Abstract
Cystinosis is an autosomal recessive disorder characterized by an accumulation of intralysosomal cystine. The causative gene, CTNS, encodes cystinosin, a seven-transmembrane-domain protein, which we recently showed to be a lysosomal cystine transporter. The most severe and frequent form of cystinosis, the infantile form, appears around 6 to 12 months, with a proximal tubulopathy (de Toni-Debré-Fanconi syndrome) and ocular damage. End-stage renal failure is reached by 10 years of age. Accumulation of cystine in all tissues eventually leads to multisystemic disease. Treatment with cysteamine, which reduces the concentration of intracellular cystine, delays disease progression but has undesirable side effects. We report the first Ctns knockout mouse model generated using a promoter trap approach. We replaced the last four Ctns exons by an internal ribosome entry site-betagal-neo cassette and showed that the truncated protein was mislocalized and nonfunctional. Ctns(-/-) mice accumulated cystine in all organs tested, and cystine crystals, pathognomonic of cystinosis, were observed. Ctns(-/-) mice developed ocular changes similar to those observed in affected individuals, bone defects and behavioral anomalies. Interestingly, Ctns(-/-) mice did not develop signs of a proximal tubulopathy, or renal failure. A preliminary therapeutic trial using an oral administration of cysteamine was carried out and demonstrated the efficiency of this treatment for cystine clearance in Ctns(-/-) mice. This animal model will prove an invaluable and unique tool for testing emerging therapeutics for cystinosis.
MeSH Terms
Alleles
Amino Acid Transport Systems, Neutral
Animals
Bone and Bones/diagnostic imaging,pathology
Cystine/metabolism
Cystinosis/diagnostic imaging,genetics,metabolism
Cytosine/metabolism
Dogs
Electroretinography
Glycoproteins
Lysosomes/metabolism
Membrane Proteins/genetics,physiology
Membrane Transport Proteins
Mice
Mice, Inbred C57BL
Mice, Knockout
Models, Genetic
Promoter Regions, Genetic
Radiography
Recombinant Proteins/metabolism
Retina/abnormalities,pathology
Reverse Transcriptase Polymerase Chain Reaction
Time Factors
Tissue Distribution
Chemicals
Amino Acid Transport Systems, Neutral
CTNS protein, human
Glycoproteins
Membrane Proteins
Membrane Transport Proteins
Recombinant Proteins
Cystine
Cytosine
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Cherqui Stéphanie
INSERM U423, Hôpital Necker-Enfants Malades, Paris, France.
Sevin Caroline
Hamard Ghislaine
Kalatzis Vasiliki
Sich Mireille
Pequignot Marie O
Gogat Karïn
Abitbol Marc
Broyer Michel
Gubler Marie-Claire
Antignac Corinne
References (33)
33 references, click to expand
-
Glucose-dependent liver gene expression in upstream stimulatory factor 2 -/- mice.
J Biol Chem. 1997 Aug 29;272(35):21944-9
PMID: 9268329
-
The targeting of cystinosin to the lysosomal membrane requires a tyrosine-based signal and a novel sorting motif.
J Biol Chem. 2001 Apr 20;276(16):13314-21
PMID: 11150305
-
Generalized glycogen storage and cardiomegaly in a knockout mouse model of Pompe disease.
Hum Mol Genet. 1998 Jan;7(1):53-62
PMID: 9384603
-
The Fanconi syndrome with cystinossis. Electron microscopy of renal biopsy specimens from five patients.
Am J Med. 1962 Dec;33:893-910
PMID: 13957207
-
Parenchymal organ cystine depletion with long-term cysteamine therapy.
Biochem Med Metab Biol. 1992 Dec;48(3):275-85
PMID: 1476793
-
Clinical polymorphism of cystinosis encephalopathy. Results of treatment with cysteamine.
J Inherit Metab Dis. 1996;19(1):65-75
PMID: 8830179
-
Characterization of the lysosomal cystine transport system in mouse L-929 fibroblasts.
J Biol Chem. 1990 Jun 15;265(17):9888-95
PMID: 2141024
-
Ocular changes in long-term evolution of infantile cystinosis.
