RESUMEN
Germinal centers (GCs) are sites of intense B cell proliferation and are central for T cell-dependent antibody responses. However, the role of c-Myc, a key cell-cycle regulator, in this process has been questioned. Here we identified c-Myc(+) B cell subpopulations in immature and mature GCs and found, by genetic ablation of Myc, that they had indispensable roles in the formation and maintenance of GCs. The identification of these functionally critical cellular subsets has implications for human B cell lymphomagenesis, which originates mostly from GC B cells and frequently involves MYC chromosomal translocations. As these translocations are generally dependent on transcription of the recombining partner loci, the c-Myc(+) GC subpopulations may be at a particularly high risk for malignant transformation.
Asunto(s)
Subgrupos de Linfocitos B/inmunología , Linfocitos B/metabolismo , Ciclo Celular/genética , Centro Germinal/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , Animales , Linfocitos B/inmunología , Linfocitos B/patología , Ciclo Celular/inmunología , Proliferación Celular , Transformación Celular Neoplásica/genética , Transformación Celular Neoplásica/inmunología , Eliminación de Gen , Regulación de la Expresión Génica/inmunología , Genes Reporteros , Sitios Genéticos , Centro Germinal/inmunología , Centro Germinal/patología , Proteínas Fluorescentes Verdes , Linfoma/genética , Linfoma/metabolismo , Linfoma/patología , Ratones , Ratones Transgénicos , Proteínas Proto-Oncogénicas c-myc/deficiencia , Proteínas Proto-Oncogénicas c-myc/inmunología , Transducción de Señal/genética , Transducción de Señal/inmunología , Linfocitos T/inmunología , Linfocitos T/metabolismo , Linfocitos T/patología , Translocación GenéticaRESUMEN
Since the discovery of the myc gene, few genes are likely to have such influence on biomedical research. The diversity of the biological functions regulated by this transcription factor and its impact in human health have attracted investigators from many different fields. The development of conditional knockout mouse models has allowed for the characterization of Myc-driven molecular mechanisms in primary cells in physiological and pathological conditions. In this review, we discuss some of the main functions and recent findings regarding c-Myc in in vivo B lymphocyte differentiation from early progenitors to terminally differentiated cells.
Asunto(s)
Linfocitos B/citología , Linfocitos B/metabolismo , Diferenciación Celular , Proteínas Proto-Oncogénicas c-myc/metabolismo , Animales , Humanos , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas c-myc/deficiencia , Proteínas Proto-Oncogénicas c-myc/genéticaRESUMEN
Inhibition of the mutationally activated Wnt cascade in colorectal cancer cell lines induces a rapid G1 arrest and subsequent differentiation. This arrest can be overcome by maintaining expression of a single Tcf4 target gene, the proto-oncogene c-Myc. Since colorectal cancer cells share many molecular characteristics with proliferative crypt progenitors, we have assessed the physiological role of c-Myc in adult crypts by conditional gene deletion. c-Myc-deficient crypts are lost within weeks and replaced by c-Myc-proficient crypts through a fission process of crypts that have escaped gene deletion. Although c-Myc(-/-) crypt cells remain in the cell cycle, they are on average much smaller than wild-type cells, cycle slower, and divide at a smaller cell size. c-Myc appears essential for crypt progenitor cells to provide the necessary biosynthetic capacity to successfully progress through the cell cycle.
Asunto(s)
Neoplasias Colorrectales/metabolismo , Genes myc , Intestinos/patología , Proteínas Proto-Oncogénicas c-myc/genética , Factores de Transcripción TCF/genética , Proteínas Wnt/genética , Animales , Apoptosis , Recuento de Células , Neoplasias Colorrectales/patología , Epitelio/fisiología , Marcación de Gen , Integrasas/genética , Mucosa Intestinal/metabolismo , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Mitosis , Proteína 2 Similar al Factor de Transcripción 7RESUMEN
OBJECTIVE: The c-Myc protein is a member of the basic region/helix-loop-helix/leucine zipper (bHLHZip) transcription factor family, which is implicated in regulation of proliferation, differentiation, and apoptosis in multiple cell types. The aim of this study was to characterize the role of the proto-oncogene c-myc in hematopoietic stem cells (HSC) during postnatal development. MATERIAL AND METHODS: We have generated a conditional mouse model that allows us to inactivate c-myc in bone marrow (BM) in an inducible fashion. RESULTS: We show that conditional inactivation of c-Myc in BM severely impairs HSC differentiation, leading to a striking decrease in the number of lymphoid and myeloid cells. c-Myc deletion in BM causes substantial accumulation of a Lin(-)Sca-1(+)c-Kit(-) cell population expressing high levels of the cell-cycle inhibitor p21, whose origin and function are otherwise poorly characterized. In vivo inactivation of p21 and c-Myc normalizes Lin(-)Sca-1(+)c-Kit(-) cell numbers and restores normal proliferation. The potential origin and function of these cells are discussed. CONCLUSIONS: c-Myc plays a role in HSC maintenance and differentiation and might be regulating generation of Lin(-)Sca-1(+)c-Kit(-) through the cell-cycle regulator p21.
