RESUMEN
Cilia biogenesis is a complex, multistep process involving the coordination of multiple cellular trafficking pathways. Despite the importance of ciliogenesis in mediating the cellular response to cues from the microenvironment, we have only a limited understanding of the regulation of cilium assembly. We previously identified Tau tubulin kinase 2 (TTBK2) as a key regulator of ciliogenesis. Here, using CRISPR kinome and biotin identification screening, we identify the CK2 catalytic subunit CSNK2A1 as an important modulator of TTBK2 function in cilia trafficking. Superresolution microscopy reveals that CSNK2A1 is a centrosomal protein concentrated at the mother centriole and associated with the distal appendages. Csnk2a1 mutant cilia are longer than those of control cells, showing instability at the tip associated with ciliary actin cytoskeleton changes. These cilia also abnormally accumulate key cilia assembly and SHH-related proteins. De novo mutations of Csnk2a1 were recently linked to the human genetic disorder Okur-Chung neurodevelopmental syndrome (OCNDS). Consistent with the role of CSNK2A1 in cilium stability, we find that expression of OCNDS-associated Csnk2a1 variants in wild-type cells causes ciliary structural defects. Our findings provide insights into mechanisms involved in ciliary length regulation, trafficking, and stability that in turn shed light on the significance of cilia instability in human disease.
Asunto(s)
Cilios/metabolismo , Ciliopatías/fisiopatología , Animales , Quinasa de la Caseína II/metabolismo , Quinasa de la Caseína II/fisiología , Línea Celular , Centriolos/metabolismo , Cilios/fisiología , Células HEK293 , Proteínas Hedgehog/metabolismo , Humanos , Ratones , Trastornos del Neurodesarrollo/genética , Trastornos del Neurodesarrollo/fisiopatología , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas/fisiologíaRESUMEN
Primary cilia contain specific proteins to achieve their functions as cellular antennae. Ciliary protein trafficking is mediated by the intraflagellar transport (IFT) machinery containing the IFT-A and IFT-B complexes. Mutations in genes encoding the IFT-A subunits (IFT43, IFT121/WDR35, IFT122, IFT139/TTC21B, IFT140 and IFT144/WDR19) often result in skeletal ciliopathies, including cranioectodermal dysplasia (CED). We here characterized the molecular and cellular defects of CED caused by compound heterozygous mutations in IFT144 [the missense variant IFT144(L710S) and the nonsense variant IFT144(R1103*)]. These two variants were distinct with regard to their interactions with other IFT-A subunits and with the IFT-B complex. When exogenously expressed in IFT144-knockout (KO) cells, IFT144(L710S) as well as IFT144(WT) rescued both moderately compromised ciliogenesis and the abnormal localization of ciliary proteins. As the homozygous IFT144(L710S) mutation was found to cause autosomal recessive retinitis pigmentosa, IFT144(L710S) is likely to be hypomorphic at the cellular level. In striking contrast, the exogenous expression of IFT144(R1103*) in IFT144-KO cells exacerbated the ciliogenesis defects. The expression of IFT144(R1103*) together with IFT144(WT) restored the abnormal phenotypes of IFT144-KO cells. However, the coexpression of IFT144(R1103*) with the hypomorphic IFT144(L710S) variant in IFT144-KO cells, which mimics the genotype of compound heterozygous CED patients, resulted in severe ciliogenesis defects. Taken together, these observations demonstrate that compound heterozygous mutations in IFT144 cause severe ciliary defects via a complicated mechanism, where one allele can cause severe ciliary defects when combined with a hypomorphic allele.
