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1.
Mol Genet Genomic Med ; 11(12): e2256, 2023 Dec.
Article de Anglais | MEDLINE | ID: mdl-37592902

RÉSUMÉ

BACKGROUND: Very long-chain fatty acids (VLCFAs) composed of more than 20 carbon atoms are essential in the biosynthesis of cell membranes in the brain, skin, and retina. VLCFAs are elongated beyond 28 carbon atoms by ELOVL4 enzyme. Variants in ELOVL4 are associated with three Mendelian disorders: autosomal dominant (AD) Stargardt-like macular dystrophy type 3, AD spinocerebellar ataxia, and autosomal recessive disorder congenital ichthyosis, spastic quadriplegia and impaired intellectual development (ISQMR). Only seven subjects from five unrelated families with ISQMR have been described, all of which have biallelic single-nucleotide variants. METHODS: We performed clinical exome sequencing on probands from four unrelated families with neuro-ichthyosis. RESULTS: We identified three novel homozygous ELOVL4 variants. Two of the families originated from the same Saudi tribe and had the exact homozygous exonic deletion in ELOVL4, while the third and fourth probands had two different novel homozygous missense variants. Seven out of the eight affected subjects had profound developmental delay, epilepsy, axial hypotonia, peripheral hypertonia, and ichthyosis. Delayed myelination and corpus callosum hypoplasia were seen in two of five subjects with brain magnetic rosonance imaging and cerebral atrophy in three. CONCLUSION: Our study expands the allelic spectrum of ELOVL4-related ISQMR. The detection of the same exonic deletion in two unrelated Saudi family from same tribe suggests a tribal founder mutation.


Sujet(s)
Ichtyose lamellaire , Ichtyose , Dégénérescence maculaire , Humains , Mutation , Dégénérescence maculaire/génétique , Rétine/métabolisme , Ichtyose/génétique , Carbone , Protéines de l'oeil/génétique , Protéines membranaires/génétique
3.
Genet Med ; 25(2): 100323, 2023 02.
Article de Anglais | MEDLINE | ID: mdl-36401616

RÉSUMÉ

PURPOSE: Pathogenic variants in genes encoding ubiquitin E3 ligases are known to cause neurodevelopmental syndromes. Additional neurodevelopmental disorders associated with the other genes encoding E3 ligases are yet to be identified. METHODS: Chromosomal analysis and exome sequencing were used to identify the genetic causes in 10 patients from 7 unrelated families with syndromic neurodevelopmental, seizure, and movement disorders and neurobehavioral phenotypes. RESULTS: In total, 4 patients were found to have 3 different homozygous loss-of-function (LoF) variants, and 3 patients had 4 compound heterozygous missense variants in the candidate E3 ligase gene, HECTD4, that were rare, absent from controls as homozygous, and predicted to be deleterious in silico. In 3 patients from 2 families with Angelman-like syndrome, paralog-directed candidate gene approach detected 2 LoF variants in the other candidate E3 ligase gene, UBE3C, a paralog of the Angelman syndrome E3 ligase gene, UBE3A. The RNA studies in 4 patients with LoF variants in HECTD4 and UBE3C provided evidence for the LoF effect. CONCLUSION: HECTD4 and UBE3C are novel biallelic rare disease genes, expand the association of the other HECT E3 ligase group with neurodevelopmental syndromes, and could explain some of the missing heritability in patients with a suggestive clinical diagnosis of Angelman syndrome.


Sujet(s)
Syndrome d'Angelman , Troubles du développement neurologique , Humains , Syndrome d'Angelman/génétique , Ubiquitine/génétique , Ubiquitin-protein ligases/génétique , Troubles du développement neurologique/génétique , Phénotype
4.
Hum Genet ; 142(3): 379-397, 2023 Mar.
Article de Anglais | MEDLINE | ID: mdl-36538041

