scholarly journals Quantitative analysis of amino acid metabolism in liver cancer links glutamate excretion to nucleotide synthesis

2020 ◽  
Vol 117 (19) ◽  
pp. 10294-10304 ◽  
Author(s):  
Avlant Nilsson ◽  
Jurgen R. Haanstra ◽  
Martin Engqvist ◽  
Albert Gerding ◽  
Barbara M. Bakker ◽  
...  
Keyword(s):  
Liver Cancer ◽  
Drug Targets ◽  
Tricarboxylic Acid Cycle ◽  
Cancer Cell Line ◽  
Glutamine Metabolism ◽  
Integrative Approach ◽  
Synthesis Rate ◽  
Nucleotide Synthesis ◽  
Depth Analysis ◽  
Glutamate Excretion

Many cancer cells consume glutamine at high rates; counterintuitively, they simultaneously excrete glutamate, the first intermediate in glutamine metabolism. Glutamine consumption has been linked to replenishment of tricarboxylic acid cycle (TCA) intermediates and synthesis of adenosine triphosphate (ATP), but the reason for glutamate excretion is unclear. Here, we dynamically profile the uptake and excretion fluxes of a liver cancer cell line (HepG2) and use genome-scale metabolic modeling for in-depth analysis. We find that up to 30% of the glutamine is metabolized in the cytosol, primarily for nucleotide synthesis, producing cytosolic glutamate. We hypothesize that excreting glutamate helps the cell to increase the nucleotide synthesis rate to sustain growth. Indeed, we show experimentally that partial inhibition of glutamate excretion reduces cell growth. Our integrative approach thus links glutamine addiction to glutamate excretion in cancer and points toward potential drug targets.

Mycobacterial DNA Replication as a Target for Antituberculosis Drug Discovery

2017 ◽  
Vol 17 (19) ◽  
pp. 2129-2142 ◽  
Author(s):  
Renata Płocinska ◽  
Malgorzata Korycka-Machala ◽  
Przemyslaw Plocinski ◽  
Jaroslaw Dziadek
Keyword(s):  
Drug Resistance ◽  
Dna Replication ◽  
Drug Targets ◽  
Rapid Development ◽  
New Drugs ◽  
Drug Resistant ◽  
Antituberculosis Drug ◽  
Nucleotide Synthesis ◽  
Antituberculosis Therapy ◽  
Optimal Drug

Background: Mycobacterium tuberculosis (M. tuberculosis), the causative agent of tuberculosis, is a leading infectious disease organism, causing millions of deaths each year. This serious pathogen has been greatly spread worldwide and recent years have observed an increase in the number of multi-drug resistant and totally drug resistant M. tuberculosis strains (WHO report, 2014). The danger of tuberculosis becoming an incurable disease has emphasized the need for the discovery of a new generation of antimicrobial agents. The development of novel alternative medical strategies, new drugs and the search for optimal drug targets are top priority areas of tuberculosis research. Factors: Key characteristics of mycobacteria include: slow growth, the ability to transform into a metabolically silent - latent state, intrinsic drug resistance and the relatively rapid development of acquired drug resistance. These factors make finding an ideal antituberculosis drug enormously challenging, even if it is designed to treat drug sensitive tuberculosis strains. A vast majority of canonical antibiotics including antituberculosis agents target bacterial cell wall biosynthesis or DNA/RNA processing. Novel therapeutic approaches are being tested to target mycobacterial cell division, twocomponent regulatory factors, lipid synthesis and the transition between the latent and actively growing states. Discussion and Conclusion: This review discusses the choice of cellular targets for an antituberculosis therapy, describes putative drug targets evaluated in the recent literature and summarizes potential candidates under clinical and pre-clinical development. We focus on the key cellular process of DNA replication, as a prominent target for future antituberculosis therapy. We describe two main pathways: the biosynthesis of nucleic acids precursors – the nucleotides, and the synthesis of DNA molecules. We summarize data regarding replication associated proteins that are critical for nucleotide synthesis, initiation, unwinding and elongation of the DNA during the replication process. They are pivotal processes required for successful multiplication of the bacterial cells and hence they are extensively investigated for the development of antituberculosis drugs. Finally, we summarize the most potent inhibitors of DNA synthesis and provide an up to date report on their status in the clinical trials.

scholarly journals Cytotoxic Profiling of Annotated and Diverse Chemical Libraries Using Quantitative High-Throughput Screening

2019 ◽  
Vol 25 (1) ◽  
pp. 9-20 ◽  
Author(s):  
Olivia W. Lee ◽  
Shelley Austin ◽  
Madison Gamma ◽  
Dorian M. Cheff ◽  
Tobie D. Lee ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cancer Cell ◽  
High Throughput ◽  
High Throughput Screening ◽  
Drug Targets ◽  
Large Scale ◽  
Early Stage ◽  
Cancer Cell Line ◽  
Hek 293 ◽  
Cytotoxic Compounds

