RESUMEN
Crouzon syndrome is a debilitating congenital disorder involving abnormal craniofacial skeletal development caused by mutations in fibroblast growth factor receptor-2 (FGFR2). Phenotypic expression in humans exhibits an autosomal dominant pattern that commonly involves premature fusion of the coronal suture (craniosynostosis) and severe midface hypoplasia. To further investigate the biologic mechanisms by which the Crouzon syndrome-associated FGFR2(C342Y) mutation leads to abnormal craniofacial skeletal development, we created congenic BALB/c FGFR2(C342Y/+) mice. Here, we show that BALB/c FGFR2(C342Y/+) mice have a consistent craniofacial phenotype including partial fusion of the coronal and lambdoid sutures, intersphenoidal synchondrosis, and multiple facial bones, with minimal fusion of other craniofacial sutures. This phenotype is similar to the classic and less severe form of Crouzon syndrome that involves significant midface hypoplasia with limited craniosynostosis. Linear and morphometric analyses demonstrate that FGFR2(C342Y/+) mice on the BALB/c genetic background differ significantly in form and shape from their wild-type littermates and that in this genetic background the FGFR2(C342Y) mutation preferentially affects some craniofacial bones and sutures over others. Analysis of cranial bone cells indicates that the FGFR2(C342Y) mutation promotes aberrant osteoblast differentiation and increased apoptosis that is more severe in frontal than parietal bone cells. Additionally, FGFR2(C342Y/+) frontal, but not parietal, bones exhibit significantly diminished bone volume and density compared to wild-type mice. These results confirm that FGFR2-associated craniosynostosis occurs in association with diminished cranial bone tissue and may provide a potential biologic explanation for the clinical finding of phenotype consistency that exists between many Crouzon syndrome patients.
Asunto(s)
Huesos/fisiología , Mutación , Receptor Tipo 2 de Factor de Crecimiento de Fibroblastos/genética , Cráneo/patología , Animales , Apoptosis , Huesos/patología , Diferenciación Celular , Disostosis Craneofacial/genética , Craneosinostosis/genética , Craneosinostosis/fisiopatología , Modelos Animales de Enfermedad , Femenino , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Osteoblastos/patología , Fenotipo , Factores de TiempoRESUMEN
Microcephaly is a neurodevelopmental disorder causing significantly reduced cerebral cortex size. Many known microcephaly gene products localize to centrosomes, regulating cell fate and proliferation. Here, we identify and characterize a nuclear zinc finger protein, ZNF335/NIF-1, as a causative gene for severe microcephaly, small somatic size, and neonatal death. Znf335 null mice are embryonically lethal, and conditional knockout leads to severely reduced cortical size. RNA-interference and postmortem human studies show that ZNF335 is essential for neural progenitor self-renewal, neurogenesis, and neuronal differentiation. ZNF335 is a component of a vertebrate-specific, trithorax H3K4-methylation complex, directly regulating REST/NRSF, a master regulator of neural gene expression and cell fate, as well as other essential neural-specific genes. Our results reveal ZNF335 as an essential link between H3K4 complexes and REST/NRSF and provide the first direct genetic evidence that this pathway regulates human neurogenesis and neuronal differentiation.
Asunto(s)
Proteínas Portadoras/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Células-Madre Neurales/metabolismo , Neurogénesis , Proteínas Nucleares/metabolismo , Animales , Diferenciación Celular , Proliferación Celular , Proteínas de Unión al ADN , Femenino , Técnicas de Silenciamiento del Gen , Genes Letales , N-Metiltransferasa de Histona-Lisina , Humanos , Masculino , Ratones , Ratones Noqueados , Microcefalia/metabolismo , Complejos Multiproteicos/metabolismo , Proteína de la Leucemia Mieloide-Linfoide/metabolismo , Proteínas Represoras/metabolismo , Factores de TranscripciónRESUMEN
Pax3 and Pax7 paralogous genes have functionally diverged in vertebrate evolution, creating opportunity for a new distribution of roles between the two genes and the evolution of novel functions. Here we focus on the regulation and function of Pax7 in the brain and neural crest of amphibian embryos, which display a different pax7 expression pattern, compared to the other vertebrates already described. Pax7 expression is restricted to the midbrain, hindbrain and anterior spinal cord, and Pax7 activity is important for maintaining the fates of these regions, by restricting otx2 expression anteriorly. In contrast, pax3 displays broader expression along the entire neuraxis and Pax3 function is important for posterior brain patterning without acting on otx2 expression. Moreover, while both genes are essential for neural crest patterning, we show that they do so using two distinct mechanisms: Pax3 acts within the ectoderm which will be induced into neural crest, while Pax7 is essential for the inducing activity of the paraxial mesoderm towards the prospective neural crest.