RÉSUMÉ

CLEC16A is a membrane-associated C-type lectin protein that functions as a E3-ubiquitin ligase. CLEC16A regulates autophagy and mitophagy, and reportedly localizes to late endosomes. GWAS studies have associated CLEC16A SNPs to various auto-immune and neurological disorders, including multiple sclerosis and Parkinson disease. Studies in mouse models imply a role for CLEC16A in neurodegeneration. We identified bi-allelic CLEC16A truncating variants in siblings from unrelated families presenting with a severe neurodevelopmental disorder including microcephaly, brain atrophy, corpus callosum dysgenesis, and growth retardation. To understand the function of CLEC16A in neurodevelopment we used in vitro models and zebrafish embryos. We observed CLEC16A localization to early endosomes in HEK293T cells. Mass spectrometry of human CLEC16A showed interaction with endosomal retromer complex subunits and the endosomal ubiquitin ligase TRIM27. Expression of the human variant leading to C-terminal truncated CLEC16A, abolishes both its endosomal localization and interaction with TRIM27, suggesting a loss-of-function effect. CLEC16A knockdown increased TRIM27 adhesion to early endosomes and abnormal accumulation of endosomal F-actin, a sign of disrupted vesicle sorting. Mutagenesis of clec16a by CRISPR-Cas9 in zebrafish embryos resulted in accumulated acidic/phagolysosome compartments, in neurons and microglia, and dysregulated mitophagy. The autophagocytic phenotype was rescued by wild-type human CLEC16A but not the C-terminal truncated CLEC16A. Our results demonstrate that CLEC16A closely interacts with retromer components and regulates endosomal fate by fine-tuning levels of TRIM27 and polymerized F-actin on the endosome surface. Dysregulation of CLEC16A-mediated endosomal sorting is associated with neurodegeneration, but it also causes accumulation of autophagosomes and unhealthy mitochondria during brain development.


Sujet(s)
Actines , Danio zébré , Animaux , Humains , Protéines de liaison à l'ADN/métabolisme , Endosomes/génétique , Endosomes/métabolisme , Cellules HEK293 , Lectines de type C/génétique , Lectines de type C/composition chimique , Lectines de type C/métabolisme , Protéines membranaires/métabolisme , Transporteurs de monosaccharides/composition chimique , Transporteurs de monosaccharides/génétique , Transporteurs de monosaccharides/métabolisme , Protéines nucléaires/métabolisme , Transport des protéines , Facteurs de transcription/métabolisme , Ubiquitin-protein ligases/génétique , Ubiquitines/métabolisme , Danio zébré/génétique , Danio zébré/métabolisme
5.
Clin Genet ; 102(5): 444-450, 2022 11.
Article de Anglais | MEDLINE | ID: mdl-35908151

RÉSUMÉ

HIDEA syndrome is caused by biallelic pathogenic variants in P4HTM. The phenotype is characterized by muscular and central hypotonia, hypoventilation including obstructive and central sleep apneas, intellectual disability, dysautonomia, epilepsy, eye abnormalities, and an increased tendency to develop respiratory distress during pneumonia. Here, we report six new patients with HIDEA syndrome caused by five different biallelic P4HTM variants, including three novel variants. We describe two Finnish enriched pathogenic P4HTM variants and demonstrate that these variants are embedded within founder haplotypes. We review the clinical data from all previously published patients with HIDEA and characterize all reported P4HTM pathogenic variants associated with HIDEA in silico. All known pathogenic variants in P4HTM result in either premature stop codons, an intragenic deletion, or amino acid changes that impact the active site or the overall stability of P4H-TM protein. In all cases, normal P4H-TM enzyme function is expected to be lost or severely decreased. This report expands knowledge of the genotypic and phenotypic spectrum of the disease.


Sujet(s)
Codon non-sens , Déficience intellectuelle , Prolyl hydroxylases/métabolisme , Acides aminés , Domaine catalytique , Humains , Déficience intellectuelle/génétique , Déficience intellectuelle/anatomopathologie , Hypotonie musculaire/génétique , Phénotype , Syndrome
6.
Mol Cell ; 70(3): 462-472.e8, 2018 05 03.
Article de Anglais | MEDLINE | ID: mdl-29706539

RÉSUMÉ

Accumulation of the Xist long noncoding RNA (lncRNA) on one X chromosome is the trigger for X chromosome inactivation (XCI) in female mammals. Xist expression, which needs to be tightly controlled, involves a cis-acting region, the X-inactivation center (Xic), containing many lncRNA genes that evolved concomitantly to Xist from protein-coding ancestors through pseudogeneization and loss of coding potential. Here, we uncover an essential role for the Xic-linked noncoding gene Ftx in the regulation of Xist expression. We show that Ftx is required in cis to promote Xist transcriptional activation and establishment of XCI. Importantly, we demonstrate that this function depends on Ftx transcription and not on the RNA products. Our findings illustrate the multiplicity of layers operating in the establishment of XCI and highlight the diversity in the modus operandi of the noncoding players.