Asunto(s)
Huesos/anomalías , Cilios/metabolismo , Craneosinostosis/metabolismo , Proteínas del Citoesqueleto/genética , Proteínas del Citoesqueleto/metabolismo , Displasia Ectodérmica/metabolismo , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Mutación , Huesos/metabolismo , Huesos/fisiopatología , Cilios/patología , Ciliopatías/genética , Ciliopatías/metabolismo , Ciliopatías/fisiopatología , Codón sin Sentido , Craneosinostosis/genética , Craneosinostosis/fisiopatología , Displasia Ectodérmica/genética , Displasia Ectodérmica/fisiopatología , Células HEK293 , Humanos , Mutación MissenseRESUMEN
Mutations in subunits of the cilia-specific cytoplasmic dynein-2 (CD2) complex cause short-rib thoracic dystrophy syndromes (SRTDs), characterized by impaired bone growth and life-threatening perinatal respiratory complications. Different SRTD mutations result in varying disease severities. It remains unresolved whether this reflects the extent of retained hypomorphic protein functions or relative importance of the affected subunits for the activity of the CD2 holoenzyme. To define the contribution of the LC8-type dynein light chain subunit to the CD2 complex, we have generated Dynll1-deficient mouse strains, including the first-ever conditional knockout (KO) mutant for any CD2 subunit. Germline Dynll1 KO mice exhibit a severe ciliopathy-like phenotype similar to mice lacking another CD2 subunit, Dync2li1. Limb mesoderm-specific loss of Dynll1 results in severe bone shortening similar to human SRTD patients. Mechanistically, loss of Dynll1 leads to a partial depletion of other SRTD-related CD2 subunits, severely impaired retrograde intra-flagellar transport, significant thickening of primary cilia and cilia signaling defects. Interestingly, phenotypes of Dynll1-deficient mice are very similar to entirely cilia-deficient Kif3a/Ift88-null mice, except that they never present with polydactyly and retain relatively higher signaling outputs in parts of the hedgehog pathway. Compared to complete loss of Dynll1, maintaining very low DYNLL1 levels in mice lacking the Dynll1-transcription factor ASCIZ (ATMIN) results in significantly attenuated phenotypes and improved CD2 protein levels. The results suggest that primary cilia can maintain some functionality in the absence of intact CD2 complexes and provide a viable animal model for the analysis of the underlying bone development defects of SRTDs.
Asunto(s)
Enfermedades del Desarrollo Óseo/metabolismo , Cilios/metabolismo , Ciliopatías/metabolismo , Dineínas Citoplasmáticas/genética , Osteogénesis , Animales , Enfermedades del Desarrollo Óseo/genética , Enfermedades del Desarrollo Óseo/fisiopatología , Células Cultivadas , Cilios/fisiología , Ciliopatías/genética , Ciliopatías/fisiopatología , Dineínas Citoplasmáticas/metabolismo , Dineínas Citoplasmáticas/fisiología , Extremidades/patología , Extremidades/fisiopatología , Proteínas Hedgehog/metabolismo , Masculino , Ratones , Ratones Noqueados , Fenotipo , Transducción de Señal , Factores de Transcripción/metabolismoRESUMEN
Primary cilia are specialized sensory organelles that protrude from the apical surface of most cell types. During the past 2 decades, they have been found to play important roles in tissue development and signal transduction, with mutations in ciliary-associated proteins resulting in a group of diseases collectively known as ciliopathies. Many of these mutations manifest as renal ciliopathies, characterized by kidney dysfunction resulting from aberrant cilia or ciliary functions. This group of overlapping and genetically heterogeneous diseases includes polycystic kidney disease, nephronophthisis, and Bardet-Biedl syndrome as the main focus of this review. Renal ciliopathies are characterized by the presence of kidney cysts that develop due to uncontrolled epithelial cell proliferation, growth, and polarity, downstream of dysregulated ciliary-dependent signaling. Due to cystic-associated kidney injury and systemic inflammation, cases result in kidney failure requiring dialysis and transplantation. Of the handful of pharmacologic treatments available, none are curative. It is important to determine the molecular mechanisms that underlie the involvement of the primary cilium in cyst initiation, expansion, and progression for the development of novel and efficacious treatments. This review updates research progress in defining key genes and molecules central to ciliogenesis and renal ciliopathies.