Asunto(s)
Antígenos de Diferenciación/biosíntesis , Diferenciación Celular/fisiología , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Células Madre Hematopoyéticas/citología , Modelos Animales , Proteínas Proto-Oncogénicas c-myc/fisiología , Animales , Apoptosis , Proliferación Celular , Eliminación de Gen , Células Madre Hematopoyéticas/efectos de los fármacos , Humanos , Linfocitos/citología , Ratones , Células Mieloides/citología , Proto-Oncogenes Mas , Proteínas Proto-Oncogénicas c-kit/análisis , Proteínas Proto-Oncogénicas c-myc/genéticaRESUMEN
BCL6 protects germinal center (GC) B cells against DNA damage-induced apoptosis during somatic hypermutation and class-switch recombination. Although expression of BCL6 was not found in early IL-7-dependent B cell precursors, we report that IL-7Ralpha-Stat5 signaling negatively regulates BCL6. Upon productive VH-DJH gene rearrangement and expression of a mu heavy chain, however, activation of pre-B cell receptor signaling strongly induces BCL6 expression, whereas IL-7Ralpha-Stat5 signaling is attenuated. At the transition from IL-7-dependent to -independent stages of B cell development, BCL6 is activated, reaches expression levels resembling those in GC B cells, and protects pre-B cells from DNA damage-induced apoptosis during immunoglobulin (Ig) light chain gene recombination. In the absence of BCL6, DNA breaks during Ig light chain gene rearrangement lead to excessive up-regulation of Arf and p53. As a consequence, the pool of new bone marrow immature B cells is markedly reduced in size and clonal diversity. We conclude that negative regulation of Arf by BCL6 is required for pre-B cell self-renewal and the formation of a diverse polyclonal B cell repertoire.
Asunto(s)
Linfocitos B/citología , Linfocitos B/inmunología , Proteínas de Unión al ADN/inmunología , Factores de Ribosilacion-ADP/metabolismo , Animales , Apoptosis , Secuencia de Bases , Proliferación Celular , Supervivencia Celular , Células Cultivadas , Citoprotección , Daño del ADN/genética , Regulación hacia Abajo/genética , Reordenamiento Génico de Cadena Ligera de Linfocito B/genética , Humanos , Interleucina-7/metabolismo , Linfopoyesis , Ratones , Datos de Secuencia Molecular , Receptores de Células Precursoras de Linfocitos B/metabolismo , Células Precursoras de Linfocitos B/citología , Células Precursoras de Linfocitos B/metabolismo , Proteínas Proto-Oncogénicas c-bcl-6 , Proteínas Proto-Oncogénicas c-myc/metabolismo , Recombinación Genética/genética , Transducción de Señal , Transcripción Genética , Regulación hacia Arriba/genéticaRESUMEN
Cell proliferation is strictly controlled during differentiation. In T cell development, the cell cycle is normally arrested at the CD4(+)CD8(+) stage, but the mechanism underlying such differentiation-specific exit from the cell cycle has been unclear. Fbxw7 (also known as Fbw7, Sel-10, hCdc4, or hAgo), an F-box protein subunit of an SCF-type ubiquitin ligase complex, induces the degradation of positive regulators of the cell cycle, such as c-Myc, c-Jun, cyclin E, and Notch. FBXW7 is often mutated in a subset of human cancers. We have now achieved conditional inactivation of Fbxw7 in the T cell lineage of mice and found that the cell cycle is not arrested at the CD4(+)CD8(+) stage in the homozygous mutant animals. The mutant mice manifested thymic hyperplasia as a result of c-Myc accumulation and eventually developed thymic lymphoma. In contrast, mature T cells of the mutant mice failed to proliferate in response to mitogenic stimulation and underwent apoptosis in association with accumulation of c-Myc and p53. These latter abnormalities were corrected by deletion of p53. Our results suggest that Fbxw7 regulates the cell cycle in a differentiation-dependent manner, with its loss resulting in c-Myc accumulation that leads to hyperproliferation in immature T cells but to p53-dependent cell-cycle arrest and apoptosis in mature T cells.