Sujet(s)
ARN long non codant/génétique , Inactivation du chromosome X/génétique , Chromosome X/génétique , Animaux , Lignée cellulaire , Femelle , Cellules HEK293 , Humains , Mammifères/génétique , Souris , Transcription génétique/génétique
7.
Stem Cells Int ; 2016: 9451492, 2016.
Article de Anglais | MEDLINE | ID: mdl-26798367

RÉSUMÉ

Pluripotent stem cells have the ability to undergo self-renewal and to give rise to all cells of the tissues of the body. However, this definition has been recently complicated by the existence of distinct cellular states that display these features. Here, we provide a detailed overview of the family of pluripotent cell lines derived from early mouse and human embryos and compare them with induced pluripotent stem cells. Shared and distinct features of these cells are reported as additional hallmark of pluripotency, offering a comprehensive scenario of pluripotent stem cells.

8.
Stem Cell Res ; 12(2): 323-37, 2014 Mar.
Article de Anglais | MEDLINE | ID: mdl-24365598

RÉSUMÉ

Understanding the molecular basis of cardiomyocyte development is critical for understanding the pathogenesis of pre- and post-natal cardiac disease. MicroRNAs (miRNAs) are post-transcriptional modulators of gene expression that play an important role in many developmental processes. Here, we show that the miR-99a/let-7c cluster, mapping on human chromosome 21, is involved in the control of cardiomyogenesis by altering epigenetic factors. By perturbing miRNA expression in mouse embryonic stem cells, we find that let-7c promotes cardiomyogenesis by upregulating genes involved in mesoderm specification (T/Bra and Nodal) and cardiac differentiation (Mesp1, Nkx2.5 and Tbx5). The action of let-7c is restricted to the early phase of mesoderm formation at the expense of endoderm and its late activation redirects cells toward other mesodermal derivatives. The Polycomb complex group protein Ezh2 is a direct target of let-7c, which promotes cardiac differentiation by modifying the H3K27me3 marks from the promoters of crucial cardiac transcription factors (Nkx2.5, Mef2c, Tbx5). In contrast, miR-99a represses cardiac differentiation via the nucleosome-remodeling factor Smarca5, attenuating the Nodal/Smad2 signaling. We demonstrated that the identified targets are underexpressed in human Down syndrome fetal heart specimens. By perturbing the expression levels of these miRNAs in embryonic stem cells, we were able to demonstrate that these miRNAs control lineage- and stage-specific transcription factors, working in concert with chromatin modifiers to direct cardiomyogenesis.


Sujet(s)
Cellules souches embryonnaires/physiologie , microARN/génétique , Myocytes cardiaques/physiologie , Animaux , Différenciation cellulaire/génétique , Cellules souches embryonnaires/cytologie , Cellules souches embryonnaires/métabolisme , Épigenèse génétique , Humains , Souris , Myocytes cardiaques/cytologie , Myocytes cardiaques/métabolisme , Transduction du signal , Transfection
9.
Biochimie ; 93(11): 1935-42, 2011 Nov.
Article de Anglais | MEDLINE | ID: mdl-21820484

RÉSUMÉ

Random X chromosome inactivation (XCI), the eutherian mechanism of X-linked gene dosage compensation, is controlled by a cis-acting locus termed the X-inactivation center (Xic). One of the striking features that characterize the Xic landscape is the abundance of loci transcribing non-coding RNAs (ncRNAs), including Xist, the master regulator of the inactivation process. Recent comparative genomic analyses have depicted the evolutionary scenario behind the origin of the X-inactivation center, revealing that this locus evolved from a region harboring protein-coding genes. During mammalian radiation, this ancestral protein-coding region was disrupted in the marsupial group, whilst it provided in eutherian lineage the starting material for the non-translated RNAs of the X-inactivation center. The emergence of non-coding genes occurred by a dual mechanism involving loss of protein-coding function of the pre-existing genes and integration of different classes of mobile elements, some of which modeled the structure and sequence of the non-coding genes in a species-specific manner. The rising genes started to produce transcripts that acquired function in regulating the epigenetic status of the X chromosome, as shown for Xist, its antisense Tsix, Jpx, and recently suggested for Ftx. Thus, the appearance of the Xic, which occurred after the divergence between eutherians and marsupials, was the basis for the evolution of random X inactivation as a strategy to achieve dosage compensation.