Asunto(s)
Síndrome de Bardet-Biedl/genética , Cilios/metabolismo , Ciliopatías/genética , Enfermedades Renales Poliquísticas/genética , Anomalías Múltiples/genética , Anomalías Múltiples/metabolismo , Anomalías Múltiples/fisiopatología , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras del Transporte Vesicular/genética , Síndrome de Bardet-Biedl/metabolismo , Síndrome de Bardet-Biedl/fisiopatología , Cerebelo/anomalías , Cerebelo/metabolismo , Cerebelo/fisiopatología , Chaperoninas/genética , Cilios/fisiología , Trastornos de la Motilidad Ciliar/genética , Trastornos de la Motilidad Ciliar/metabolismo , Trastornos de la Motilidad Ciliar/fisiopatología , Ciliopatías/metabolismo , Ciliopatías/fisiopatología , Proteínas del Citoesqueleto/genética , Encefalocele/genética , Encefalocele/metabolismo , Encefalocele/fisiopatología , Anomalías del Ojo/genética , Anomalías del Ojo/metabolismo , Anomalías del Ojo/fisiopatología , Humanos , Enfermedades Renales Quísticas/genética , Enfermedades Renales Quísticas/metabolismo , Enfermedades Renales Quísticas/fisiopatología , Amaurosis Congénita de Leber/genética , Amaurosis Congénita de Leber/metabolismo , Amaurosis Congénita de Leber/fisiopatología , Proteínas de la Membrana/genética , Proteínas Asociadas a Microtúbulos/genética , Atrofias Ópticas Hereditarias/genética , Atrofias Ópticas Hereditarias/metabolismo , Atrofias Ópticas Hereditarias/fisiopatología , Enfermedades Renales Poliquísticas/metabolismo , Enfermedades Renales Poliquísticas/fisiopatología , Proteínas/genética , Retina/anomalías , Retina/metabolismo , Retina/fisiopatología , Retinitis Pigmentosa/genética , Retinitis Pigmentosa/metabolismo , Retinitis Pigmentosa/fisiopatología , Canales Catiónicos TRPP/genéticaRESUMEN
PURPOSE: To report genetic and clinical findings in a case series of 10 patients from eight unrelated families diagnosed with Senior-Løken syndrome. METHODS: A retrospective study of patients with Senior-Løken syndrome. Data collected included clinical findings electroretinography and ocular imaging. Genetic analysis was based on molecular inversion probes, whole-exome sequencing (WES), and Sanger sequencing. RESULTS: All patients who underwent electrophysiology (8/10) had widespread photoreceptor degeneration. Genetic analysis revealed two mutations in NPHP1, two mutations in NPHP4, and two mutations in IQCB1 (NPHP5). Five of the six mutations identified in the current study were found in a single family each in our cohort. The IQCB1-p.R461* mutation has been identified in 3 families. Patients harboring mutations in IQCB1 were diagnosed with Leber congenital amaurosis, while patients with NPHP4 and NPHP1 mutations showed early and sector retinitis pigmentosa, respectively. Full-field electroretinography was extinct for 6 of 10 patients, moderately decreased for two, and unavailable for another 2 subjects. Renal involvement was evident in 7/10 patients at the time of diagnosis. Kidney function was normal (based on serum creatinine) in patients younger than 10 years. Mutations in IQCB1 were associated with high hypermetropia, whereas mutations in NPHP4 were associated with high myopia. CONCLUSION: Patients presenting with infantile inherited retinal degeneration are not universally screened for renal dysfunction. Modern genetic tests can provide molecular diagnosis at an early age and therefore facilitate early diagnosis of renal disease with recommended periodic screening beyond childhood and family planning.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas de Unión a Calmodulina/genética , Ciliopatías/genética , Proteínas del Citoesqueleto/genética , Enfermedades Renales Quísticas/genética , Amaurosis Congénita de Leber/genética , Mutación , Atrofias Ópticas Hereditarias/genética , Proteínas/genética , Adolescente , Niño , Preescolar , Ciliopatías/diagnóstico , Ciliopatías/fisiopatología , Pruebas de Percepción de Colores , Análisis Mutacional de ADN , Electrorretinografía , Femenino , Humanos , Lactante , Enfermedades Renales Quísticas/diagnóstico , Enfermedades Renales Quísticas/fisiopatología , Amaurosis Congénita de Leber/diagnóstico , Amaurosis Congénita de Leber/fisiopatología , Masculino , Persona de Mediana Edad , Técnicas de Diagnóstico Molecular , Atrofias Ópticas Hereditarias/diagnóstico , Atrofias Ópticas Hereditarias/fisiopatología , Linaje , Fenotipo , Retina/fisiopatología , Estudios Retrospectivos , Agudeza Visual/fisiología , Pruebas del Campo Visual , Secuenciación del Exoma , Adulto JovenRESUMEN
Spinocerebellar ataxia type 11 (SCA11) is a rare, dominantly inherited human ataxia characterized by atrophy of Purkinje neurons in the cerebellum. SCA11 is caused by mutations in the gene encoding the Serine/Threonine kinase Tau tubulin kinase 2 (TTBK2) that result in premature truncations of the protein. We previously showed that TTBK2 is a key regulator of the assembly of primary cilia in vivo. However, the mechanisms by which the SCA11-associated mutations disrupt TTBK2 function, and whether they interfere with ciliogenesis were unknown. In this work, we present evidence that SCA11-associated mutations are dominant negative alleles and that the resulting truncated protein (TTBK2SCA11) interferes with the function of full length TTBK2 in mediating ciliogenesis. A Ttbk2 allelic series revealed that upon partial reduction of full length TTBK2 function, TTBK2SCA11 can interfere with the activity of the residual wild-type protein to decrease cilia number and interrupt cilia-dependent Sonic hedgehog (SHH) signaling. Our studies have also revealed new functions for TTBK2 after cilia initiation in the control of cilia length, trafficking of a subset of SHH pathway components, including Smoothened (SMO), and cilia stability. These studies provide a molecular foundation to understand the cellular and molecular pathogenesis of human SCA11, and help account for the link between ciliary dysfunction and neurodegenerative diseases.
