RESUMEN
Autoimmune and autoinflammatory diseases (AIDs) are a heterogeneous group of diseases. They can occur in childhood and account for significant morbidity and mortality. Transitioning from pediatric to adult healthcare can be difficult for patients and their families. It can interfere with patient follow-up and management, and eventually lead to complications. Although recommendations exist for the successful transition of patients with chronic diseases, few are specifically adapted to children and adults with AIDs (Suris et al., 2015-Solau-Gervais, 2012). The French working group on transition of the rare autoimmune and autoinflammatory diseases presents its reflections and recommendations for a successful transition. Preparation for transition should start early. Its goals are to empower adolescents by providing them with the knowledge to manage their own care, respond appropriately to changes in their condition, and evolve within the adult healthcare system. This requires the active participation of the patient, his or her family, as well as the pediatric and adult medical teams. The transition process involves multidisciplinary care and dedicated therapeutic education programs. Finally, the identification of medical specialists by region, trained in rare AIDs and accompanied by expert patients, may improve the management of patients with rare AIDs from adolescence to adulthood.
Asunto(s)
Enfermedades Autoinflamatorias Hereditarias , Transición a la Atención de Adultos , Adolescente , Adulto , Niño , Femenino , Enfermedades Autoinflamatorias Hereditarias/diagnóstico , Enfermedades Autoinflamatorias Hereditarias/epidemiología , Enfermedades Autoinflamatorias Hereditarias/terapia , Humanos , Masculino , Enfermedades RarasRESUMEN
Calpain 3 is known as the skeletal muscle-specific member of the calpains, a family of intracellular nonlysosomal cysteine proteases. It was previously shown that defects in the human calpain 3 gene are responsible for limb girdle muscular dystrophy type 2A (LGMD2A), an inherited disease affecting predominantly the proximal limb muscles. To better understand the function of calpain 3 and the pathophysiological mechanisms of LGMD2A and also to develop an adequate model for therapy research, we generated capn3-deficient mice by gene targeting. capn3-deficient mice are fully fertile and viable. Allele transmission in intercross progeny demonstrated a statistically significant departure from Mendel's law. capn3-deficient mice show a mild progressive muscular dystrophy that affects a specific group of muscles. The age of appearance of myopathic features varies with the genetic background, suggesting the involvement of modifier genes. Affected muscles manifest a similar apoptosis-associated perturbation of the IkappaBalpha/nuclear factor kappaB pathway as seen in LGMD2A patients. In addition, Evans blue staining of muscle fibers reveals that the pathological process due to calpain 3 deficiency is associated with membrane alterations.
Asunto(s)
Apoptosis , Calpaína/deficiencia , Proteínas de Unión al ADN/metabolismo , Proteínas I-kappa B , Distrofias Musculares/enzimología , Distrofias Musculares/metabolismo , FN-kappa B/metabolismo , Transducción de Señal , Animales , Calpaína/química , Calpaína/genética , Calpaína/metabolismo , Creatina Quinasa/metabolismo , Cruzamientos Genéticos , Azul de Evans , Femenino , Fertilidad , Eliminación de Gen , Marcación de Gen , Genotipo , Masculino , Ratones , Ratones Noqueados , Fibras Musculares Esqueléticas/enzimología , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/patología , Músculo Esquelético/enzimología , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Distrofias Musculares/genética , Distrofias Musculares/patología , Inhibidor NF-kappaB alfa , Fenotipo , ARN Mensajero/análisis , ARN Mensajero/genética , Sarcolema/patologíaRESUMEN
Calpain 3 is a nonlysosomal cysteine protease whose biological functions remain unknown. We previously demonstrated that this protease is altered in limb girdle muscular dystrophy type 2A patients. Preliminary observations suggested that its gene is subjected to alternative splicing. In this paper, we characterize transcriptional and posttranscriptional events leading to alterations involving the NS, IS1, and IS2 regions and/or the calcium binding domains of the mouse calpain 3 gene (capn3). These events can be divided into three groups: (i) splicing of exons that preserve the translation frame, (ii) inclusion of two distinct intronic sequences between exons 16 and 17 that disrupt the frame and would lead, if translated, to a truncated protein lacking domain IV, and (iii) use of an alternative first exon specific to lens tissue. In addition, expression of these isoforms seems to be regulated. Investigation of the proteolytic activities and titin binding abilities of the translation products of some of these isoforms clearly indicated that removal of these different protein segments affects differentially the biochemical properties examined. In particular, removal of exon 6 impaired the autolytic but not fodrinolytic activity and loss of exon 16 led to an increased titin binding and a loss of fodrinolytic activity. These results are likely to impact our understanding of the pathophysiology of calpainopathies and the development of therapeutic strategies.
