RESUMEN
McArdle disease is a rare autosomal recessive disorder caused by mutations in the PYGM gene. This gene encodes for the skeletal muscle isoform of glycogen phosphorylase (myophosphorylase), the first enzyme in glycogenolysis. Patients with this disorder are unable to obtain energy from their glycogen stored in skeletal muscle, prompting an exercise intolerance. Currently, there is no treatment for this disease, and the lack of suitable in vitro human models has prevented the search for therapies against it. In this article, we have established the first human iPSC-based model for McArdle disease. For the generation of this model, induced pluripotent stem cells (iPSCs) from a patient with McArdle disease (harbouring the homozygous mutation c.148C>T; p.R50* in the PYGM gene) were differentiated into myogenic cells able to contract spontaneously in the presence of motor neurons and generate calcium transients, a proof of their maturity and functionality. Additionally, an isogenic skeletal muscle model of McArdle disease was created. As a proof-of-concept, we have tested in this model the rescue of PYGM expression by two different read-through compounds (PTC124 and RTC13). The developed model will be very useful as a platform for testing drugs or compounds with potential pharmacological activity.
Asunto(s)
Glucógeno Fosforilasa de Forma Muscular , Enfermedad del Almacenamiento de Glucógeno Tipo V , Células Madre Pluripotentes Inducidas , Humanos , Enfermedad del Almacenamiento de Glucógeno Tipo V/genética , Células Madre Pluripotentes Inducidas/metabolismo , Glucógeno/metabolismo , TecnologíaRESUMEN
The implementation of induced pluripotent stem cells (iPSCs) in biomedical research more than a decade ago, resulted in a huge leap forward in the highly promising area of personalized medicine. Nowadays, we are even closer to the patient than ever. To date, there are multiple examples of iPSCs applications in clinical trials and drug screening. However, there are still many obstacles to overcome. In this review, we will focus our attention on the advantages of implementing induced pluripotent stem cells technology into the clinics but also commenting on all the current drawbacks that could hinder this promising path towards the patient.
Asunto(s)
Diferenciación Celular , Células Madre Pluripotentes Inducidas/trasplante , Medicina de Precisión/tendencias , Humanos , Medicina de Precisión/métodosRESUMEN
A mouse iPSC line, IISHDOi005-A, generated from fibroblasts obtained from a mouse C57BL/6J with an age of 1â¯year and a half, has been obtained. For this purpose, reprogramming factors Oct3/4, Sox2, Klf4, and c-Myc were delivered using Sendai virus.
Asunto(s)
Línea Celular , Células Madre Pluripotentes Inducidas , Envejecimiento/patología , Animales , Diferenciación Celular , Técnicas de Reprogramación Celular , Fibroblastos , Cariotipo , Factor 4 Similar a Kruppel , Ratones Endogámicos C57BL , Virus SendaiRESUMEN
A human iPSC line, IISHDOi004-A, from fibroblasts obtained from a patient with Usher syndrome, harboring a homozygous mutation in the USH2A gene (c.2276G>T; p.Cys759Phe) has been generated. Reprogramming factors Oct3/4, Sox2, Klf4, and c-Myc were delivered using Sendai virus.
Asunto(s)
Proteínas de la Matriz Extracelular/genética , Células Madre Pluripotentes Inducidas/metabolismo , Síndromes de Usher/genética , Línea Celular , Humanos , Factor 4 Similar a Kruppel , MutaciónRESUMEN
We have generated a human iPSC line, IISHDOi002-A, from commercial primary normal human dermal fibroblasts belonging to an African mitochondrial haplogroup (L3), and with a 46, XY/47, XYY mosaicism. For this purpose, reprogramming factors Oct3/4, Sox2, Klf4 and cMyc were delivered using a non-integrative methodology that involves the use of Sendai virus.
Asunto(s)
Población Negra/genética , Técnicas de Cultivo de Célula/métodos , Cromosomas Humanos/genética , Haplotipos/genética , Mitocondrias/genética , Mosaicismo , Secuencia de Bases , Diferenciación Celular , Línea Celular , Humanos , Recién Nacido , Cariotipificación , Factor 4 Similar a Kruppel , Masculino , Mycoplasma/aislamiento & purificaciónRESUMEN
Human iPSC line IISHDOi001-A was generated from fibroblasts of a patient with McArdle disease harbouring the mutation, c.148C>T; p.Arg50Ter, in the PYGM gene. Reprogramming factors Oct3/4, Sox2, Klf4, and c-Myc were delivered using Sendai virus.