Asunto(s)
Cilios/patología , Cilios/fisiología , Ciliopatías/genética , Ciliopatías/fisiopatología , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/fisiología , Ataxias Espinocerebelosas/genética , Ataxias Espinocerebelosas/fisiopatología , Alelos , Animales , Ciliopatías/patología , Modelos Animales de Enfermedad , Femenino , Técnicas de Sustitución del Gen , Genes Dominantes , Homocigoto , Humanos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Microscopía Electrónica de Transmisión , Mutación , Ataxias Espinocerebelosas/patologíaRESUMEN
BACKGROUND: Joubert syndrome and related disorders (JSRD) and Jeune syndrome are multisystem ciliopathy disorders with overlapping phenotypes. There are a growing number of genetic causes for these rare syndromes, including the recently described genes ARL3 and CEP120. METHODS: We sought to explore the developmental expression patterns of ARL3 and CEP120 in humans to gain additional understanding of these genetic conditions. We used an RNA in situ detection technique called RNAscope to characterise ARL3 and CEP120 expression patterns in human embryos and foetuses in collaboration with the MRC-Wellcome Trust Human Developmental Biology Resource. RESULTS: Both ARL3 and CEP120 are expressed in early human brain development, including the cerebellum and in the developing retina and kidney, consistent with the clinical phenotypes seen with pathogenic variants in these genes. CONCLUSIONS: This study provides insights into the potential pathogenesis of JSRD by uncovering the spatial expression of two JSRD-causative genes during normal human development.
Asunto(s)
Factores de Ribosilacion-ADP/genética , Proteínas de Ciclo Celular/genética , Ciliopatías/genética , Regulación del Desarrollo de la Expresión Génica , Factores de Ribosilacion-ADP/metabolismo , Encéfalo/crecimiento & desarrollo , Encéfalo/metabolismo , Proteínas de Ciclo Celular/metabolismo , Ciliopatías/patología , Ciliopatías/fisiopatología , Ganglios Espinales/crecimiento & desarrollo , Ganglios Espinales/metabolismo , Humanos , Riñón/crecimiento & desarrollo , Riñón/metabolismo , Mutación , Fenotipo , Retina/crecimiento & desarrollo , Retina/metabolismoRESUMEN
Skeletal dysplasias are a clinically and genetically heterogeneous group of bone and cartilage disorders. A total of 436 skeletal dysplasias are listed in the 2015 revised version of the nosology and classification of genetic skeletal disorders, of which nearly 20% are still genetically and molecularly uncharacterized. We report the clinical and molecular characterization of a lethal skeletal dysplasia of the short-rib group caused by mutation of the mouse Fop gene. Fop encodes a centrosomal and centriolar satellite (CS) protein. We show that Fop mutation perturbs ciliogenesis in vivo and that this leads to the alteration of the Hedgehog signaling pathway. Fop mutation reduces CSs movements and affects pericentriolar material composition, which probably participates to the ciliogenesis defect. This study highlights the role of a centrosome and CSs protein producing phenotypes in mice that recapitulate a short rib-polydactyly syndrome when mutated.