Asunto(s)
Calpaína/genética , Calpaína/metabolismo , Isoenzimas , Procesamiento Postranscripcional del ARN , Transcripción Genética , Empalme Alternativo , Animales , Encéfalo/metabolismo , Proteínas Portadoras/metabolismo , Células Cultivadas , Clonación Molecular , Conectina , Cartilla de ADN , Embrión de Mamíferos/anatomía & histología , Embrión de Mamíferos/metabolismo , Humanos , Hibridación in Situ , Intrones , Cristalino/anatomía & histología , Cristalino/metabolismo , Ratones , Ratones Endogámicos BALB C , Proteínas de Microfilamentos/metabolismo , Modelos Genéticos , Proteínas Musculares/metabolismo , Músculo Esquelético/metabolismo , Músculo Liso/metabolismo , Miocardio/metabolismo , Fragmentos de Péptidos/metabolismo , Proteínas Quinasas/metabolismo , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Distribución TisularRESUMEN
We analysed the clinical, histochemical, ultrastructural and genetic data of patients affected by central core disease (CCD) studied during the last 20 years. From a total series of 86 CCD-families, we have identified 46 CCD families with RYR1 mutations (16 autosomal dominant, 8 autosomal recessive, 17 sporadic cases and 5 de novo mutations). Out of the other 40 CCD families, the RyR1 gene was entirely excluded in 7 families, by cDNA sequencing or linkage analysis, indicating a genetic heterogeneity of CCD.
Asunto(s)
Miopatía del Núcleo Central/diagnóstico , Miopatía del Núcleo Central/genética , Canal Liberador de Calcio Receptor de Rianodina/genética , Humanos , Inmunohistoquímica , Miopatía del Núcleo Central/patologíaRESUMEN
Hypophosphatasia is an inherited disorder characterized by defective bone mineralization and deficiency of serum and tissue liver/bone/kidney alkaline phosphatase (L/B/K ALP) activity. We report the characterization of ALPL gene mutations in a series of 11 families from various origins affected by perinatal and infantile hypophosphatasia. Sixteen distinct mutations were found, fifteen of them not previously reported: M45V, G46R, 388-391delGTAA, 389delT, T131I, G145S, D172E, 662delG, G203A, R255L, 876-881delAGGGGA, 962delG, E294K, E435K, and A451T. This confirms that severe hypophosphatasia is due to a large spectrum of mutations in Caucasian populations.
Asunto(s)
Fosfatasa Alcalina/genética , Hipofosfatasia/enzimología , Hipofosfatasia/genética , Mutación , Femenino , Humanos , Hipofosfatasia/diagnóstico , Recién Nacido , Masculino , Tamizaje Neonatal , Embarazo , Diagnóstico PrenatalAsunto(s)
Fosfatasa Alcalina/genética , Caries Dental/genética , Tamización de Portadores Genéticos , Hipofosfatasia/genética , Mutación , Enfermedades Dentales/genética , Adulto , Fosfatasa Alcalina/sangre , Fosfatasa Alcalina/deficiencia , Animales , Células COS , Línea Celular , Niño , Preescolar , Chlorocebus aethiops , Caries Dental/sangre , Caries Dental/enzimología , Femenino , Humanos , Hipofosfatasia/sangre , Hipofosfatasia/enzimología , Masculino , Modelos Moleculares , Mutagénesis Sitio-Dirigida/genética , Mutación Missense/genética , Especificidad de Órganos/genética , Linaje , Estructura Cuaternaria de Proteína/genética , Enfermedades Dentales/sangre , Enfermedades Dentales/enzimologíaRESUMEN
OBJECTIVE: To characterize the muscle involvement of patients with central core disease (CCD) caused by mutations in the ryanodine receptor 1 gene (RYR1) and to compare these findings with those from patients with core myopathies unlinked to the RYR1 gene. METHODS: We performed a systematic muscular imaging assessment in 11 patients with an RYR1 gene mutation and compared these findings with those of 5 patients from two unrelated families with autosomal dominant core myopathies not linked to RYR1, ACTA1, or MYH7 gene loci. RESULTS: All patients with RYR1 CCD had a characteristic pattern with predominant involvement of the gluteus maximus, adductor magnus, sartorius, vastus intermediolateralis, soleus, and lateral gastrocnemius muscles. In contrast, muscle CT in the first family not linked to RYR1 showed predominant affection of the gluteus minimus and hamstring muscles, whereas the second family presented with predominant involvement of the gluteus minimus, vastus intermediolateralis, tibialis anterior, and medial gastrocnemius muscles. In addition to muscle imaging data, we present detailed information on the clinical and pathologic findings of these novel phenotypes of core myopathies not linked to RYR1. CONCLUSIONS: Our data suggest genetic heterogeneity in autosomal dominant core myopathies and the existence of additional unidentified genes.
