RESUMEN
Purpose: We comprehensively evaluated the mutational spectrum of Leber congenital amaurosis (LCA) and investigated the molecular diagnostic rate and genotype-phenotype correlation in a Korean cohort. Methods: This single-center retrospective case series included 50 Korean patients with LCA between June 2015 and March 2019. Molecular analysis was conducted using targeted panel-based next-generation sequencing, including deep intronic and regulatory variants or whole exome sequencing. The molecular diagnosis was made based on the inheritance pattern, zygosity, and pathogenicity. Results: Among the 50 patients, 27 patients (54%) were male, and 11 (22%) showed systemic features. Genetic variants highly likely to be causative were identified in 78% (39/50) of cases and segregated into families. We detected two pathogenic or likely pathogenic variants in a gene linked to a recessive trait without segregation analysis in three cases (6.0%). GUCY2D (20%), NMNAT1 (18%), and CEP290 (16%) were the most frequently mutated genes in Korean LCA. Copy number variations were found in three patients, which accounted for 6% of LCA cases. A possible dual molecular diagnosis (Senior-Løken syndrome along with Leigh syndrome, and Joubert syndrome with transposition of the great arteries) was made in two patients (4%). Three of 50 patients were medically or surgically actionable: one patient for RPE65 gene therapy and two patients with WDR19 Senior-Løken syndrome for early preparation for kidney and liver transplantations. Conclusions: This study demonstrated that approximately 4% of patients may have dual molecular diagnoses, and 6% were surgically or medically actionable in LCA. Therefore, accurate molecular diagnosis and careful interpretation of next-generation sequencing results can be of great help in patients with LCA.
Asunto(s)
Anomalías Múltiples/genética , Cerebelo/anomalías , Ciliopatías/genética , Variaciones en el Número de Copia de ADN/genética , Anomalías del Ojo/genética , Enfermedades Renales Quísticas/genética , Amaurosis Congénita de Leber/diagnóstico , Amaurosis Congénita de Leber/genética , Enfermedad de Leigh/genética , Atrofias Ópticas Hereditarias/genética , Retina/anomalías , Anomalías Múltiples/diagnóstico , Adolescente , Adulto , Antígenos de Neoplasias/sangre , Antígenos de Neoplasias/genética , Proteínas de Ciclo Celular/sangre , Proteínas de Ciclo Celular/genética , Niño , Preescolar , Ciliopatías/diagnóstico , Proteínas del Citoesqueleto/sangre , Proteínas del Citoesqueleto/genética , Anomalías del Ojo/diagnóstico , Femenino , Estudios de Asociación Genética , Terapia Genética , Guanilato Ciclasa/sangre , Guanilato Ciclasa/genética , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Lactante , Péptidos y Proteínas de Señalización Intracelular/genética , Enfermedades Renales Quísticas/diagnóstico , Amaurosis Congénita de Leber/diagnóstico por imagen , Amaurosis Congénita de Leber/terapia , Enfermedad de Leigh/diagnóstico , Masculino , Mutación , Nicotinamida-Nucleótido Adenililtransferasa/sangre , Nicotinamida-Nucleótido Adenililtransferasa/genética , Atrofias Ópticas Hereditarias/diagnóstico , Trasplante de Órganos , Linaje , Receptores de Superficie Celular/sangre , Receptores de Superficie Celular/genética , República de Corea , Estudios Retrospectivos , Transposición de los Grandes Vasos/genética , cis-trans-Isomerasas/genéticaRESUMEN
Sprint interval training (SIT) is reported to improve blood glucose control and may be a useful public health tool. The sirtuins and associated genes are emerging as key players in blood glucose control. This study investigated the interplay between the sirtuin/NAD system and individual variation in insulin sensitivity responses after SIT in young healthy individuals. Before and after 4 weeks of SIT, body mass and fat percentage were measured and oral glucose tolerance tests performed in 20 young healthy participants (7 females). Blood gene expression profiles (all 7 mammalian sirtuin genes and 15 enzymes involved in conversion of tryptophan, bioavailable vitamin B3, and metabolic precursors to NAD). NAD/NADP was measured in whole blood. Significant reductions in body weight and body fat post-SIT were associated with altered lipid profiles, NAD/NADP, and regulation of components of the sirtuin/NAD system (NAMPT, NMNAT1, CD38, and ABCA1). Variable improvements in measured metabolic health parameters were evident and attributed to different responses in males and females, together with marked inter-individual variation in responses of the sirtuin/NAD system to SIT.
Asunto(s)
Entrenamiento de Intervalos de Alta Intensidad/métodos , Carrera , Sirtuinas/sangre , ADP-Ribosil Ciclasa 1/sangre , ADP-Ribosil Ciclasa 1/genética , Subfamilia B de Transportador de Casetes de Unión a ATP/sangre , Subfamilia B de Transportador de Casetes de Unión a ATP/genética , Adiposidad , Adulto , Biomarcadores/sangre , Glucemia/metabolismo , Citocinas/sangre , Citocinas/genética , Femenino , Regulación de la Expresión Génica , Humanos , Insulina/sangre , Análisis de los Mínimos Cuadrados , Lípidos/sangre , Masculino , Glicoproteínas de Membrana/sangre , Glicoproteínas de Membrana/genética , NAD/sangre , NADP/sangre , Nicotinamida Fosforribosiltransferasa/sangre , Nicotinamida Fosforribosiltransferasa/genética , Nicotinamida-Nucleótido Adenililtransferasa/sangre , Nicotinamida-Nucleótido Adenililtransferasa/genética , Proyectos Piloto , Análisis de Componente Principal , Factores Sexuales , Sirtuinas/genética , Factores de Tiempo , Adulto JovenRESUMEN
Nicotinamide mononucleotide adenylyltransferase (NMN-AT) activity has not hitherto been demonstrated in human red blood cells, owing to its low activity. Since it is usually located in the nucleus, the possibility of finding it in human erythrocytes was excluded. Here we report the first demonstration and characterization of NMN-AT in human red blood cells, by an HPLC method. The enzyme is Mg2+ dependent, with a Km of 0.303 mM for nicotinamide mononucleotide and 0.103 mM for ATP, and a Vmax of 346 nmol g Hb-1 h-1. The crude preparation is also active on nicotinic acid mononucleotide, producing nicotinic acid adenine dinucleotide. NMN-AT activity is inhibited by nicotinic acid mononucleotide, and nicotinic acid adenylyltransferase is inhibited by nicotinamide mononucleotide. Fiftyfold purification of NMN-AT was achieved by DEAE-Toyopearl chromatography, and the kinetic characteristics were determined. The partially purified preparation maintained its nicotinic acid adenylyltransferase activity. These findings are discussed in light of the regulation of NAD metabolism in human red blood cells.