Ophthalmic Paediatr Genet. 1987 Jun;8(2):131-7
PMID: 3658340
-
Functional consequences of oncogene-induced horizontal cell degeneration in the retinas of transgenic mice.
Vis Neurosci. 1997 Jul-Aug;14(4):627-32
PMID: 9278992
-
Mutations of CTNS causing intermediate cystinosis.
Mol Genet Metab. 1999 Aug;67(4):283-93
PMID: 10444339
-
Binding assays for amino acids. The utilization of a cystine binding protein from Escherichia coli for the determination of acid-soluble cystine in small physiological samples.
J Biol Chem. 1974 Oct 10;249(19):6033-9
PMID: 4214006
-
Cystinosin, the protein defective in cystinosis, is a H(+)-driven lysosomal cystine transporter.
EMBO J. 2001 Nov 1;20(21):5940-9
PMID: 11689434
-
Severity of phenotype in cystinosis varies with mutations in the CTNS gene: predicted effect on the model of cystinosin.
Hum Mol Genet. 1999 Dec;8(13):2507-14
PMID: 10556299
-
Lack of complementation in somatic cell hybrids between fibroblasts from patients with different forms of cystinosis.
Proc Natl Acad Sci U S A. 1988 May;85(10):3531-4
PMID: 3368464
-
Increased cystine in leukocytes from individuals homozygous and heterozygous for cystinosis.
Science. 1967 Sep 15;157(3794):1321-2
PMID: 6038997
-
Germ line chimeras from female ES cells.
Exp Cell Res. 1997 Jan 10;230(1):45-9
PMID: 9013705
-
A detailed ethological analysis of the mouse open field test: effects of diazepam, chlordiazepoxide and an extremely low frequency pulsed magnetic field.
Neurosci Biobehav Rev. 2001 May;25(3):235-60
PMID: 11378179
-
A novel gene encoding an integral membrane protein is mutated in nephropathic cystinosis.
Nat Genet. 1998 Apr;18(4):319-24
PMID: 9537412
-
Myopathy and cystine storage in muscles in a patient with nephropathic cystinosis.
N Engl J Med. 1988 Dec 1;319(22):1461-4
PMID: 3185663
-
Cystinosis. An ultrastructural and electron-probe study of the kidney with unusual findings.
Arch Pathol. 1971 Mar;91(3):206-21
PMID: 4322430
-
Ocular nonnephropathic cystinosis: clinical, biochemical, and molecular correlations.
Pediatr Res. 2000 Jan;47(1):17-23
PMID: 10625078
-
Cysteamine therapy for children with nephropathic cystinosis.
N Engl J Med. 1987 Apr 16;316(16):971-7
PMID: 3550461
-
Distal myopathy in nephropathic cystinosis.
Pediatr Nephrol. 2000 Jan;14(1):36-8
PMID: 10654328
-
Cystine transport is defective in isolated leukocyte lysosomes from patients with cystinosis.
Science. 1982 Sep 24;217(4566):1263-5
PMID: 7112129
-
Role of adenosine triphosphate (ATP) and NaK ATPase in the inhibition of proximal tubule transport with intracellular cystine loading.
J Clin Invest. 1991 Mar;87(3):955-61
PMID: 1847941
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
Cytokines in Gaucher's disease.
Eur Cytokine Netw. 1999 Jun;10(2):205-10
PMID: 10400826
-
[Cystinosis with involvement of the central nervous system and muscles (myopathy)].
Zentralbl Allg Pathol. 1971;114(4):493-8
PMID: 5288177
-
Identification and characterisation of the murine homologue of the gene responsible for cystinosis, Ctns.
BMC Genomics. 2000;1:2
PMID: 11121245
-
Early development of the renal lesions in infantile cystinosis.
Pediatr Nephrol. 2000 Nov;15(1-2):50-6
PMID: 11095011
-
Infantile cystinosis: a reappraisal of early and late symptoms.
Adv Nephrol Necker Hosp. 1981;10:137-66
PMID: 6791467
-
Open-field behavior in muroid rodents.
Behav Biol. 1976 Aug;17(4):495-506
PMID: 788698
-
Ophthalmologic involvement in inherited renal disease.
Adv Nephrol Necker Hosp. 1992;21:143-56
PMID: 1566644