RNA interference targeting survivin exerts antitumoral effects in vitro and in established glioma xenografts in vivo

Malignant glioma represents the most common primary adult brain tumor in Western industrialized countries. Despite aggressive treatment modalities, the median survival duration for patients with glioblastoma multiforme (GBM), the highest grade malignant glioma, has not improved significantly over past decades. One promising approach to deal with GBM is the inactivation of proteins essential for survival or progression of glioma cells by means of RNA interference (RNAi) techniques. A likely candidate for an RNAi therapy of gliomas is the inhibitor of apoptosis protein survivin. Survivin is involved in 2 main cellular processes–cell division and inhibition of apoptosis. We show here that stable RNAi of survivin induced polyploidy, apoptosis, and impaired proliferation of human U343-MG, U373-MG, H4, and U87-MG cells and of primary glioblastoma cells. Proteome profiler arrays using U373-MG cells identified a novel set of differentially expressed genes upon RNAi-mediated survivin knockdown. In particular, the death receptor TRAIL R2/DR5 was strongly upregulated in survivin-depleted glioma cells, inducing an enhanced cytotoxic response of allogeneic human NK cells. Moreover, an experimental in vivo therapy using polyethylenimine (PEI)/siRNA complexes for survivin knockdown efficiently blocked tumor growth of established subcutaneous U373-MG tumors and enhanced survival of NMRInu/nu mice orthopically transplanted with U87-MG cells. We conclude that survivin is functionally relevant in gliomas and that PEI-mediated exogenous delivery of siRNA targeting survivin is a promising strategy for glioblastoma therapy

Mutant IDH1 Differently Affects Redox State and Metabolism in Glial Cells of Normal and Tumor Origin

Contains fulltext : 215221.pdf (publisher's version ) (Open Access)IDH1(R132H) (isocitrate dehydrogenase 1) mutations play a key role in the development of low-grade gliomas. IDH1(wt) converts isocitrate to alpha-ketoglutarate while reducing nicotinamide adenine dinucleotide phosphate (NADP(+)), whereas IDH1(R132H) uses alpha-ketoglutarate and NADPH to generate the oncometabolite 2-hydroxyglutarate (2-HG). While the effects of 2-HG have been the subject of intense research, the 2-HG independent effects of IDH1(R132H) are still ambiguous. The present study demonstrates that IDH1(R132H) expression but not 2-HG alone leads to significantly decreased tricarboxylic acid (TCA) cycle metabolites, reduced proliferation, and enhanced sensitivity to irradiation in both glioblastoma cells and astrocytes in vitro. Glioblastoma cells, but not astrocytes, showed decreased NADPH and NAD(+) levels upon IDH1(R132H) transduction. However, in astrocytes IDH1(R132H) led to elevated expression of the NAD-synthesizing enzyme nicotinamide phosphoribosyltransferase (NAMPT). These effects were not 2-HG mediated. This suggests that IDH1(R132H) cells utilize NAD(+) to restore NADP pools, which only astrocytes could compensate via induction of NAMPT. We found that the expression of NAMPT is lower in patient-derived IDH1-mutant glioma cells and xenografts compared to IDH1-wildtype models. The Cancer Genome Atlas (TCGA) data analysis confirmed lower NAMPT expression in IDH1-mutant versus IDH1-wildtype gliomas. We show that the IDH1 mutation directly affects the energy homeostasis and redox state in a cell-type dependent manner. Targeting the impairments in metabolism and redox state might open up new avenues for treating IDH1-mutant gliomas

Physiological analysis of leaf senescence of two rice cultivars with different yield potential Análise fisiológica da senescência foliar de duas cultivares de arroz com diferentes potenciais de produtividade

The objective of this work was to evaluate the physiological changes that occur in different leaves during the early and late grain-filling stages of two rice genotypes (Oryza sativa subsp. indica , BRS Pelota cultivar, and O. sativa subsp. japonica , BRS Firmeza cultivar), which present differences in grain yield potential. The plants were cultivated in greenhouse. Pigment content, chlorophyll fluorescence, electron transport and oxygen evolution rate were determined in the grain-filling stage, from the first to the forth leaf (top to bottom). Pigment content, photochemical efficiency of photosystem II and electron transport decreased significantly according to the position of leaves in 'BRS Pelota'. The BRS Firmeza cultivar shows higher pigment content and higher activity of the photosynthetic apparatus in comparison to 'BRS Pelota' during the grain-filling stage.<br>O objetivo deste trabalho foi avaliar as mudanças fisiológicas que ocorrem em diferentes folhas durante o início e o final do estádio de enchimento de grãos em dois genótipos de arroz (Oryza sativa subsp. indica cultivar BRS Pelota e O. sativa subsp. japonica cultivar BRS Firmeza) que apresentam diferenças no potencial de produção de grãos. As plantas foram cultivadas em casa de vegetação. Os teores de pigmentos, a fluorescência da clorofila e a taxa de liberação de oxigênio foram determinados no estádio de enchimento de grão da primeira à quarta folha (do topo à base). O teor de pigmentos, a eficiência fotoquímica do fotossistema II e o transporte de elétrons decresceram significativamente de acordo com a posição das folhas na cultivar BRS Pelota. A cultivar BRS Firmeza apresentou maior teor de pigmentos e maior atividade do aparato fotossintético em comparação à 'BRS Pelota' durante o estádio de enchimento de grão