Asunto(s)
Trastornos de los Cromosomas/genética , Trastornos de los Cromosomas/patología , Músculo Esquelético/patología , Miopatía del Núcleo Central/genética , Miopatía del Núcleo Central/patología , Canal Liberador de Calcio Receptor de Rianodina/genética , Predisposición Genética a la Enfermedad/genética , Humanos , Estadística como AsuntoRESUMEN
Lack of functional calpain 3 in humans is a cause of limb girdle muscular dystrophy, but the function(s) of calpain 3 remain(s) unknown. Special muscle conditions in which calpain 3 is downregulated could yield valuable clues to the understanding of its function(s). We monitored calpain 3 mRNA amounts by quantitative RT-PCR and compared them with those of alpha-skeletal actin mRNA in mouse leg muscles for different types of denervation and muscle injury. Intact muscle denervation reduced calpain 3 mRNA expression by a factor of 5 to 10, while alpha-skeletal actin mRNA was reduced in a slower and less extensive manner. Muscle injury (denervation-devascularization), which leads to muscle degeneration and regeneration, induced a 20-fold decrease in the mRNA level of both calpain 3 and alpha-skeletal actin. Furthermore, whereas in normal muscle and intact denervated muscle, the full-length transcript is the major calpain 3 mRNA, in injured muscle, isoforms lacking exon 6 are predominant during the early regeneration process. These data suggest that muscle condition determines the specific calpain 3 isoform pattern of expression and that calpain 3 expression is downregulated by denervation.
Asunto(s)
Calpaína/genética , Regulación Enzimológica de la Expresión Génica/fisiología , Proteínas Musculares , Músculo Esquelético/fisiología , Regeneración/fisiología , Actinas/genética , Empalme Alternativo/fisiología , Animales , Apoptosis/fisiología , Cartilla de ADN , Masculino , Ratones , Desnervación Muscular , Fibras Musculares Esqueléticas/enzimología , Fibras Musculares Esqueléticas/patología , Músculo Esquelético/patología , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Nervio Ciático/fisiología , Nervio Ciático/cirugíaRESUMEN
Limb-girdle muscular dystrophy, type 2A (LGMD 2A), is an autosomal recessive disorder that causes late-onset muscle-wasting, and is due to mutations in the muscle-specific protease calpain 3 (C3). Although LGMD 2A would be a feasible candidate for gene therapy, the reported instability of C3 in vitro raised questions about the potential of obtaining a stable, high-level expression of C3 from a transgene in vivo. We have generated transgenic (Tg) mice with muscle-specific overexpression of full-length C3 or C3 isoforms, which arise from alternative splicing, to test whether stable expression of C3 transgenes could occur in vivo. Unexpectedly, we found that full-length C3 can be overexpressed at high levels in vivo, without toxicity. In addition, we found that Tg expressing C3 lacking exon 6, an isoform expressed embryonically, have muscles that resemble regenerating or developing muscle. Tg expressing C3 lacking exon 15 shared this morphology in the soleus, but not other muscles. Assays of inflammation or muscle membrane damage indicated that the Tg muscles were not degenerative, suggesting that the immature muscle resulted from a developmental block rather than degeneration and regeneration. These studies show that C3 can be expressed stably in vivo from a transgene, and indicate that alternatively spliced C3 isoforms should not be used in gene-therapy applications because they impair proper muscle development.
Asunto(s)
Calpaína/genética , Músculo Esquelético/crecimiento & desarrollo , Transgenes , Animales , Apoptosis , Secuencia de Bases , Cartilla de ADN , Humanos , Inmunohistoquímica , Ratones , Ratones Transgénicos , Músculo Esquelético/metabolismoRESUMEN
Our understanding of early human development has been impeded by the general difficulty in obtaining suitable samples for study. As a result, and because of the extraordinarily high degree of evolutionary conservation of many developmentally important genes and developmental pathways, great reliance has been placed on extrapolation from animal models of development, principally the mouse. However, the strong evolutionary conservation of coding sequence for developmentally important genes does not necessarily mean that their expression patterns are as highly conserved. The very recent availability of human embryonic samples for gene expression studies has now permitted for the first time an assessment of the degree to which we can confidently extrapolate from studies of rodent gene expression patterns. We have found significant human-mouse differences in embryonic expression patterns for a variety of genes. We present detailed data for two illustrative examples. Wnt7a, a very highly conserved gene known to be important in early development, shows significant differences in spatial and temporal expression patterns in the developing brain (midbrain, telencephalon) of man and mice. CAPN3, the locus for LGMD2A limb girdle muscular dystrophy, and its mouse orthologue differ extensively in expression in embryonic heart, lens and smooth muscle. Our study also shows how molecular analyses, while providing explanations for the observed differences, can be important in providing insights into mammalian evolution.