Sujet(s)
Évolution moléculaire , Cadres ouverts de lecture/génétique , ARN non traduit/génétique , ARN non traduit/métabolisme , Inactivation du chromosome X/génétique , Animaux , Compensation de dosage génétique , Régulation de l'expression des gènes , Humains , Marsupialia/génétique , Souris , ARN long non codant
10.
Hum Mol Genet ; 20(4): 705-18, 2011 Feb 15.
Article de Anglais | MEDLINE | ID: mdl-21118898

RÉSUMÉ

X chromosome inactivation (XCI) is an essential epigenetic process which involves several non-coding RNAs (ncRNAs), including Xist, the master regulator of X-inactivation initiation. Xist is flanked in its 5' region by a large heterochromatic hotspot, which contains several transcription units including a gene of unknown function, Ftx (five prime to Xist). In this article, we describe the characterization and functional analysis of murine Ftx. We present evidence that Ftx produces a conserved functional long ncRNA, and additionally hosts microRNAs (miR) in its introns. Strikingly, Ftx partially escapes X-inactivation and is upregulated specifically in female ES cells at the onset of X-inactivation, an expression profile which closely follows that of Xist. We generated Ftx null ES cells to address the function of this gene. In these cells, only local changes in chromatin marks are detected within the hotspot, indicating that Ftx is not involved in the global maintenance of the heterochromatic structure of this region. The Ftx mutation, however, results in widespread alteration of transcript levels within the X-inactivation center (Xic) and particularly important decreases in Xist RNA levels, which were correlated with increased DNA methylation at the Xist CpG island. Altogether our results indicate that Ftx is a positive regulator of Xist and lead us to propose that Ftx is a novel ncRNA involved in XCI.


Sujet(s)
Chromatine/composition chimique , ARN non traduit/génétique , ARN non traduit/métabolisme , Animaux , Bovins , Lignée cellulaire , Chromatine/métabolisme , Méthylation de l'ADN , Compensation de dosage génétique , Cellules souches embryonnaires/métabolisme , Femelle , Régulation de l'expression des gènes , Humains , Mâle , Souris , Mutation , Régions promotrices (génétique) , ARN long non codant , Similitude de séquences , Régulation positive , Inactivation du chromosome X/génétique
11.
Genome Biol ; 11(6): R64, 2010.
Article de Anglais | MEDLINE | ID: mdl-20569505

RÉSUMÉ

BACKGROUND: Dosage imbalance is responsible for several genetic diseases, among which Down syndrome is caused by the trisomy of human chromosome 21. RESULTS: To elucidate the extent to which the dosage imbalance of specific human chromosome 21 genes perturb distinct molecular pathways, we developed the first mouse embryonic stem (ES) cell bank of human chromosome 21 genes. The human chromosome 21-mouse ES cell bank includes, in triplicate clones, 32 human chromosome 21 genes, which can be overexpressed in an inducible manner. Each clone was transcriptionally profiled in inducing versus non-inducing conditions. Analysis of the transcriptional response yielded results that were consistent with the perturbed gene's known function. Comparison between mouse ES cells containing the whole human chromosome 21 (trisomic mouse ES cells) and mouse ES cells overexpressing single human chromosome 21 genes allowed us to evaluate the contribution of single genes to the trisomic mouse ES cell transcriptome. In addition, for the clones overexpressing the Runx1 gene, we compared the transcriptome changes with the corresponding protein changes by mass spectroscopy analysis. CONCLUSIONS: We determined that only a subset of genes produces a strong transcriptional response when overexpressed in mouse ES cells and that this effect can be predicted taking into account the basal gene expression level and the protein secondary structure. We showed that the human chromosome 21-mouse ES cell bank is an important resource, which may be instrumental towards a better understanding of Down syndrome and other human aneuploidy disorders.


