019052-200112000-00017 · Cited by 12 · see citing works
Zuo, X., & Chen, Z. (2024) · Lipids in Health and Disease · doi:10.1186/s12944-024-02361-0 · Cited by 12 · see citing works
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Gueugnon, F., Volodina, N., Taouil, J. E., Lopez, T. E., Gondcaille, C., Grand, A. S.-L., Mooijer, P. A. W., Kemp, S., Wanders, R. J. A., & Savary, S. (2006) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2005.12.152 · Cited by 9 · see citing works
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Kozlova, A. M., Morshneva, A. V., Gnedina, O. O., & Igotti, M. V. (2025) · Cell and Tissue Biology · doi:10.1134/s1990519x24600777 · Cited by 1 · see citing works
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Patricio, S. T., de la Vega, J. A. B., Álvarez, D. N., GonzáLez, F. J. G., Fernández, M. D. L., & Hachero, J. G. (2009) · Endocrinología y Nutrición · doi:10.1016/s1575-0922(09)70192-5 · Cited by 1 · see citing works
Lack, E. E., & Wieneke, J. A. (2008) · Elsevier eBooks · doi:10.1016/b978-0-323-01970-5.50018-x
Peña, S. O., & Barrera, A. (2005)
Lack, E. E., & Paal, E. E. (2020) · Elsevier eBooks · doi:10.1016/b978-0-323-54941-7.00016-5
Shi, X., Qi, X., Zheng, J., Ma, J., & Li, D. (2024) · Frontiers in Genetics · doi:10.3389/fgene.2024.1421122
Lack, E. E., & Paal, E. E. (2014) · Elsevier eBooks · doi:10.1016/b978-1-4557-4327-8.00016-4
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- Authors
- Unterrainer, G.
- Affiliations
- University of Vienna (AT)
- Corresponding author
- Unterrainer, G.
- Venue
- Human Molecular Genetics
- Publisher
- Oxford University Press (OUP)
- Published
- 2000-11-01
- Volume
- 9
- Issue
- 18
- Pages
- 2609-2616
- ISSN
- 0964-6906 (ISSN-L)
- 1460-2083
- OpenAlex
- W2120211298
- Cited by
- 30
- References
- 41
- Citations by year
- 2025: 1 · 2024: 2 · 2022: 2 · 2020: 1 · 2017: 1 · 2015: 1 · 2014: 1
- FWCI
- 1.64
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Human Molecular Genetics (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Metabolism and Genetic Disorders
- Health Sciences › Medicine › Physiology › Lysosomal Storage Disorders Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- X-linked adrenoleukodystrophy, ABCD1, dominant negative effect, protein co-expression, peroxisomal beta-oxidation, very long chain fatty acids, gene therapy, doxycycline-inducible expression, fibroblasts
- MeSH terms
- ATP Binding Cassette Transporter, Subfamily D, Member 1, Adrenoleukodystrophy, Doxycycline, Fatty Acids, Female, Fibroblasts, Flow Cytometry, HeLa Cells, Humans, Genetic Linkage, Male, Membrane Proteins, Molecular Sequence Data, Oxidation-Reduction, Protein Conformation, RNA, Messenger, Transfection, X Chromosome, Blotting, Western, Genetic Therapy, Gene Expression, Point Mutation, ATP-Binding Cassette Transporters, Fluorescent Antibody Technique, Indirect, Peroxisomes
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