90 research outputs found
Effect of Monoethanolamine on the Lipid Metabolism of Prostate Cancer Cells
The ability of cancer cells to synthesize and store lipids in the form of lipid droplets (LDs) gives them a proliferative advantage over normal cells. Here, we elucidate the effect of monoethanolamine (Etn) on the lipid metabolism of LD-rich prostate carcinoma (PC3) cells. Etn is a ubiquitous molecule that serves as a phospholipid precursor and has been shown to have anticancer activity, although its mechanism of action is not fully understood. PC3 cells were treated with Etn and evaluated with a combination of biochemical and cell biology techniques. Treatment with Etn caused a decrease in cytosolic LD abundance induced by Ca2+-stimulated lipolysis. Our study effectively implicates the disruption of lipid metabolism as a contributor to Etn-induced cell death. This finding opens new avenues for the development of LD-targeted anticancer therapy
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Mitochondrial Inheritance and Function in the Lifespan Control of Budding Yeast
Mitochondria are essential organelles that cannot be synthesized de novo and must be inherited by daughter cells. During cell division, mitochondria align along the mother- daughter axis of the dividing cell, exhibit bidirectional poleward movement and are anchored at the cell poles. Mitochondria anchored at the bud tip and thus destined to be inherited by the daughter cell, show markers of increased fitness, lower superoxide burden and less oxidizing mitochondria, while less fit mitochondria are retained in the mother. In this work, the mechanism for anchorage of fit mitochondria to the bud tip and its effect on yeast lifespan determination are presented. Mitochondria at the bud tip are associated with cortical ER (cER) sheets underlying the plasma membrane. Mmr1p, a member of the DSL1 family of tethering proteins, mediates anchorage of mitochondria at the bud tip by binding to both mitochondria and cER at this site. A conserved protein phosphatase, Ptc1p, regulates mitochondrial anchorage by dephosphorylation of Mmr1p.
Mitochondrial fitness decreases as a function of age, yet retention of less fit mitochondria occurs to the same extent in young and older cells. Disruption of mitochondrial anchorage at the bud tip by deletion of MMR1 results in a severe lifespan anomaly, such that some cells have drastically reduced lifespan and markers of aged cells, while others show increased lifespan and markers of young cells. Loss of anchorage also leads to defects in mitochondrial quality control during inheritance and mitochondrial fitness correlates to the aging phenotypes observed in mmr1-delta cells. These findings support the model that the mitochondrial inheritance machinery promotes retention of lower-functioning mitochondria in mother cells and that this process contributes to both mother- daughter age asymmetry and age-associated declines in cellular fitness
The bioenergetic âCK Clampâ technique detects substrate-specific changes in mitochondrial respiration and membrane potential during early VML injury pathology
Volumetric muscle loss (VML) injuries are characterized by non-recoverable loss of tissue resulting in contractile and metabolic dysfunction. The characterization of metabolic dysfunction in volumetric muscle loss-injured muscle has been interpreted from permeabilized myofiber respiration experiments involving saturating ADP levels and non-physiologic ATP:ADP concentration ratios. The extent to which this testing condition obscures the analysis of mitochondrial (dys) function after volumetric muscle loss injury is unclear. An alternative approach is described that leverages the enzymatic reaction of creatine kinase and phosphocreatine to assess mitochondrial respiration and membrane potential at clamped physiologic ATP:ADP ratios, âCK Clamp.â The objective of this study was to validate the CK Clamp in volumetric muscle loss-injured muscle and to detect differences that may exist between volumetric muscle loss-injured and uninjured muscles at 1, 3, 5, 7, 10, and 14Â days post-injury. Volumetric muscle loss-injured muscle maintains bioenergetic features of the CK Clamp approach, i.e., mitochondrial respiration rate (JO2) titters down and mitochondrial membrane potential is more polarized with increasing ATP:ADP ratios. Pyruvate/malate/succinate-supported JO2 was significantly less in volumetric muscle loss-injured muscle at all timepoints compared to uninjured controls (â26% to â84%, p < 0.001) and electron conductance was less at day 1 (â60%), 5 (â52%), 7 (â35%), 10 (â59%), and 14 (â41%) (p < 0.001). Palmitoyl-carnitine/malate-supported JO2 and electron conductance were less affected following volumetric muscle loss injury. volumetric muscle loss-injury also corresponded with a more polarized mitochondrial membrane potential across the clamped ATP:ADP ratios at day 1 and 10 (pyruvate and palmitoyl-carnitine, respectively) (+5%, p < 0.001). This study supports previous characterizations of metabolic dysfunction and validates the CK Clamp as a tool to investigate bioenergetics in traumatically-injured muscle
Thyroid hormone regulates distinct paths to maturation in pigment cell lineages
Thyroid hormone (TH) regulates diverse developmental events and can drive disparate cellular outcomes. In zebrafish, TH has opposite effects on neural crest derived pigment cells of the adult stripe pattern, limiting melanophore population expansion, yet increasing yellow/orange xanthophore numbers. To learn how TH elicits seemingly opposite responses in cells having a common embryological origin, we analyzed individual transcriptomes from thousands of neural crest-derived cells, reconstructed developmental trajectories, identified pigment cell-lineage specific responses to TH, and assessed roles for TH receptors. We show that TH promotes maturation of both cell types but in distinct ways. In melanophores, TH drives terminal differentiation, limiting final cell numbers. In xanthophores, TH promotes accumulation of orange carotenoids, making the cells visible. TH receptors act primarily to repress these programs when TH is limiting. Our findings show how a single endocrine factor integrates very different cellular activities during the generation of adult form
Neural G0:a quiescent-like state found in neuroepithelial-derived cells and glioma