Asunto(s)
Cresta Neural/metabolismo , Factor de Transcripción PAX7/metabolismo , Factores de Transcripción Paired Box/metabolismo , Proteínas de Xenopus/metabolismo , Xenopus laevis/embriología , Animales , Blastómeros/metabolismo , Tipificación del Cuerpo/genética , Ectodermo/metabolismo , Embrión no Mamífero/metabolismo , Embrión no Mamífero/fisiología , Inmunohistoquímica , Hibridación in Situ , Mesodermo/metabolismo , Microinyecciones , Modelos Biológicos , Sistema Nervioso/metabolismo , Oligonucleótidos Antisentido/farmacología , Técnicas de Cultivo de Órganos , Factor de Transcripción PAX3 , Factor de Transcripción PAX7/genética , Factores de Transcripción Paired Box/genética , ARN Mensajero/metabolismo , Proteínas de Xenopus/genéticaRESUMEN
PURPOSE: Osteonecrosis of the jaw (ONJ) has been observed recently in patients with cancer who are receiving intravenous bisphosphonate (BP) therapy. The incidence of BP-associated ONJ has not been well established. The purpose of this study was to determine the incidence of ONJ in a cohort of patients with multiple myeloma (MM), breast cancer (BC), or prostate cancer (PC) who were receiving BP therapy. PATIENTS AND METHODS: A retrospective chart review was performed. Medical record numbers were identified by ICD-9 codes: 203.0, 203.01, 174.9, and 185.0 for active MM, MM in remission, BC, and PC, respectively. Patients were included if they were evaluated and/or treated between January 1, 2000, and December 31, 2005, and had received zoledronic acid and/or pamidronate. Patients were excluded if they had a history of radiation therapy to the jaws or of tumors or cysts. ONJ was defined as clinical evidence of "exposed necrotic bone" in the mouth. RESULTS: Through evaluation of 1,086 patient medical records, it was determined that 447 subjects met the inclusion criteria: 11 of 292 patients with MM (3.8%; 95% confidence interval [CI], 1.6%, 6.0%) had ONJ, as did 2.5% of 81 patients with BC (0%, 6.9%) and 2.9% of 69 patients with PC (0%, 5.9%). CONCLUSION: The incidence of ONJ associated with intravenous BPs was at least 3.8 per 100 patients with MM, 2.5 per 100 patients with BC, and 2.9 per 100 patients with PC during the 5-year study period. The next phase of this study involves assessment of risk factors that differentiate these patients from those treated with BPs who do not develop ONJ.
Asunto(s)
Conservadores de la Densidad Ósea/efectos adversos , Difosfonatos/efectos adversos , Enfermedades Maxilomandibulares/inducido químicamente , Osteonecrosis/inducido químicamente , Conservadores de la Densidad Ósea/administración & dosificación , Neoplasias de la Mama/tratamiento farmacológico , Difosfonatos/administración & dosificación , Femenino , Humanos , Incidencia , Inyecciones Intravenosas , Enfermedades Maxilomandibulares/epidemiología , Masculino , Mieloma Múltiple/tratamiento farmacológico , Osteonecrosis/epidemiología , Neoplasias de la Próstata/tratamiento farmacológico , Estudios RetrospectivosRESUMEN
TERT, the protein component of telomerase, serves to maintain telomere function through the de novo addition of telomere repeats to chromosome ends, and is reactivated in 90% of human cancers. In normal tissues, TERT is expressed in stem cells and in progenitor cells, but its role in these compartments is not fully understood. Here we show that conditional transgenic induction of TERT in mouse skin epithelium causes a rapid transition from telogen (the resting phase of the hair follicle cycle) to anagen (the active phase), thereby facilitating robust hair growth. TERT overexpression promotes this developmental transition by causing proliferation of quiescent, multipotent stem cells in the hair follicle bulge region. This new function for TERT does not require the telomerase RNA component, which encodes the template for telomere addition, and therefore operates through a mechanism independent of its activity in synthesizing telomere repeats. These data indicate that, in addition to its established role in extending telomeres, TERT can promote proliferation of resting stem cells through a non-canonical pathway.
Asunto(s)
Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Folículo Piloso/citología , Células Madre/citología , Telomerasa/genética , Telomerasa/metabolismo , Animales , Bromodesoxiuridina/metabolismo , Proliferación Celular , Proteínas de Unión al ADN/deficiencia , Inducción Enzimática , Folículo Piloso/enzimología , Ratones , Ratones Transgénicos , ARN/genética , Células Madre/enzimología , Telomerasa/deficiencia , Transgenes/genéticaRESUMEN
FGF, WNT, and BMP signaling promote neural crest formation at the neural plate boundary in vertebrate embryos. To understand how these signals are integrated, we have analyzed the role of the transcription factors Msx1 and Pax3. Using a combination of overexpression and morpholino-mediated knockdown strategies in Xenopus, we show that Msx1 and Pax3 are both required for neural crest formation, display overlapping but nonidentical activities, and that Pax3 acts downstream of Msx1. In neuralized ectoderm, Msx1 is sufficient to induce multiple early neural crest genes. Msx1 induces Pax3 and ZicR1 cell autonomously, in turn, Pax3 combined with ZicR1 activates Slug in a WNT-dependent manner. Upstream of this, WNTs initiate Slug induction through Pax3 activity, whereas FGF8 induces neural crest through both Msx1 and Pax3 activities. Thus, WNT and FGF8 signals act in parallel at the neural border and converge on Pax3 activity during neural crest induction.