6 research outputs found
Investigating inherited renal disorders and identification of a novel cause of Joubert syndrome
MDThesisJoubert syndrome (JBTS) is a genetically heterogeneous neurodevelopmental
ciliopathy that can have severe renal manifestations requiring renal replacement
therapy. JBTS can be caused by mutation in 34 genes, however more than 50% of
JBTS cases have unknown genetic causes. Through a multidisciplinary renalgenetics family clinic we studied a cohort of patients with inherited renal disorders
and identified two families with JBTS phenotype and unknown molecular genetic
diagnosis. We further investigated the underlying genetic aetiology of Joubert
syndrome using combined homozygosity mapping of both families highlighted a
candidate locus on chromosome 10, and whole exome sequencing revealed two
missense variants in ARL3 within the candidate locus. The encoded protein, ADP
ribosylation factor-like GTPase 3 (ARL3), is a small GTP-binding protein that is
involved in trafficking lipid-modified proteins into the cilium in a GTP-dependent
manner. Both missense variants replace the highly conserved Arg149 residue, which
we show to be necessary for the interaction with its guanine nucleotide exchange
factor ARL13B. Using patients derived fibroblasts, we identified that the mutant
ARL3 protein is associated with reduced ciliary cargo protein INPP5E and NPHP3
localization in cilia
ARL3 mutations cause Joubert syndrome by disrupting ciliary protein composition
Joubert syndrome (JBTS) is a genetically heterogeneous autosomal recessive neurodevelopmental
ciliopathy. We investigated further the underlying genetic etiology of Joubert syndrome by studying
two unrelated families in whom JBTS was not associated with pathogenic variants in known JBTSrelated
genes. Combined autozygosity mapping of both families highlighted a candidate locus on
chromosome 10 (chr10: 101569997-109106128 (hg 19)), and exome sequencing revealed two
missense variants in ARL3 within the candidate locus. The encoded protein, ADP Ribosylation
Factor-Like GTPase 3, ARL3, is a small GTP-binding protein that is involved in directing lipid-modified
proteins into the cilium in a GTP-dependent manner. Both missense variants replace the highly
conserved Arg149 residue, which we show to be necessary for the interaction with its guanine
nucleotide exchange factor ARL13B, such that the mutant protein is associated with reduced INPP5E
and NPHP3 localisation in cilia. We propose that ARL3 provides a potential hub in the network of
encoded ciliopathy genes, whereby perturbation of ARL3 results in the mislocalisation of multiple
ciliary proteins due to abnormal displacement of lipidated protein cargo
Targeted exon skipping of a CEP290 mutation rescues Joubert syndrome phenotypes in vitro and in a murine model
International audienceGenetic treatments of renal ciliopathies leading to cystic kidney disease would provide a real advance in current therapies. Mutations in CEP290 underlie a ciliopathy called Joubert syndrome (JBTS). Human disease phenotypes include cerebral, retinal, and renal disease, which typically progresses to end stage renal failure (ESRF) within the first two decades of life. While currently incurable, there is often a period of years between diagnosis and ESRF that provides a potential window for therapeutic intervention. By studying patient biopsies, patient-derived kidney cells, and a mouse model, we identify abnormal elongation of primary cilia as a key pathophysiological feature of CEP290-associated JBTS and show that antisense oligonucleotide (ASO)-induced splicing of the mutated exon (41, G1890*) restores protein expression in patient cells. We demonstrate that ASO-induced splicing leading to exon skipping is tolerated, resulting in correct localization of CEP290 protein to the ciliary transition zone, and restoration of normal cilia length in patient kidney cells. Using a gene trap Cep290 mouse model of JBTS, we show that systemic ASO treatment can reduce the cystic burden of diseased kidneys in vivo. These findings indicate that ASO treatment may represent a promising therapeutic approach for kidney disease in CEP290-associated ciliopathy syndromes
Acidosis and Deafness in Patients with Recessive Mutations in FOXI1
Maintenance of the composition of inner ear fluid and regulation of electrolytes and acid-base homeostasis in the collecting duct system of the kidney require an overlapping set of membrane transport proteins regulated by the forkhead transcription factor FOXI1. In two unrelated consanguineous families, we identified three patients with novel homozygous missense mutations in FOXI1 (p.L146F and p.R213P) predicted to affect the highly conserved DNA binding domain. Patients presented with early-onset sensorineural deafness and distal renal tubular acidosis. In cultured cells, the mutations reduced the DNA binding affinity of FOXI1, which hence, failed to adequately activate genes crucial for normal inner ear function and acid-base regulation in the kidney. A substantial proportion of patients with a clinical diagnosis of inherited distal renal tubular acidosis has no identified causative mutations in currently known disease genes. Our data suggest that recessive mutations in FOXI1 can explain the disease in a subset of these patients
Early Diagnosis of Classic Homocystinuria in Kuwait through Newborn Screening: A 6-Year Experience
Kuwait is a small Arabian Gulf country with a high rate of consanguinity and where a national newborn screening program was expanded in October 2014 to include a wide range of endocrine and metabolic disorders. A retrospective study conducted between January 2015 and December 2020 revealed a total of 304,086 newborns have been screened in Kuwait. Six newborns were diagnosed with classic homocystinuria with an incidence of 1:50,000, which is not as high as in Qatar but higher than the global incidence. Molecular testing for five of them has revealed three previously reported pathogenic variants in the CBS gene, c.969G>A, p.(Trp323Ter); c.982G>A, p.(Asp328Asn); and the Qatari founder variant c.1006C>T, p.(Arg336Cys). This is the first study to review the screening of newborns in Kuwait for classic homocystinuria, starting with the detection of elevated blood methionine and providing a follow-up strategy for positive results, including plasma total homocysteine and amino acid analyses. Further, we have demonstrated an increase in the specificity of the current newborn screening test for classic homocystinuria by including the methionine to phenylalanine ratio along with the elevated methionine blood levels in first-tier testing. Here, we provide evidence that the newborn screening in Kuwait has led to the early detection of classic homocystinuria cases and enabled the affected individuals to lead active and productive lives