Asunto(s)
Ciliopatías/genética , Proteínas Proto-Oncogénicas/genética , Síndrome de Costilla Pequeña y Polidactilia/genética , Factores de Transcripción/genética , Animales , Centriolos/genética , Centrosoma/metabolismo , Centrosoma/patología , Cilios/genética , Cilios/patología , Ciliopatías/fisiopatología , Humanos , Ratones , Mutación , Síndrome de Costilla Pequeña y Polidactilia/fisiopatologíaRESUMEN
Cilia project from almost every cell integrating extracellular cues with signaling pathways. Constitutive activation of FGFR3 signaling produces the skeletal disorders achondroplasia (ACH) and thanatophoric dysplasia (TD), but many of the molecular mechanisms underlying these phenotypes remain unresolved. Here, we report in vivo evidence for significantly shortened primary cilia in ACH and TD cartilage growth plates. Using in vivo and in vitro methodologies, our data demonstrate that transient versus sustained activation of FGF signaling correlated with different cilia consequences. Transient FGF pathway activation elongated cilia, while sustained activity shortened cilia. FGF signaling extended primary cilia via ERK MAP kinase and mTORC2 signaling, but not through mTORC1. Employing a GFP-tagged IFT20 construct to measure intraflagellar (IFT) speed in cilia, we showed that FGF signaling affected IFT velocities, as well as modulating cilia-based Hedgehog signaling. Our data integrate primary cilia into canonical FGF signal transduction and uncover a FGF-cilia pathway that needs consideration when elucidating the mechanisms of physiological and pathological FGFR function, or in the development of FGFR therapeutics.
Asunto(s)
Acondroplasia/fisiopatología , Receptor Tipo 3 de Factor de Crecimiento de Fibroblastos/metabolismo , Displasia Tanatofórica/fisiopatología , Acondroplasia/genética , Animales , Cartílago/metabolismo , Condrocitos/metabolismo , Cilios/patología , Cilios/fisiología , Ciliopatías/genética , Ciliopatías/fisiopatología , Factores de Crecimiento de Fibroblastos/metabolismo , Placa de Crecimiento/metabolismo , Humanos , Ratones , Células 3T3 NIH , Fenotipo , Cultivo Primario de Células , Receptor Tipo 3 de Factor de Crecimiento de Fibroblastos/genética , Transducción de Señal/fisiología , Displasia Tanatofórica/genéticaRESUMEN
Short-rib polydactyly syndromes are a heterogeneous group of disorders characterized by narrow thorax with short ribs, polydactyly and often other visceral and skeletal malformations. To date there have only been six reported patients with homozygous and compound heterozygous variants in IFT81, causing a short-rib thoracic dysplasia, with, or without, polydactyly (SRTD19: OMIM 617895). IFT81 is a protein integral to the core of the intraflagellar transport complex B (IFT-B), which is involved in anterograde transport in the cilium. We describe the case of a male infant with compound heterozygous variants in IFT81, who presented with short long bones, a narrow thorax, polydactyly, and multiple malformations. Three novel clinical features are reported including complete situs inversus, micropenis, and rectal atresia, which have not previously been associated with variants in IFT81. We reviewed the literature and identified the most consistent clinical features associated with this rare ciliopathy syndrome. We postulate that dolichocephaly and sagittal craniosynostosis may be associated with this condition, and provide a clue to considering IFT81 as the causative gene when deciphering complex ciliopathies.
Asunto(s)
Ciliopatías/genética , Craneosinostosis/genética , Proteínas Musculares/genética , Síndrome de Costilla Pequeña y Polidactilia/genética , Cilios/patología , Ciliopatías/diagnóstico , Ciliopatías/fisiopatología , Craneosinostosis/diagnóstico , Craneosinostosis/fisiopatología , Homocigoto , Humanos , Recién Nacido , Masculino , Mutación/genética , Fenotipo , Síndrome de Costilla Pequeña y Polidactilia/diagnóstico , Síndrome de Costilla Pequeña y Polidactilia/fisiopatologíaRESUMEN
Nephronophthisis (NPH) is an autosomal recessive renal disease leading to kidney failure in children and young adults. The protein products of the corresponding genes (NPHPs) are localized in primary cilia or their appendages. Only about 70% of affected individuals have a mutation in one of 100 renal ciliopathy genes, and no unifying pathogenic mechanism has been identified. Recently, some NPHPs, including NIMA-related kinase 8 (NEK8) and centrosomal protein 164 (CEP164), have been found to act in the DNA-damage response pathway and to contribute to genome stability. Here, we show that NME/NM23 nucleoside-diphosphate kinase 3 (NME3) that has recently been found to facilitate DNA-repair mechanisms binds to several NPHPs, including NEK8, CEP164, and ankyrin repeat and sterile α motif domain-containing 6 (ANKS6). Depletion of nme3 in zebrafish and Xenopus resulted in typical ciliopathy-associated phenotypes, such as renal malformations and left-right asymmetry defects. We further found that endogenous NME3 localizes to the basal body and that it associates also with centrosomal proteins, such as NEK6, which regulates cell cycle arrest after DNA damage. The ciliopathy-typical manifestations of NME3 depletion in two vertebrate in vivo models, the biochemical association of NME3 with validated NPHPs, and its localization to the basal body reveal a role for NME3 in ciliary function. We conclude that mutations in the NME3 gene may aggravate the ciliopathy phenotypes observed in humans.
