RESUMEN
BCR/ABL is the causative agent of chronic myelogenous leukemia (CML). Through structure/function analysis, several protein motifs have been determined to be important for the development of leukemogenesis. Tyrosine177 of BCR is a Grb2 binding site required for BCR/ABL-induced CML in mice. In the current study, we use a mouse bone marrow transduction/transplantation system to demonstrate that addition of oncogenic NRAS (NRASG12D) to a vector containing a BCR/ABL(Y177F) mutant "rescues" the CML phenotype rapidly and efficiently. To further narrow down the pathways downstream of RAS that are responsible for this rescue effect, we utilize well-characterized RAS effector loop mutants and determine that the RAL pathway is important for rapid induction of CML. Inhibition of this pathway by a dominant negative RAL is capable of delaying disease progression. Results from the present study support the notion of RAL inhibition as a potential therapy for BCR/ABL-induced CML.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Carcinogénesis/metabolismo , Leucemia Mielógena Crónica BCR-ABL Positiva/metabolismo , Proteínas de Unión al GTP ral/metabolismo , Factor de Intercambio de Guanina Nucleótido ral/metabolismo , Proteínas ras/metabolismo , Animales , Western Blotting , Trasplante de Médula Ósea , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Vectores Genéticos , Ratones , Células 3T3 NIH , RetroviridaeRESUMEN
The Vav family of Rho guanine nucleotide exchange factors is thought to orchestrate signaling events downstream of lymphocyte antigen receptors. Elucidation of Vav function has been obscured thus far by the expression of three highly related family members. We generated mice lacking all Vav family proteins and show that Vav-null mice produce no functional T or B cells and completely fail to mount both T-dependent and T-independent humoral responses. Whereas T cell development is blocked at an early stage in the thymus, immature B lineage cells accumulate in the periphery but arrest at a late "transitional" stage. Mechanistically, we show that the Vav family is crucial for both TCR and B cell receptor (BCR)-induced Ca2+ signaling and, surprisingly, is only required for mitogen-activated protein kinase (MAPK) activation in developing and mature T cells but not in B cells. Thus, the abundance of immature B cells generated in Vav-null mice may be due to intact Ras/MAPK signaling in this lineage. Although the expression of Vav1 alone is sufficient for normal lymphocyte development, our data also reveal lineage-specific roles for Vav2 and Vav3, with the first demonstration that Vav3 plays a critical compensatory function in T cells. Together, we define an essential role for the entire Vav protein family in lymphocyte development and activation and establish the limits of functional redundancy both within this family and between Vav and other Rho-guanine nucleotide exchange factors.
Asunto(s)
Linfocitos B/metabolismo , Proteínas de Ciclo Celular , Diferenciación Celular/fisiología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Proteínas Oncogénicas/genética , Proteínas Proto-Oncogénicas/genética , Linfocitos T/metabolismo , Animales , Linfocitos B/enzimología , Linfocitos B/inmunología , Calcio/metabolismo , Señalización del Calcio/fisiología , Factores de Intercambio de Guanina Nucleótido , Ratones , Proteínas Quinasas Activadas por Mitógenos/inmunología , Proteínas Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-vav , Transducción de Señal/fisiología , Linfocitos T/enzimologíaRESUMEN
Vav1 is a guanine nucleotide exchange factor essential in the development and function of alphabeta lineage T cells. Here we report that in contrast to profound effects on pre-TCR- or alphabeta TCR-dependent events in thymocyte development, Vav1 deficiency has no detectable effect on the development of gammadelta T cells. Strikingly, however, these gammadelta T cells are markedly deficient in signaling through the gammadelta TCR, as evidenced by a lack of proliferation and cytokine production in response to stimulation with anti-gammadelta TCR antibodies. We propose that Vav1 has a unique and non-redundant role in the initiation of signaling downstream of the gammadelta TCR in lymphocytes.