Singleâcell RNA sequencing has emerged as a powerful tool for resolving cellular states associated with normal and maligned developmental processes. Here, we used scRNAâseq to examine the cell cycle states of expanding human neural stem cells (hNSCs). From these data, we constructed a cell cycle classifier that identifies traditional cell cycle phases and a putative quiescentâlike state in neuroepithelialâderived cell types during mammalian neurogenesis and in gliomas. The Neural G0 markers are enriched with quiescent NSC genes and other neurodevelopmental markers found in nonâdividing neural progenitors. Putative glioblastoma stemâlike cells were significantly enriched in the Neural G0 cell population. Neural G0 cell populations and gene expression are significantly associated with less aggressive tumors and extended patient survival for gliomas. Genetic screens to identify modulators of Neural G0 revealed that knockout of genes associated with the Hippo/Yap and p53 pathways diminished Neural G0 in vitro, resulting in faster G1 transit, downâregulation of quiescenceâassociated markers, and loss of Neural G0 gene expression. Thus, Neural G0 represents a dynamic quiescentâlike state found in neuroepithelialâderived cells and gliomas
Telomerase inhibition is an effective therapeutic strategy in TERT promoter mutant-glioblastoma models with low tumor volume
Background
Glioblastoma is one of the most lethal forms of cancer, with 5-year survival rates of only 6%. Glioblastoma-targeted therapeutics have been challenging to develop due to significant inter- and intra-tumoral heterogeneity. Telomerase reverse transcriptase gene (TERT) promoter mutations are the most common known clonal oncogenic mutations in glioblastoma. Telomerase is therefore considered to be a promising therapeutic target against this tumor. However, an important limitation of this strategy is that cell death does not occur immediately after telomerase ablation, but rather after several cell divisions required to reach critically short telomeres. We, therefore, hypothesize that telomerase inhibition would only be effective in glioblastomas with low tumor burden.
Methods
We used CRISPR interference to knock down TERT expression in TERT promoter-mutant glioblastoma cell lines and patient-derived models. We then measured viability using serial proliferation assays. We also assessed for features of telomere crisis by measuring telomere length and chromatin bridge formation. Finally, we used a doxycycline-inducible CRISPR interference system to knock down TERT expression in vivo early and late in tumor development.
Results
Upon TERT inactivation, glioblastoma cells lose their proliferative ability over time and exhibit telomere shortening and chromatin bridge formation. In vivo, survival is only prolonged when TERT knockdown is induced shortly after tumor implantation, but not when the tumor burden is high.
Conclusions
Our results support the idea that telomerase inhibition would be most effective at treating glioblastomas with low tumor burden, for example in the adjuvant setting after surgical debulking and chemoradiation
Topoisomerase II\u3b2 mediates the resistance of glioblastoma stem cells to replication stress-inducing drugs
The mesenchymal state in cancer is usually associated with poor prognosis due to the metastatic predisposition and the hyper-activated metabolism. Exploiting cell glucose metabolism we propose a new method to detect mesenchymal-like cancer cells. We demonstrate that the uptake of glucose-coated magnetic nanoparticles (MNPs) by mesenchymal-like cells remains constant when the glucose in the medium is increased from low (5.5 mM) to high (25 mM) concentration, while the MNPs uptake by epithelial-like cells is significantly reduced. These findings reveal that the glucose-shell of MNPs plays a major role in recognition of cells with high-metabolic activity. By selectively blocking the glucose transporter 1 channels we showed its involvement in the internalization process of glucose-coated MNPs. Our results suggest that glucose-coated MNPs can be used for metabolic-based assays aimed at detecting cancer cells and that can be used to selectively target cancer cells taking advantage, for instance, of the magnetic-thermotherapy
Generational distribution of a Candida glabrata population: Resilient old cells prevail, while younger cells dominate in the vulnerable host.
Similar to other yeasts, the human pathogen Candida glabrata ages when it undergoes asymmetric, finite cell divisions, which determines its replicative lifespan. We sought to investigate if and how aging changes resilience of C. glabrata populations in the host environment. Our data demonstrate that old C. glabrata are more resistant to hydrogen peroxide and neutrophil killing, whereas young cells adhere better to epithelial cell layers. Consequently, virulence of old compared to younger C. glabrata cells is enhanced in the Galleria mellonella infection model. Electron microscopy images of old C. glabrata cells indicate a marked increase in cell wall thickness. Comparison of transcriptomes of old and young C. glabrata cells reveals differential regulation of ergosterol and Hog pathway associated genes as well as adhesion proteins, and suggests that aging is accompanied by remodeling of the fungal cell wall. Biochemical analysis supports this conclusion as older cells exhibit a qualitatively different lipid composition, leading to the observed increased emergence of fluconazole resistance when grown in the presence of fluconazole selection pressure. Older C. glabrata cells accumulate during murine and human infection, which is statistically unlikely without very strong selection. Therefore, we tested the hypothesis that neutrophils constitute the predominant selection pressure in vivo. When we altered experimentally the selection pressure by antibody-mediated removal of neutrophils, we observed a significantly younger pathogen population in mice. Mathematical modeling confirmed that differential selection of older cells is sufficient to cause the observed demographic shift in the fungal population. Hence our data support the concept that pathogenesis is affected by the generational age distribution of the infecting C. glabrata population in a host. We conclude that replicative aging constitutes an emerging trait, which is selected by the host and may even play an unanticipated role in the transition from a commensal to a pathogen state.post-print10768 K
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