Cell-based phenotypic screening is a commonly used approach to discover biological pathways, novel drug targets, chemical probes, and high-quality hit-to-lead molecules. Many hits identified from high-throughput screening campaigns are ruled out through a series of follow-up potency, selectivity/specificity, and cytotoxicity assays. Prioritization of molecules with little or no cytotoxicity for downstream evaluation can influence the future direction of projects, so cytotoxicity profiling of screening libraries at an early stage is essential for increasing the likelihood of candidate success. In this study, we assessed the cell-based cytotoxicity of nearly 10,000 compounds in the National Institutes of Health, National Center for Advancing Translational Sciences annotated libraries and more than 100,000 compounds in a diversity library against four normal cell lines (HEK 293, NIH 3T3, CRL-7250, and HaCat) and one cancer cell line (KB 3-1, a HeLa subline). This large-scale library profiling was analyzed for overall screening outcomes, hit rates, pan-activity, and selectivity. For the annotated library, we also examined the primary targets and mechanistic pathways regularly associated with cell death. To our knowledge, this is the first study to use high-throughput screening to profile a large screening collection (>100,000 compounds) for cytotoxicity in both normal and cancer cell lines. The results generated here constitute a valuable resource for the scientific community and provide insight into the extent of cytotoxic compounds in screening libraries, allowing for the identification and avoidance of compounds with cytotoxicity during high-throughput screening campaigns.

scholarly journals In vivo 13C MRS in the mouse brain at 14.1 Tesla and metabolic flux quantification under infusion of [1,6-13C2]glucose

2017 ◽  
Vol 38 (10) ◽  
pp. 1701-1714 ◽  
Author(s):  
Marta Lai ◽  
Bernard Lanz ◽  
Carole Poitry-Yamate ◽  
Jackeline F Romero ◽  
Corina M Berset ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Metabolic Flux ◽  
Direct Detection ◽  
Tricarboxylic Acid Cycle ◽  
Quantitative Information ◽  
Glutamine Metabolism ◽  
Resonance Spectroscopy ◽  
Flux Analysis ◽  
Carboxylase Activity

In vivo 13C magnetic resonance spectroscopy (MRS) enables the investigation of cerebral metabolic compartmentation while, e.g. infusing 13C-labeled glucose. Metabolic flux analysis of 13C turnover previously yielded quantitative information of glutamate and glutamine metabolism in humans and rats, while the application to in vivo mouse brain remains exceedingly challenging. In the present study, 13C direct detection at 14.1 T provided highly resolved in vivo spectra of the mouse brain while infusing [1,6-13C2]glucose for up to 5 h. 13C incorporation to glutamate and glutamine C4, C3, and C2 and aspartate C3 were detected dynamically and fitted to a two-compartment model: flux estimation of neuron-glial metabolism included tricarboxylic acid cycle (TCA) flux in astrocytes (Vg = 0.16 ± 0.03 µmol/g/min) and neurons (VTCAn = 0.56 ± 0.03 µmol/g/min), pyruvate carboxylase activity (VPC = 0.041 ± 0.003 µmol/g/min) and neurotransmission rate (VNT = 0.084 ± 0.008 µmol/g/min), resulting in a cerebral metabolic rate of glucose (CMRglc) of 0.38 ± 0.02 µmol/g/min, in excellent agreement with that determined with concomitant 18F-fluorodeoxyglucose positron emission tomography (18FDG PET).We conclude that modeling of neuron-glial metabolism in vivo is accessible in the mouse brain from 13C direct detection with an unprecedented spatial resolution under [1,6-13C2]glucose infusion.

scholarly journals Synergetic Effect of SLN-Curcumin and LDH-5-Fu on SMMC-7721 Liver Cancer Cell Line

2013 ◽  
Vol 28 (8) ◽  
pp. 579-587 ◽  
Author(s):  
Rongrong Zhu ◽  
Xianzheng Wu ◽  
Yu Xiao ◽  
Bo Gao ◽  
Qian Xie ◽  
...  
Keyword(s):  
Liver Cancer ◽  
Cell Line ◽  
Cancer Cell ◽  
Synergetic Effect ◽  
Cancer Cell Line ◽  
Liver Cancer Cell ◽  
Liver Cancer Cell Line

scholarly journals EVALUATION OF TOTAL PHENOLS, TOTAL FLAVONOIDS, ANTIOXIDANT, AND ANTICANCER ACTIVITY OF MUCUNA PRURIENS SEED

2018 ◽  
Vol 11 (3) ◽  
pp. 242
Author(s):  
Kavitha K
Keyword(s):  
Liver Cancer ◽  
Anticancer Activity ◽  
Cell Line ◽  
Cancer Cell Line ◽  
Ethanol Extract ◽  
Biological Properties ◽  
Seed Extract ◽  
Mucuna Pruriens ◽  
Total Flavonoids ◽  
Total Phenols

