MYC sensitises cells to apoptosis by driving energetic demand
Joy Edwards-HicksHuizhong SuMaurizio MangoliniKubra K YonetenJimi WillsGiovanny Rodriguez BlancoChristine YoungKevin ChoHeather BarkerMorwenna MuirAnia Naila GuerrieriXuefeng LiRachel WhiteP. ManasterskiElena MandrouKaren L H WillsJingyu ChenEmily AbrahamKianoosh SateriBin‐Zhi QianPeter BankheadMark J. ArendsNoor GammohAlex von KriegsheimGary J. PattiAndrew H. SimsJuan Carlos AcostaValerie G. BruntonKamil R. KrancMaria A. ChristophorouErika L. PearceIngo RingshausenAndrew J. Finch
21
Citation
59
Reference
10
Related Paper
Citation Trend
Abstract:
Abstract The MYC oncogene is a potent driver of growth and proliferation but also sensitises cells to apoptosis, which limits its oncogenic potential. MYC induces several biosynthetic programmes and primary cells overexpressing MYC are highly sensitive to glutamine withdrawal suggesting that MYC-induced sensitisation to apoptosis may be due to imbalance of metabolic/energetic supply and demand. Here we show that MYC elevates global transcription and translation, even in the absence of glutamine, revealing metabolic demand without corresponding supply. Glutamine withdrawal from MRC-5 fibroblasts depletes key tricarboxylic acid (TCA) cycle metabolites and, in combination with MYC activation, leads to AMP accumulation and nucleotide catabolism indicative of energetic stress. Further analyses reveal that glutamine supports viability through TCA cycle energetics rather than asparagine biosynthesis and that TCA cycle inhibition confers tumour suppression on MYC-driven lymphoma in vivo. In summary, glutamine supports the viability of MYC-overexpressing cells through an energetic rather than a biosynthetic mechanism.Keywords:
Catabolism
The effects of glutamine and asparagine on CHO cell growth,metabolism and antibody expression in batch culture with cells being centrifuged were investigated. It was found that glutamine can't be replaced by asparagine simply,CHO-dhfr cells can't grow normally in adding separately experiments. When glutamine and asparagine concentration reached 4mmol / L meanwhile can support normal growth of CHO cells. As the total amount of glutamine and asparagine increased( 0 to 24mmol / L),ammonia concentration increased linearly, however,ammonia production had no relationship with proportion of glutamine and asparagine. Furthermore, improving the asparagine and glutamine proportion moderately not only can increase the antibody production but also decrease the lactate production. In brief,the foundation of supplement of glutamine and asparagine in development and optimization of mediums and efficient fed-batch process for antibody production was established.
Asparagine synthetase
Cite
Citations (0)
Chinese hamster ovary (CHO) cell lines are grown in cultures with varying asparagine and glutamine concentrations, but further study is needed to characterize the interplay between these amino acids. By following 13 C-glucose, 13 C-glutamine, and 13 C-asparagine tracers using metabolic flux analysis (MFA), CHO cell metabolism was characterized in an industrially relevant fed-batch process under glutamine supplemented and low glutamine conditions during early and late exponential growth. For both conditions MFA revealed glucose as the primary carbon source to the tricarboxylic acid (TCA) cycle followed by glutamine and asparagine as secondary sources. Early exponential phase CHO cells prefer glutamine over asparagine to support the TCA cycle under the glutamine supplemented condition, while asparagine was critical for TCA activity for the low glutamine condition. Overall TCA fluxes were similar for both conditions due to the trade-offs associated with reliance on glutamine and/or asparagine. However, glutamine supplementation increased fluxes to alanine, lactate and enrichment of glutathione, N-acetyl-glucosamine and pyrimidine-containing-molecules. The late exponential phase exhibited reduced central carbon metabolism dominated by glucose, while lactate reincorporation and aspartate uptake were preferred over glutamine and asparagine. These 13 C studies demonstrate that metabolic flux is process time dependent and can be modulated by varying feed composition.
Asparagine synthetase
Metabolic flux analysis
Cite
Citations (23)
Asparaginase
Asparagine synthetase
Cite
Citations (274)
Asparaginase
Asparagine synthetase
Essential amino acid
Cite
Citations (5)
This chapter contains sections titled: Introduction Nonenzymic Degradation of Glutamine, Asparagine, and Glutathione Distribution of Glutamine, Asparagine, and Glutathione Asparagine and Glutamine Metabolism in Microorganisms Amide Metabolism in Plants Asparagine and Glutamine Metabolism in Intact Animals Glutamine Metabolism and Fertilization Glutathione Metabolism in Mammals Synthesis of Glutamine, Asparagine, and Glutathione Enzymic Splitting of Asparagine, Glutamine, and Glutathione
Cite
Citations (64)
Chinese Hamster Ovary (CHO) cell lines are grown in cultures with varying asparagine and glutamine concentrations, but further study is needed to characterize the interplay between these amino acids. By following 13C-glucose, 13C-glutamine, and 13C-asparagine tracers using metabolic flux analysis (MFA), CHO cell metabolism was characterized in an industrially relevant fed-batch process under glutamine supplemented and low glutamine conditions during early and late exponential growth. For both conditions MFA revealed glucose as the primary carbon source to the tricarboxylic acid (TCA) cycle followed by glutamine and asparagine as secondary sources. Early exponential phase CHO cells prefer glutamine over asparagine to support the TCA cycle under the glutamine supplemented condition, while asparagine was critical for TCA activity for the low glutamine condition. Overall TCA fluxes were similar for both conditions due to the trade-offs associated with reliance on glutamine and/or asparagine. However, glutamine supplementation increased fluxes to alanine, lactate and enrichment of glutathione, N-Acetyl-Glucosamine (NAG) and pyrimidine-containing-molecules. The late exponential phase exhibited reduced central carbon metabolism dominated by glucose, while lactate reincorporation and aspartate uptake were preferred over glutamine and asparagine. These 13C studies demonstrate that metabolic flux is process time dependent and can be modulated by varying feed composition.