Asunto(s)
Ciliopatías/genética , Enfermedades Renales Quísticas/congénito , Nucleósido Difosfato Quinasas NM23/genética , Insuficiencia Renal/genética , Animales , Puntos de Control del Ciclo Celular/genética , Cilios/genética , Cilios/patología , Ciliopatías/fisiopatología , Daño del ADN/genética , Reparación del ADN/genética , Modelos Animales de Enfermedad , Humanos , Riñón/patología , Enfermedades Renales Quísticas/genética , Enfermedades Renales Quísticas/patología , Proteínas de Microtúbulos/genética , Quinasas Relacionadas con NIMA/genética , Proteínas Nucleares/genética , Insuficiencia Renal/patología , Xenopus/genética , Pez Cebra/genéticaAsunto(s)
Anomalías Múltiples , Cerebelo , Ciliopatías , Anomalías del Ojo , Hidrocefalia , Enfermedades Renales Quísticas , Fenotipo , Retina , Humanos , Hidrocefalia/genética , Hidrocefalia/diagnóstico por imagen , Hidrocefalia/fisiopatología , Enfermedades Renales Quísticas/genética , Enfermedades Renales Quísticas/complicaciones , Anomalías del Ojo/genética , Cerebelo/anomalías , Cerebelo/fisiopatología , Cerebelo/diagnóstico por imagen , Anomalías Múltiples/genética , Retina/fisiopatología , Retina/anomalías , Ciliopatías/genética , Ciliopatías/fisiopatología , Masculino , Imagen por Resonancia Magnética , FemeninoRESUMEN
The short-rib polydactyly syndromes (SRPS) encompass a radiographically and genetically heterogeneous group of skeletal ciliopathies that are characterized by a long narrow chest, short extremities, and variable occurrence of polydactyly. Radiographic abnormalities include undermineralization of the calvarium, shortened and bowed appendicular bones, trident shaped acetabula and polydactyly. In a case of SRPS we identified compound heterozygosity for mutations in IFT52, which encodes a component of the anterograde intraflagellar transport complex. The IFT52 mutant cells synthesized a significantly reduced amount of IFT52 protein, leading to reduced synthesis of IFT74, IFT81, IFT88 and ARL13B, other key anterograde complex members. Ciliogenesis was also disrupted in the mutant cells, with a 60% reduction in the presence of cilia on mutant cells and loss of cilia length regulation for the cells with cilia. These data demonstrate that IFT52 is essential for anterograde complex integrity and for the biosynthesis and maintenance of cilia. The data identify a new locus for SRPS and show that IFT52 mutations result in a ciliopathy with primary effects on the skeleton.
