Profiling cancer.

In the past couple of years, several very exciting studies have demonstrated the enormous power of gene-expression profiling for cancer classification and prediction of patient survival. In addition to promising a more accurate classification of cancer and therefore better treatment of patients, gene-expression profiling can result in the identification of novel potential targets for cancer therapy and a better understanding of the molecular mechanisms leading to cancer.

E2F target genes: unraveling the biology.

The E2F transcription factors are downstream effectors of the retinoblastoma protein (pRB) pathway and are required for the timely regulation of numerous genes essential for DNA replication and cell cycle progression. Several laboratories have used genome-wide approaches to discover novel target genes of E2F, leading to the identification of several hundred such genes that are involved not only in DNA replication and cell cycle progression, but also in DNA damage repair, apoptosis, differentiation and development. These new findings greatly enrich our understanding of how E2F controls transcription and cellular homeostasis.

Combination of Hypoglycemia and Metformin Impairs Tumor Metabolic Plasticity and Growth by Modulating the PP2A-GSK3ß-MCL-1 Axis.

Tumor cells may adapt to metabolic challenges by alternating between glycolysis and oxidative phosphorylation (OXPHOS). To target this metabolic plasticity, we combined intermittent fasting, a clinically feasible approach to reduce glucose availability, with the OXPHOS inhibitor metformin. In mice exposed to 24-h feeding/fasting cycles, metformin impaired tumor growth only when administered during fasting-induced hypoglycemia. Synergistic anti-neoplastic effects of the metformin/hypoglycemia combination were mediated by glycogen synthase kinase 3ß (GSK3ß) activation downstream of PP2A, leading to a decline in the pro-survival protein MCL-1, and cell death. Mechanistically, specific activation of the PP2A-GSK3ß axis was the sum of metformin-induced inhibition of CIP2A, a PP2A suppressor, and of upregulation of the PP2A regulatory subunit B56δ by low glucose, leading to an active PP2A-B56δ complex with high affinity toward GSK3ß.

Involvement of mitogen-activated protein kinase in the cytokine-regulated phosphorylation of transcription factor GATA-1.

Gene-targeting experiments in transgenic mice have revealed an essential role for GATA-1 in the normal differentiation and development of erythroid cells. GATA-1 is phosphorylated in vivo on seven of its serine residues; the regulation and function of GATA-1 phosphorylation, however, is not understood. Here we demonstrate a role for MAP kinase (MAPK) signalling in the control of GATA-1 phosphorylation. We show that EGF-induced MAPK signalling results in the phosphorylation of ectopically expressed GATA-1 in COS cells. This phosphorylation can be positively or negatively regulated by genetic manipulation of the MAPK pathway through expression of constitutively activated, or dominant-negative, mutants of MAPK kinase (MAPKK), an upstream regulator of MAPK activity. In vitro phosphorylation experiments using purified MAPK and either recombinant GATA-1 or synthetic GATA-1 peptides suggest that GATA-1 is a MAPK substrate with MAPK phosphorylation occurring primarily on Ser26 and Ser178. We also show that GATA-1 is phosphorylated in factor-dependent haemopoietic progenitor cells in response to cytokine-induced signalling. Through the further use of a dominant-negative MAPKK mutant as well as chemical inhibitors of specific MAPKs, we identify ERK as an in vivo GATA-1 kinase. Finally, we demonstrate that mutation of serines 26 and 178 compromises the ability of GATA-1 to interact with the LIM-only protein LMO2 when both proteins are expressed in COS cells. These data implicate receptor-mediated signalling through the MAPK pathway as a control point in the regulation of transcription factor GATA-1.

Beclin 1 restrains tumorigenesis through Mcl-1 destabilization in an autophagy-independent reciprocal manner.

Mcl-1 is a unique Bcl-2 family member that plays crucial roles in apoptosis. Apoptosis-unrelated functions of Mcl-1 are however emerging, further justifying its tight regulation. Here we unravel a novel mechanism of Mcl-1 regulation mediated by the haplo-insufficient tumour suppressor Beclin 1. Beclin 1 negatively modulates Mcl-1 stability in a reciprocal manner whereby depletion of one leads to the stabilization of the other. This co-regulation is independent of autophagy and of their physical interaction. Both Beclin 1 and Mcl-1 are deubiquitinated and thus stabilized by binding to a common deubiquitinase, USP9X. Beclin 1 and Mcl-1 negatively modulate the proteasomal degradation of each other through competitive displacement of USP9X. The analysis of patient-derived melanoma cells and tissue samples shows that the levels of Beclin 1 decrease, while Mcl-1 levels subsequently increase during melanoma progression in a significant inter-dependent manner. The identified inverse co-regulation of Beclin 1 and Mcl-1 represents a mechanism of functional counteraction in cancer.

