RESUMEN
In recent years, the potential of stem cell research for tissue engineering-based therapies and regenerative medicine clinical applications has become well established. In 2006, Chung pioneered the first entire organ transplant using adult stem cells and a scaffold for clinical evaluation. With this a new milestone was achieved, with seven patients with myelomeningocele receiving stem cell-derived bladder transplants resulting in substantial improvements in their quality of life. While a bladder is a relatively simple organ, the breakthrough highlights the incredible benefits that can be gained from the cross-disciplinary nature of tissue engineering and regenerative medicine (TERM) that encompasses stem cell research and stem cell bioprocessing. Unquestionably, the development of bioprocess technologies for the transfer of the current laboratory-based practice of stem cell tissue culture to the clinic as therapeutics necessitates the application of engineering principles and practices to achieve control, reproducibility, automation, validation and safety of the process and the product. The successful translation will require contributions from fundamental research (from developmental biology to the 'omics' technologies and advances in immunology) and from existing industrial practice (biologics), especially on automation, quality assurance and regulation. The timely development, integration and execution of various components will be critical-failures of the past (such as in the commercialization of skin equivalents) on marketing, pricing, production and advertising should not be repeated. This review aims to address the principles required for successful stem cell bioprocessing so that they can be applied deftly to clinical applications.
Asunto(s)
Medicina Regenerativa/métodos , Células Madre/citología , Ingeniería de Tejidos/métodos , Animales , Humanos , Andamios del TejidoAsunto(s)
Trastornos Distónicos/genética , Glicoproteínas de Membrana , Proteínas del Tejido Nervioso , Polimorfismo Genético , Receptores de Dopamina D1/genética , Adulto , Edad de Inicio , Anciano , Alelos , Blefaroespasmo/genética , Estudios de Casos y Controles , Mapeo Cromosómico , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática , Femenino , Predisposición Genética a la Enfermedad , Humanos , Masculino , Proteínas de Transporte de Membrana/genética , Repeticiones de Microsatélite/genética , Persona de Mediana Edad , Receptores de Dopamina D5 , Tortícolis/genéticaRESUMEN
We describe two affected individuals in a family with myoclonus-dystonia syndrome complicated with severe depression. One individual committed suicide. Molecular genetic analysis revealed a heterozygous point mutation in the epsilon-sarcoglycan gene, which we show leads to skipping of exon 5. This report suggests that the psychiatric spectrum of MDS includes more severe depression.
Asunto(s)
Antiparkinsonianos/uso terapéutico , Trastorno Depresivo Mayor/epidemiología , Trastorno Depresivo Mayor/psicología , Distonía , Exones/genética , Levodopa/uso terapéutico , Mioclonía , Mutación Puntual/genética , Sarcoglicanos/genética , Adulto , Anciano , Trastorno Depresivo Mayor/diagnóstico , Esquema de Medicación , Distonía/tratamiento farmacológico , Distonía/epidemiología , Distonía/genética , Expresión Génica/genética , Humanos , Incidencia , Masculino , Mioclonía/tratamiento farmacológico , Mioclonía/epidemiología , Mioclonía/genéticaRESUMEN
A single GAG deletion in the DYT1 gene causes primary early-onset, generalized torsion dystonia. The DYT1 protein product, torsinA, belongs to the AAA+ family of proteins. When overexpressed, wild-type torsinA localizes mainly to the endoplasmic reticulum, whereas the mutant forms inclusions of unclear biogenetic origin. In this study, overexpressed wild-type torsinA in human neuroblastoma (SH-SY5Y) cell lines was distributed throughout the cell body and colocalized with a marker for the endoplasmic reticulum, confirming it is an endoplasmic reticulum protein. However, mutant torsinA showed perinuclear staining and formed distinct globular inclusions, which did not colocalize with endoplasmic reticulum markers. Immunoelectron microscopy of the mutant torsinA inclusions revealed membrane whorls staining for torsinA, as well as labeling of lamellae, isolated bilayers, and perinuclear membranes. This finding shows that mutant torsinA redistributes to specific membranous structures, which may represent different stages of maturation of the intracellular inclusions. The mutant torsinA-containing bodies were immunoreactive for vesicular monoamine transporter 2 (VMAT2). VMAT2 expression is important for the exocytosis of bioactive monoamines in neurons. Abnormal processing, transport, or entrapment of VMAT2 within the mutant torsinA membranous inclusions, therefore, may affect cellular dopamine release, providing a potential pathogenic mechanism for the DYT1-dependent dystonia.
Asunto(s)
Distonía Muscular Deformante/genética , Distonía Muscular Deformante/metabolismo , Retículo Endoplásmico/metabolismo , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/metabolismo , Chaperonas Moleculares/genética , Chaperonas Moleculares/metabolismo , Neuroblastoma/genética , Neuroblastoma/metabolismo , Mutación Puntual/genética , Factores de Edad , Western Blotting , Línea Celular Tumoral , ADN Complementario/genética , Técnica del Anticuerpo Fluorescente , Eliminación de Gen , Humanos , Microscopía Electrónica , Neuroblastoma/patología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transfección , Repeticiones de Trinucleótidos/genética , Proteínas de Transporte Vesicular de Aminas Biógenas , Proteínas de Transporte Vesicular de MonoaminasRESUMEN
The epsilon-sarcoglycan gene (SGCE) on human chromosome 7q21 has been reported to be a major locus for inherited myoclonus-dystonia. Linkage to the SGCE locus has been detected in the majority of families tested, and mutations in the coding region have been found recently in families with autosomal dominant myoclonus-dystonia. To evaluate the relevance of SGCE in myoclonus-dystonia, we sequenced the entire coding region of the epsilon-sarcoglycan gene in 16 patients with either sporadic or familial myoclonus-dystonia. No mutations were found. This study suggests that epsilon-sarcoglycan does not play an important role in sporadic myoclonus-dystonia and supports genetic heterogeneity in familial cases.