Asunto(s)
Proteínas Portadoras/genética , Cilios/genética , Ciliopatías/genética , Síndrome de Costilla Pequeña y Polidactilia/genética , Cilios/metabolismo , Ciliopatías/fisiopatología , Proteínas del Citoesqueleto/genética , Flagelos/genética , Flagelos/patología , Humanos , Péptidos y Proteínas de Señalización Intracelular , Complejos Multiproteicos/genética , Proteínas Musculares/genética , Mutación/genética , Síndrome de Costilla Pequeña y Polidactilia/fisiopatología , Esqueleto/crecimiento & desarrollo , Esqueleto/metabolismo , Esqueleto/patología , Proteínas Supresoras de Tumor/genéticaRESUMEN
Whole genome sequencing (WGS) was performed to identify the variants responsible for inherited retinal degeneration (IRD) in a Caucasian family. Segregation analysis of selected rare variants with pathogenic potential identified a set of compound heterozygous changes p.Arg266*:c.796C>T and p.Ala568Thr:c.1702G>A in the intraflagellar transport protein-88 (IFT88) gene segregating with IRD. Expression of IFT88 with the p.Arg266* and p.Ala568Thr mutations in mIMDC3 cells by transient transfection and in HeLa cells by introducing the mutations using CRISPR-cas9 system suggested that both mutations result in the formation of abnormal ciliary structures. The introduction of the IFT88 p.Arg266* variant in the homozygous state in HeLa cells by CRISPR-Cas9 genome-editing revealed that the mutant transcript undergoes nonsense-mediated decay leading to a significant depletion of IFT88 transcript. Additionally, abnormal ciliogenesis was observed in these cells. These observations suggest that the rare and unique combination of IFT88 alleles observed in this study provide insight into the physiological role of IFT88 in humans and the likely mechanism underlying retinal pathology in the pedigree with IRD.
Asunto(s)
Ciliopatías/genética , Degeneración Retiniana/genética , Proteínas Supresoras de Tumor/genética , Secuenciación Completa del Genoma , Alelos , Sistemas CRISPR-Cas/genética , Ciliopatías/fisiopatología , Femenino , Edición Génica , Predisposición Genética a la Enfermedad , Células HeLa , Homocigoto , Humanos , Masculino , Persona de Mediana Edad , Mutación , Linaje , Retina/patología , Degeneración Retiniana/fisiopatologíaRESUMEN
Ciliopathies are disorders of the primary cilium that can affect almost all organs and that are characterized by pleiotropy and extensive intra- and interfamilial phenotypic variability. Accordingly, mutations in the same gene can cause different ciliopathy phenotypes of varying severity. WDR60 encodes a protein thought to play a role in the primary cilium's intraflagellar transport machinery. Mutations in this gene are a rare cause of Jeune asphyxiating thoracic dystrophy (JATD) and short-rib polydactyly syndrome (SRPS). Here we report on a milder and distinct phenotype in a consanguineous Pakistani pedigree with two adolescent sisters affected by retinal degeneration and postaxial polydactyly, but lack of any further skeletal or chondrodysplasia features. By targeted high-throughput sequencing of genes known or suspected to be involved in ciliogenesis, we detected a novel homozygous N-terminal truncating WDR60 mutation (c.44delC/p.Ala15Glufs*90) that co-segregated with the disease in the family. Our finding broadens the spectrum of WDR60-related phenotypes and shows the utility of broad multigene panels during the genetic work-up of patients with ciliopathies.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Polidactilia/genética , Degeneración Retiniana/genética , Síndrome de Costilla Pequeña y Polidactilia/genética , Adolescente , Adulto , Cilios/genética , Cilios/patología , Ciliopatías/genética , Ciliopatías/fisiopatología , Síndrome de Ellis-Van Creveld/genética , Síndrome de Ellis-Van Creveld/fisiopatología , Exoma/genética , Femenino , Homocigoto , Humanos , Masculino , Persona de Mediana Edad , Mutación , Linaje , Polidactilia/fisiopatología , Degeneración Retiniana/fisiopatología , Costillas/fisiopatología , Síndrome de Costilla Pequeña y Polidactilia/fisiopatología , Hermanos , Adulto JovenRESUMEN
The function of normal and defective candidate genes for human genetic diseases, which are rapidly being identified in large numbers by human geneticists and the biomedical community at large, will be best studied in relevant and predictive model organisms that allow high-speed verification, analysis of underlying developmental, cellular and molecular mechanisms, and establishment of disease models to test therapeutic options. We describe and discuss the pros and cons of the frog Xenopus, which has been extensively used to uncover developmental mechanisms in the past, but which is being underutilized as a biomedical model. We argue that Xenopus complements the more commonly used mouse and zebrafish as a time- and cost-efficient animal model to study human disease alleles and mechanisms.