Asunto(s)
Ciclo Celular , Diferenciación Celular , Proteínas F-Box/genética , Proteínas F-Box/metabolismo , Linfoma/patología , Mutación/genética , Linfocitos T/citología , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo , Animales , Antígenos/farmacología , Ciclo Celular/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Linaje de la Célula/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Susceptibilidad a Enfermedades , Proteína 7 que Contiene Repeticiones F-Box-WD , Regulación de la Expresión Génica/efectos de los fármacos , Integrasas/metabolismo , Ratones , Ratones Desnudos , Fenotipo , Proteínas Proto-Oncogénicas c-myc/metabolismo , Receptores Notch/metabolismo , Bazo/citología , Bazo/efectos de los fármacos , Linfocitos T/efectos de los fármacos , Linfocitos T/metabolismo , Timo/efectos de los fármacos , Timo/patología , Proteína p53 Supresora de Tumor/metabolismo , Ubiquitina-Proteína Ligasas/deficienciaRESUMEN
Deregulated expression of the Myc family of transcription factors (c-, N-, and L-myc) contributes to the development of many cancers by a mechanism believed to involve the stimulation of cell proliferation and inhibition of differentiation. However, using B cell-specific c-/N-myc double-knockout mice and E(mu)-myc transgenic mice bred onto genetic backgrounds (recombinase-activating gene 2-/- and Btk-/- Tec-/-) whereby B cell development is arrested, we show that Myc is necessary to stimulate both proliferation and differentiation in primary B cells. Moreover, Myc expression results in sustained increases in intracellular Ca2+ ([Ca2+]i), which is required for Myc to stimulate B cell proliferation and differentiation. The increase in [Ca2+]i correlates with constitutive nuclear factor of activated T cells (NFAT) nuclear translocation, reduced Ca2+ efflux, and decreased expression of the plasma membrane Ca2+-adenosine triphosphatase (PMCA) efflux pump. Our findings demonstrate a revised model whereby Myc promotes both proliferation and differentiation, in part by a remarkable mechanism whereby Myc amplifies Ca2+ signals, thereby enabling the concurrent expression of Myc- and Ca2+-regulated target genes.
Asunto(s)
Linfocitos B/metabolismo , Señalización del Calcio/fisiología , Genes myc , Proteínas Proto-Oncogénicas c-myc/fisiología , Animales , Linfocitos B/citología , Calcio/análisis , Calcio/metabolismo , Diferenciación Celular , Núcleo Celular/metabolismo , Células Cultivadas , Inmunoprecipitación de Cromatina , Fibroblastos/metabolismo , Genes Reporteros , Luciferasas/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Modelos Biológicos , Factores de Transcripción NFATC/metabolismo , ATPasas Transportadoras de Calcio de la Membrana Plasmática/metabolismoRESUMEN
c-Myc (Myc) is highly expressed in developing embryos where it regulates body size by controlling proliferation but not cell size. However, Myc is also induced in many postmitotic tissues, including adult myocardium, in response to stress where the predominant form of growth is an increase in cell size (hypertrophy) and not number. The function of Myc induction in this setting is unproven. Therefore, to explore Myc's role in hypertrophic growth, we created mice where Myc can be inducibly inactivated, specifically in adult myocardium. Myc-deficient hearts demonstrated attenuated stress-induced hypertrophic growth, secondary to a reduction in cell growth of individual myocytes. To explore the dependence of Myc-induced cell growth on CycD2, we created bigenic mice where Myc can be selectively activated in CycD2-null adult myocardium. Myc-dependent hypertrophic growth and cell cycle reentry is blocked in CycD2-deficient hearts. However, in contrast to Myc-induced DNA synthesis, hypertrophic growth is independent of CycD2-induced Cdk2 activity. These data suggest that Myc is required for a normal hypertrophic response and that its growth-promoting effects are also mediated through a CycD2-dependent pathway.