Sujet(s)
Chromosomes humains de la paire 21/génétique , Cellules souches embryonnaires/cytologie , Cellules souches embryonnaires/métabolisme , Régulation de l'expression des gènes , Banques de tissus , Animaux , Lignée cellulaire , Sous-unité alpha 2 du facteur CBF/génétique , Sous-unité alpha 2 du facteur CBF/métabolisme , Dosage génique/génétique , Analyse de profil d'expression de gènes , Humains , Souris , Souris transgéniques , Réaction de polymérisation en chaîne , Protéome/métabolisme , Recombinaison génétique , Reproductibilité des résultats , Facteurs temps , Transcription génétique
12.
PLoS One ; 5(2): e9029, 2010 Feb 03.
Article de Anglais | MEDLINE | ID: mdl-20140255

RÉSUMÉ

Sik1 (salt inducible kinase 1) is a serine/threonine kinase that belongs to the stress- and energy-sensing AMP-activated protein kinase family. During murine embryogenesis, sik1 marks the monolayer of future myocardial cells that will populate first the primitive ventricle, and later the primitive atrium suggesting its involvement in cardiac cell differentiation and/or heart development. Despite that observation, the involvement of sik1 in cardiac differentiation is still unknown. We examined the sik1 function during cardiomyocyte differentiation using the ES-derived embryoid bodies. We produced a null embryonic stem cell using a gene-trap cell line carrying an insertion in the sik1 locus. In absence of the sik1 protein, the temporal appearance of cardiomyocytes is delayed. Expression profile analysis revealed sik1 as part of a genetic network that controls the cell cycle, where the cyclin-dependent kinase inhibitor p57(Kip2) is directly involved. Collectively, we provided evidence that sik1-mediated effects are specific for cardiomyogenesis regulating cardiomyoblast cell cycle exit toward terminal differentiation.


Sujet(s)
Inhibiteur p57 de kinase cycline-dépendante/métabolisme , Cellules souches embryonnaires/métabolisme , Myocytes cardiaques/métabolisme , Protein-Serine-Threonine Kinases/métabolisme , Animaux , Technique de Western , Cycle cellulaire , Différenciation cellulaire , Lignée cellulaire , Prolifération cellulaire , Cellules cultivées , Inhibiteur p57 de kinase cycline-dépendante/génétique , Régulation négative , Cellules souches embryonnaires/cytologie , Cytométrie en flux , Analyse de profil d'expression de gènes , Humains , Souris , Souris knockout , Myocytes cardiaques/cytologie , Neurones/cytologie , Neurones/métabolisme , Séquençage par oligonucléotides en batterie , Protein-Serine-Threonine Kinases/génétique , RT-PCR , Facteurs temps
13.
Nucleic Acids Res ; 33(4): e44, 2005 Mar 01.
Article de Anglais | MEDLINE | ID: mdl-15741177

RÉSUMÉ

In an effort to make transgenesis more flexible and reproducible, we developed a system based on novel 5' and 3' 'gene trap' vectors containing heterospecific Flp recognition target sites and the corresponding 'exchange' vectors allowing the insertion of any DNA sequence of interest into the trapped locus. Flp-recombinase-mediated cassette exchange was demonstrated to be highly efficient in our system, even in the absence of locus-specific selection. The feasibility of constructing a library of ES cell clones using our gene trap vectors was tested and a thousand insertion sites were characterized, following electroporation in ES cells, by RACE-PCR and sequencing. We validated the system in vivo for two trapped loci in transgenic mice and demonstrated that the reporter transgenes inserted into the trapped loci have an expression pattern identical to the endogenous genes. We believe that this system will facilitate in vivo studies of gene function and large-scale generation of mouse models of human diseases, caused by not only loss but also gain of function alleles.


Sujet(s)
Ciblage de gène/méthodes , Vecteurs génétiques , Souris transgéniques , Animaux , Lignée cellulaire , DNA nucleotidyltransferases/métabolisme , Souris , Recombinaison génétique
14.
Trends Genet ; 21(1): 12-6, 2005 Jan.
Article de Anglais | MEDLINE | ID: mdl-15680507

RÉSUMÉ

In metazoan organisms, energy production is the only example of a process that is under dual genetic control: nuclear and mitochondrial. We used a genomic approach to examine how energy genes of both the nuclear and mitochondrial genomes are coordinated, and discovered a novel genetic regulatory circuit in Drosophila melanogaster that is surprisingly simple and parsimonious. This circuit is based on a single DNA regulatory element and can explain both intra- and inter-genomic coordinated expression of genes involved in energy production, including the full complement of mitochondrial and nuclear oxidative phosphorylation genes, and the genes involved in the Krebs cycle.


Sujet(s)
Drosophila melanogaster/génétique , Métabolisme énergétique/génétique , Génome , Animaux , Anopheles/génétique , ADN mitochondrial/génétique , Drosophila/génétique
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