Encefalopatía posterior reversible en preeclampsia / Reversible posterior encephalopathy in preeclampsia / Encefalopatia posterior reversível em pré-eclâmpsia

Resumen: El síndrome de encefalopatía posterior reversible (PRES por sus siglas en inglés Reversible Posterior Leukoencephalopathy Syndrome) es una entidad clínico-radiológica con datos claros de afección del lóbulo posterior por imagen por resonancia magnética. Fue descrito por primera vez en 1996 por Hinchey, se caracteriza clínicamente por cefalea, afección laberíntica, así como ceguera cortical, entre otros síntomas. Una de las poblaciones en riesgo de presentar esta complicación neurológica son las mujeres que padecen preeclampsia/eclampsia. El objetivo de este trabajo es exponer un caso de encefalopatía posterior reversible en una paciente con preeclampsia.

Abstract: Posterior reversible encephalopathy syndrome is a clinical-radiological entity, with clear data of vasogenic edema at the posterior lobe by magnetic resonance image. As First described in 1996 by Hinchey, clinical features are headache, labyrinthine affection, and cortical blindness among others symptoms preeclampsia/eclampsia among obstetric patients has been characterized for presenting as a risk factor for this neurologic complication. The aim of this paper is to present a clinical case of posterior reversible encephalopathy in a patient with pre-eclampsia.

Resumo: A síndrome da encefalopatia posterior reversível (PRES, sigla em Inglês) é uma entidade clínica radiológica, com clara evidência da afetação do lobo posterior por IRM.1 Descrita pela primeira vez em 1996 por Hinchey; clinicamente caracterizada por cefaleia, doença labiríntica e cegueira cortical entre outros sintomas. Uma das populações em risco de apresentar essa complicação neurológica são mulheres com pré-eclâmpsia/eclâmpsia. O objetivo deste trabalho é apresentar um caso de encefalopatia posterior reversível em uma paciente com pré-eclâmpsia.

ATAD2 is a novel cofactor for MYC, overexpressed and amplified in aggressive tumors.

The E2F and MYC transcription factors are critical regulators of cell proliferation and contribute to the development of human cancers. Here, we report on the identification of a novel E2F target gene, ATAD2, the predicted protein product of which contains both a bromodomain and an ATPase domain. The pRB-E2F pathway regulates ATAD2 expression, which is limiting for the entry into the S phase of the cell cycle. We show that ATAD2 binds the MYC oncogene and stimulates its transcriptional activity. ATAD2 maps to chromosome 8q24, 4.3 Mb distal to MYC, in a region that is frequently found amplified in cancer. Consistent with this, we show that ATAD2 expression is high in several human tumors and that the expression levels correlate with clinical outcome of breast cancer patients. We suggest that ATAD2 links the E2F and MYC pathways and contributes to the development of aggressive cancer through the enhancement of MYC-dependent transcription.

Acetylation and MAPK phosphorylation cooperate to regulate the degradation of active GATA-1.

Regulation of transcription requires mechanisms to both activate and terminate transcription factor activity. GATA-1 is a key haemopoietic transcription factor whose activity is increased by acetylation. We show here that acetylated GATA-1 is targeted for degradation via the ubiquitin/proteasome pathway. Acetylation positively signals ubiquitination, suggesting that activation by acetylation simultaneously marks GATA-1 for degradation. Promoter-specific MAPK phosphorylation then cooperates with acetylation to execute protein loss. The requirement for both modifications is novel and suggests a way by which degradation of the active protein can be specifically regulated in response to external phosphorylation-mediated signalling. As many transcription factors are activated by acetylation, we suggest that this might be a general mechanism to control transcription factor activity.

Induction of globin mRNA expression by interleukin-3 in a stem cell factor-dependent SV-40 T-antigen-immortalized multipotent hematopoietic cell line.

Erythropoiesis requires the stepwise action on immature progenitors of several growth factors, including stem cell factor (SCF), interleukin 3 (IL-3), and erythropoietin (Epo). Epo is required to sustain proliferation and survival of committed progenitors and might further modulate the level of expression of several erythroid genes, including globin genes. Here we report a new SCF-dependent immortalized mouse progenitor cell line (GATA-1 ts SCF) that can also grow in either Epo or IL-3 as the sole growth factor. When grown in SCF, these cells show an "open" chromatin structure of the beta-globin LCR, but do not significantly express globin. However, Epo or IL-3 induce globin expression and are required for its maintainance. This effect of IL-3 is unexpected as IL-3 was previously reported either to be unable to induce hemoglobinization, or even to antagonize it. This suggests that GATA-1 ts SCF cells may have progressed to a stage in which globin genes are already poised for expression and only require signal(s) that can be elicited by either Epo or IL-3. Through the use of inhibitors, we suggest that p38 may be one of the molecules modulating induction and maintenance of globin expression.