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Anomalías Congénitas/genética , Modelos Animales de Enfermedad , Enfermedades Genéticas Congénitas/genética , Xenopus laevis/genética , Alelos , Animales , Trastornos de la Motilidad Ciliar/embriología , Trastornos de la Motilidad Ciliar/genética , Trastornos de la Motilidad Ciliar/fisiopatología , Ciliopatías/embriología , Ciliopatías/genética , Ciliopatías/fisiopatología , Anomalías Congénitas/embriología , Anomalías Congénitas/fisiopatología , Enfermedades Genéticas Congénitas/embriología , Enfermedades Genéticas Congénitas/fisiopatología , Cardiopatías Congénitas/embriología , Cardiopatías Congénitas/genética , Cardiopatías Congénitas/fisiopatología , Humanos , Mutación , Xenopus laevis/embriología , Xenopus laevis/fisiologíaRESUMEN
Ciliopathies are a group of disorders caused by a defect in ciliogenesis, ciliary protein trafficking. Because nearly every cell in the body (including the photoreceptors) contains cilia, defects in ciliary proteins typically affect multiple organ systems. Usher syndrome is the most common syndromic cause of retinitis pigmentosa (RP) and accounts for 10-20% of cases of RP Inheritance is autosomal recessive, and the retinal dystrophy is usually rod-cone dystrophy (Figs. 32.1 and 32.2). These patients have RP with sensorineural hearing loss (partial or complete) since birth; some may have vestibular dysfunction. Most patients retain central vision of about 20/40 until about age 40. Usher Syndrome 1 (USH1): Profound congenital sensorineural hearing loss on audiometry, absent vestibular function, and typical RP (onset by 10 years of age); accounts for about 70% of all Usher cases. Patient may benefit from a cochlear implant. The retinitis pigmentosa occurs at an early age (childhood onset) and progress slowly. Usher Syndrome 2 (USH2): Moderate to severe congenital sensorineural hearing loss on audiometry (predominantly for higher frequencies), normal vestibular function, and typical RP (onset by 20 years of age); accounts for about 26% of all Usher cases. Usher Syndrome 3 (USH3): Progressive sensorineural hearing loss and typical RP (onset in second decade); accounts for about 4% of all Usher cases. Vestibular function is normal in about half of patients, but abnormal in the other half.
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Ciliopatías/fisiopatología , Síndromes de Usher/fisiopatología , Edad de Inicio , HumanosRESUMEN
Glistening yellow-white crystalline inclusions in foveal and parafoveal areas are almost pathognomonic (Fig. 36.1). These inclusions are evident at 1-2 years old and increase with age. Patients may have corneal stromal opacities, punctate keratitis, myopia, and astigmatism. About 50% have pigmentary degeneration of the retina, with decreased visual acuity and marked photophobia. Patients have dry, scaly skin (ichthyosis). Affected infants tend to be born prematurely. They also have neurological problems due to leukoencephalopathy (affecting the white matter of the brain). Intellectual disability varies from mild to severe, along with dysarthria and delayed speech.
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Ciliopatías/fisiopatología , Síndrome de Sjögren-Larsson/fisiopatología , Fóvea Central/patología , Humanos , Retina/patologíaRESUMEN
Senior-Løken syndrome is a rare autosomal recessive disease with a prevalence of 1:1,000,000. Retinopathy may progress as Leber congenital amaurosis (LCA), retinitis pigmentosa (RP), or sector RP (Figs. 34.1 and 34.2). Onset of photophobia, nystagmus, and hyperopia can occur in the first few years of life or later in childhood. Patients experience nephronophthisis, characterized by cystic kidney disease (medullary cystic kidney disease), reduced concentrating ability, and chronic tubulointerstitial nephritis, which progresses to end-stage renal disease. Hypertension is common.
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Ciliopatías/fisiopatología , Enfermedades Renales Quísticas/fisiopatología , Amaurosis Congénita de Leber/fisiopatología , Atrofias Ópticas Hereditarias/fisiopatología , Humanos , MutaciónRESUMEN
Ciliopathies are disorders caused by ciliary dysfunction and can affect an organ system or tissues. Isolated or syndromic retinal dystrophies are the most common ocular manifestation of ciliopathies. The photoreceptor connecting cilium plays a leading role in these ciliopathy-related retinal dystrophies. Dysfunctional photoreceptor cilia cause the most severe type of retinal dystrophy: Leber's congenital amaurosis (LCA). The most common syndromic ciliopathies with an ocular manifestation are Bardet-Biedl syndrome (BBS) and Usher syndrome. Molecular-genetic analysis revealed a large number of cilia genes with a high phenotype heterogeneity. Diagnosis of ciliopathies require a multi-disciplinary approach. Causative treatment of ciliopathies is not yet available; therefore, rehabilitative and supportive treatment is mandatory.