Asunto(s)
Cardiomegalia/patología , Ciclinas/fisiología , Miocitos Cardíacos/patología , Miocitos Cardíacos/fisiología , Proteínas Proto-Oncogénicas c-myc/fisiología , Animales , Apoptosis , Cardiomegalia/metabolismo , Ciclo Celular , Aumento de la Célula , Proliferación Celular , Células Cultivadas , Ciclina D2 , Ciclinas/genética , Isoproterenol/farmacología , Ratones , Ratones Transgénicos , Miocardio/metabolismo , Miocardio/patología , Miocitos Cardíacos/efectos de los fármacos , Proteínas Proto-Oncogénicas c-myc/genéticaRESUMEN
We examined the genetic requirements for the Myc family of oncogenes in normal Sonic hedgehog (Shh)-mediated cerebellar granule neuronal precursor (GNP) expansion and in Shh pathway-induced medulloblastoma formation. In GNP-enriched cultures derived from N-myc(Fl/Fl) and c-myc(Fl/Fl) mice, disruption of N-myc, but not c-myc, inhibited the proliferative response to Shh. Conditional deletion of c-myc revealed that, although it is necessary for the general regulation of brain growth, it is less important for cerebellar development and GNP expansion than N-myc. In vivo analysis of compound mutants carrying the conditional N-myc null and the activated Smoothened (ND2:SmoA1) alleles showed, that although granule cells expressing the ND2:SmoA1 transgene are present in the N-myc null cerebellum, no hyperproliferation or tumor formation was detected. Taken together, these findings provide in vivo evidence that N-myc acts downstream of Shh/Smo signaling during GNP proliferation and that N-myc is required for medulloblastoma genesis even in the presence of constitutively active signaling from the Shh pathway.
Asunto(s)
Neoplasias Cerebelosas/genética , Cerebelo/citología , Genes myc , Proteínas Hedgehog/fisiología , Meduloblastoma/genética , Animales , División Celular , Neoplasias Cerebelosas/patología , Cerebelo/patología , Niño , Humanos , Cinética , Meduloblastoma/patología , Ratones , Ratones Noqueados , Proteínas Proto-Oncogénicas c-myc/deficiencia , Proteínas Proto-Oncogénicas c-myc/genética , Transducción de SeñalRESUMEN
The transcription factor Myc (c-Myc) plays an important role in cell growth and cell death, yet its physiological function remains unclear. Ectopic activation of Myc has been recently suggested to regulate cell mass, and Drosophila dmyc controls cellular growth and size independently of cell division. By contrast, it has been proposed that in mammals Myc controls cell division and cell number. To gain insights into this debate we have specifically knocked out Myc in epidermis. Myc epidermal knockout mice are viable and their keratinocytes continue to cycle, but they display severe skin defects. The skin is tight and fragile, tears off in areas of mechanical friction and displays impaired wound healing. Steady-state epidermis is thinner, with loss of the proliferative compartment and premature differentiation. Remarkably, keratinocyte cell size, growth and endoreplication are reduced, and stem cell amplification is compromised. The results provide new and direct evidence for a role for endogenous Myc in cellular growth that is required for hyperproliferative cycles and tissue homeostasis.
Asunto(s)
Diferenciación Celular/fisiología , Epidermis/anomalías , Queratinocitos/metabolismo , Proteínas Proto-Oncogénicas c-myc/genética , Células Madre/metabolismo , Animales , Ciclo Celular/fisiología , División Celular/fisiología , Aumento de la Célula , Movimiento Celular/fisiología , Proliferación Celular , Tamaño de la Célula , Epidermis/patología , Epidermis/fisiopatología , Femenino , Folículo Piloso/anomalías , Folículo Piloso/patología , Folículo Piloso/fisiopatología , Homeostasis/fisiología , Queratinocitos/patología , Masculino , Ratones , Ratones Noqueados , Ratones Transgénicos , Ploidias , Células Madre/citología , Estrés Mecánico , Cicatrización de Heridas/fisiologíaRESUMEN
The c-Myc protein is a transcription factor implicated in the regulation of multiple biological processes, including cell proliferation, cell growth, and apoptosis. In vivo overexpression of c-myc is linked to tumor development in a number of mouse models. Here, we show that perinatal inactivation of c-Myc in liver causes disorganized organ architecture, decreased hepatocyte size, and cell ploidy. Furthermore, c-Myc appears to have distinct roles in proliferation in liver. Thus, postnatal hepatocyte proliferation does not require c-Myc, whereas it is necessary for liver regeneration in adult mice. These results show novel physiological functions of c-myc in liver development and hepatocyte proliferation and growth.
Asunto(s)
Proliferación Celular , Tamaño de la Célula , Hepatocitos/metabolismo , Ploidias , Proteínas Proto-Oncogénicas c-myc/deficiencia , Proteínas Proto-Oncogénicas c-myc/metabolismo , Animales , Bromodesoxiuridina , Cartilla de ADN , Fluoresceínas , Inmunohistoquímica , Hígado/crecimiento & desarrollo , Hígado/metabolismo , Ratones , Ratones Noqueados , Poli I-C , Reacción en Cadena de la Polimerasa , Proteínas Proto-Oncogénicas c-myc/genéticaRESUMEN
The development and homeostasis of the immune system requires an exquisite balance between cell proliferation and cell death. In this review, we discuss several in vivo and in vitro models that have been developed to help understand the importance of apoptosis during B and T cell development and activation.