Metabolic flux analysis
Asparagine synthetase
Alanine
Cite
Citations (1)
Abstract Osteoblast differentiation is sequentially characterized by high rates of proliferation followed by increased protein and matrix synthesis, processes that require substantial amino acid acquisition and production. How osteoblasts obtain or maintain intracellular amino acid production is poorly understood. Here we identify Slc1a5 as a critical amino acid transporter during bone development. Using a genetic and metabolomic approach, we show Slc1a5 acts cell autonomously in osteoblasts to import glutamine and asparagine. Deleting Slc1a5 or reducing either glutamine or asparagine availability prevents protein synthesis and osteoblast differentiation. Mechanistically, glutamine and asparagine metabolism support amino acid biosynthesis. Thus, osteoblasts depend on Slc1a5 to provide glutamine and asparagine, which are subsequently used to produce non-essential amino acids and support osteoblast differentiation and bone development.
Asparagine synthetase
Cite
Citations (1)
Abstract Most proliferating cells consume exogenous glutamine to fuel biosynthesis and cell growth, which leads to its marked depletion from the microenvironment. Glutamine provides carbon and nitrogen for a wide array of biomolecules, among which are non-essential amino acids, glucosamine and nucleotides; in addition, carbon backbone of glutamine is used as a preferred anaplerotic input into the TCA cycle. However, as recent in vivo metabolomic studies indicate, a number of tumor types are capable of utilizing alternative anaplerotic routes and, in fact, synthesize glutamine de novo. In addition, select cancer cell lines are able to proliferate in the absence of exogenous glutamine in culture medium. However, the metabolic and regulatory underpinnings that enable the cellular adaptation to glutamine deficit and de novo glutamine production need further investigation. In this work, we identified the non-essential amino acid asparagine as essential for the survival and proliferation of cancer cells in the absence of exogenous glutamine supply. Our data indicate that asparagine, though structurally related to glutamine, is not catabolized in the same manner. Instead, asparagine acts by restoring protein translation, which becomes profoundly compromised in the absence of glutamine. This, in turn, enables the post-transcriptional adaptive upregulation of glutamine synthetase (GLUL), which is required for glutamine-independent growth. Taken together, these findings show that the cellular adaptation to glutamine-limiting conditions relies upon the availability of asparagine, which may represent an important therapeutic vulnerability. Citation Format: Natalya N. Pavlova, Ji Zhang, Craig B. Thompson. Asparagine drives translational adaptation of cancer cells to glutamine deficit. [abstract]. In: Proceedings of the AACR Special Conference on Translational Control of Cancer: A New Frontier in Cancer Biology and Therapy; 2016 Oct 27-30; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2017;77(6 Suppl):Abstract nr B39.
Asparagine synthetase
Cite
Citations (0)
Abstract One of the best characterized examples of metabolic changes occurring in a wide range of cancer cells is dependence on the non-essential amino acid glutamine. We identified requirements for several other non-essential amino acids, including asparagine, across a panel of glutamine-dependent and –independent breast and lung cancer cell lines. Glutamine and asparagine metabolism are tightly linked. Therefore, we hypothesized that concurrent targeting of the metabolism of these two non-essential amino acids could cooperatively interfere with tumor cell growth. Treatment with a glutaminase inhibitor, BPTES, and concurrent enzymatic depletion of asparagine produced a synergistic antiproliferative effect in many, but not all, cancer cell lines. Mechanistic data suggest that metabolic circuitry gets co-opted to maintain amino acid pools following combined targeting of glutamine and asparagine metabolism, resulting in mitochondrial dysfunction and redox imbalance. Following combination treatment, alterations in mitochondrial metabolism and cystine influx were observed, but overall amino acid levels remained surprisingly stable. These metabolic changes were reflected in cellular functional readouts as measured by Seahorse analysis and measurement of ROS levels. Importantly, they were also linked to the synergistic antiproliferative phenotype. Our results provide new mechanistic insight into the interplay between glutamine and asparagine metabolism in cancer cells and suggest that dual targeting of metabolically linked non-essential amino acids may be promising as a therapeutic strategy. Citation Format: Cicely L. Schramm, Zhi Xie, Erick Kindt, Wenlin Li, Fang Wang, Valeria R. Fantin. Concurrent targeting of glutamine and asparagine metabolism produces synergistic inhibition of tumor cell proliferation. [abstract]. In: Proceedings of the AACR Special Conference: Metabolism and Cancer; Jun 7-10, 2015; Bellevue, WA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(1_Suppl):Abstract nr A85.
Asparagine synthetase
Glutaminase
Cite
Citations (0)
Aspartic acid
Glutamic acid
Asparagine synthetase
Cite
Citations (12)