Induction of Erythroid Differentiation in Human Erythroleukemia Cells by Depletion of Malic Enzyme 2

Malic enzyme 2 (ME2) is a mitochondrial enzyme that catalyzes the conversion of malate to pyruvate and CO2 and uses NAD as a cofactor. Higher expression of this enzyme correlates with the degree of cell de-differentiation. We found that ME2 is expressed in K562 erythroleukemia cells, in which a number of agents have been found to induce differentiation either along the erythroid or the myeloid lineage. We found that knockdown of ME2 led to diminished proliferation of tumor cells and increased apoptosis in vitro. These findings were accompanied by differentiation of K562 cells along the erythroid lineage, as confirmed by staining for glycophorin A and hemoglobin production. ME2 knockdown also totally abolished growth of K562 cells in nude mice. Increased ROS levels, likely reflecting increased mitochondrial production, and a decreased NADPH/NADP+ ratio were noted but use of a free radical scavenger to decrease inhibition of ROS levels did not reverse the differentiation or apoptotic phenotype, suggesting that ROS production is not causally involved in the resultant phenotype. As might be expected, depletion of ME2 induced an increase in the NAD+/NADH ratio and ATP levels fell significantly. Inhibition of the malate-aspartate shuttle was insufficient to induce K562 differentiation. We also examined several intracellular signaling pathways and expression of transcription factors and intermediate filament proteins whose expression is known to be modulated during erythroid differentiation in K562 cells. We found that silencing of ME2 leads to phospho-ERK1/2 inhibition, phospho-AKT activation, increased GATA-1 expression and diminished vimentin expression. Metabolomic analysis, conducted to gain insight into intermediary metabolic pathways that ME2 knockdown might affect, showed that ME2 depletion resulted in high orotate levels, suggesting potential impairment of pyrimidine metabolism. Collectively our data point to ME2 as a potentially novel metabolic target for leukemia therapy

Lower land-use emissions responsible for increased net land carbon sink during the slow warming period

The terrestrial carbon sink accelerated during 1998–2012, concurrently with the slow warming period, but the mechanisms behind this acceleration are unclear. Here we analyse recent changes in the net land carbon sink (NLS) and its driving factors, using atmospheric inversions and terrestrial carbon models. We show that the linear trend of NLS during 1998–2012 is about 0.17 ± 0.05 Pg C yr−2 , which is three times larger than during 1980–1998 (0.05 ± 0.05 Pg C yr−2). According to terrestrial carbon model simulations, the intensification of the NLS cannot be explained by CO2 fertilization or climate change alone. We therefore use a bookkeeping model to explore the contribution of changes in land-use emissions and find that decreasing land-use emissions are the dominant cause of the intensification of the NLS during the slow warming period. This reduction of land-use emissions is due to both decreased tropical forest area loss and increased afforestation in northern temperate regions. The estimate based on atmospheric inversions shows consistently reduced land-use emissions, whereas another bookkeeping model did not reproduce such changes, probably owing to missing the signal of reduced tropical deforestation. These results highlight the importance of better constraining emissions from land-use change to understand recent trends in land carbon sinks