 Objective: In recent years, herbal plants have been got more attention due to their diverse presence of phytochemicals and its biological properties. Hepatocellular carcinoma is one of the major worldwide problems primarily caused by hepatitis B and C virus infection. End-stage liver cancer treatment options are limited, thus requiring expensive liver transplantation which is not available in many countries.Methods: In the present study, the Mucuna pruriens seed extract was analyzed for phytochemicals, antioxidant activity, total phenols, and total flavonoids content. The seed extract was further analyzed for its anticancer activity by culturing liver cancer cell line. The above protocols were done by standard methods.Results: The seed extract of M. pruriens revealed more number of phytochemicals in different organic solvents. 1,1- Diphenyl- 2- picrylhydrozyl scavenging activity of plant extract was more in ethanol extract (98.051±0.547) among all other solvents. The total phenols and flavonoids content in ethanol extract were 46.442±0.353 mg gallic acid equivalent/g and 2.254±0.647 mg - quercetin equivalent /g, respectively. IC50 value of 3-(4, 5-dimethythiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) analysis of Hep-2 cell line was 150 (μg/ml).Conclusion: The present study revealed about the phytochemical contents and antioxidant potential of M. pruriens seeds. Further, the MTT analysis proved that the seed extract was effective against cancer cells and also used to treat many diseases.

scholarly journals Anabolic SIRT4 Exerts Retrograde Control over TORC1 Signaling by Glutamine Sparing in the Mitochondria

2019 ◽  
Vol 40 (2) ◽  
Author(s):  
Eisha Shaw ◽  
Manasi Talwadekar ◽  
Zeenat Rashida ◽  
Nitya Mohan ◽  
Aishwarya Acharya ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Tricarboxylic Acid Cycle ◽  
Tricarboxylic Acid ◽  
Glutamine Metabolism ◽  
Peroxisome Proliferator ◽  
Nutrient Inputs ◽  
Peroxisome Proliferator Activated Receptor ◽  
Fed State ◽  
Acid Cycle ◽  
Mitochondrial Functions

ABSTRACT Anabolic and catabolic signaling mediated via mTOR and AMPK (AMP-activated kinase) have to be intrinsically coupled to mitochondrial functions for maintaining homeostasis and mitigate cellular/organismal stress. Although glutamine is known to activate mTOR, whether and how differential mitochondrial utilization of glutamine impinges on mTOR signaling has been less explored. Mitochondrial SIRT4, which unlike other sirtuins is induced in a fed state, is known to inhibit catabolic signaling/pathways through the AMPK-PGC1α/SIRT1–peroxisome proliferator-activated receptor α (PPARα) axis and negatively regulate glutamine metabolism via the tricarboxylic acid cycle. However, physiological significance of SIRT4 functions during a fed state is still unknown. Here, we establish SIRT4 as key anabolic factor that activates TORC1 signaling and regulates lipogenesis, autophagy, and cell proliferation. Mechanistically, we demonstrate that the ability of SIRT4 to inhibit anaplerotic conversion of glutamine to α-ketoglutarate potentiates TORC1. Interestingly, we also show that mitochondrial glutamine sparing or utilization is critical for differentially regulating TORC1 under fed and fasted conditions. Moreover, we conclusively show that differential expression of SIRT4 during fed and fasted states is vital for coupling mitochondrial energetics and glutamine utilization with anabolic pathways. These significant findings also illustrate that SIRT4 integrates nutrient inputs with mitochondrial retrograde signals to maintain a balance between anabolic and catabolic pathways.

scholarly journals Secondary Metabolite Mapping Identifies Peony Episperm Inhibitors of Human Hepatoma Cells

2019 ◽  
Vol 14 (7) ◽  
pp. 1934578X1986031
Author(s):  
Xiao Yang ◽  
Ruixue Deng ◽  
Pu Liu ◽  
Jiangxia Hu ◽  
Wentian Niu ◽  
...  
Keyword(s):  
Liver Cancer ◽  
Secondary Metabolite ◽  
Principal Components ◽  
Human Liver ◽  
Cancer Cell Line ◽  
Least Squares Regression ◽  
Human Hepatoma Cells ◽  
Liquid Chromatograph ◽  
Medicinal Uses ◽  
Human Liver Cancer

As the main by-product during the oil production of peony seeds, the episperm is traditionally used as a lead component in folk herbal formulas for the cancer treatment in China. However, the investigation of its phytochemical foundation underlying anticancer effects remains an ongoing challenge. The work therefore determined growth inhibition activities of 8 solvent extracts of peony episperms in the human liver cancer cell line. This activity was then mapped onto the secondary metabolite profile of extracts by principal components analysis (PCA). The top 3 principal components of High Performance Liquid Chromatograph (HPLC)-PCA map discriminated extract activities mainly based on the differential content of 5 stilbene compounds, which were then tested individually. The trans-ε-viniferin, gnetin-H, suffruticosol A, suffruticosol B, and suffruticosol C were thus determined as growth inhibitors and apoptosis inducers of human liver cancer cells with activities comparable to that of the antineoplastic cisplatin. A partial least squares regression-HPLC model was also constructed for the prediction of inhibitory effects of peony episperm extracts. These results expand the fundamental understanding of the peony episperms and support its current medicinal uses in China. Moreover, PCA-mediated secondary metabolite mapping was proved to be an efficient approach to qualify biomedical products required for pharmaceutical and medicinal uses.

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