Inhibition of nucleotide synthesis promotes replicative senescence of human mammary epithelial cells

Delfarah, Alireza; Parrish, Sydney; Jm, Yang; Seo, Frances; Li, Si; Wang, Pin; Graham, Nicholas A.

doi:10.1101/423665

Cited by 4 publications

(4 citation statements)

References 63 publications

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“…In line with previous results [65], we observed decreased uridine levels in the ageing spleen, suggesting uridine and taurine as robust spleen-specific biomarker candidates for the ageing process. Accordingly, reduced levels of uridine might be associated with increased cellular senescence in all tissues, as uridine has been shown to affect senescence in human mammary epithelial cells [66]. During ageing, senescent cells reduce production of nucleotides, the essential building blocks of DNA, implying that precursors like inosine, uridine, uracil and nicotinamide might also be found at abnormal levels [66].…”

Section: Discussionmentioning

confidence: 99%

“…Accordingly, reduced levels of uridine might be associated with increased cellular senescence in all tissues, as uridine has been shown to affect senescence in human mammary epithelial cells [66]. During ageing, senescent cells reduce production of nucleotides, the essential building blocks of DNA, implying that precursors like inosine, uridine, uracil and nicotinamide might also be found at abnormal levels [66]. Taken together, decreased uridine concentrations in the brain, heart, kidney, liver and spleen of aged mice indicate that uridine may be a general biomarker for ageing.…”

Section: Discussionmentioning

confidence: 99%

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Tissue-Specific Landscape of Metabolic Dysregulation during Ageing

et al. 2021

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The dysregulation of cellular metabolism is a hallmark of ageing. To understand the metabolic changes that occur as a consequence of the ageing process and to find biomarkers for age-related diseases, we conducted metabolomic analyses of the brain, heart, kidney, liver, lung and spleen in young (9–10 weeks) and old (96–104 weeks) wild-type mice [mixed genetic background of 129/J and C57BL/6] using NMR spectroscopy. We found differences in the metabolic fingerprints of all tissues and distinguished several metabolites to be altered in most tissues, suggesting that they may be universal biomarkers of ageing. In addition, we found distinct tissue-clustered sets of metabolites throughout the organism. The associated metabolic changes may reveal novel therapeutic targets for the treatment of ageing and age-related diseases. Moreover, the identified metabolite biomarkers could provide a sensitive molecular read-out to determine the age of biologic tissues and organs and to validate the effectiveness and potential off-target effects of senolytic drug candidates on both a systemic and tissue-specific level.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Tissue-Specific Landscape of Metabolic Dysregulation during Ageing

et al. 2021

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“…Recent metabolomic approaches on extracellular metabolites released by senescent fibroblasts supported a shift toward glycolysis compared to young cells 19 although this shift was not consistently observed in different cell types as observed by Delfarah et al, who found unaltered glucose consumption and lactate secretion in senescent human mammary epithelial cells 37 . Conversely, numerous studies reported that a high glycolysis rate, accompanied by elevated OXPHOS activity, was specific to oncogene-induced senescence 22 34 38 .…”

Section: Discussionmentioning

confidence: 92%

Delineating the heterogeneity of senescence-induced-functional alterations in hepatocytes

Kumar,

Hassan,

Tacke

et al. 2023

Preprint

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Background and Aim: Cellular senescence of hepatocytes involves permanent cell cycle arrest, disrupted cellular bioenergetics, resistance to cell death, and the release of pro-inflammatory cytokines. This zombie-like state perpetuates harmful effects on tissues and holds potential implications for liver disease progression. Remarkably, senescence exhibits heterogeneity, stemming from two crucial factors: the inducing stressor and the cell type. As such, our present study endeavors to characterize stressor-specific changes in senescence phenotype, its related molecular patterns, and cellular bioenergetics in primary mouse hepatocytes (PMH) and hepatocyte-derived liver organoids (HepOrgs). Methods: PMH, isolated by collagenase-perfused mouse liver (C57B6/J; 18-23 weeks), were cultured overnight in Williams E-medium supplemented with 2% FBS, L-glutamine, and hepatocyte growth supplements. HepOrgs were developed by culturing cells in a 3D matrix for two weeks. The senescence was induced by DNA damage (doxorubicin, cisplatin, and etoposide), oxidative stress (H2O2, and ethanol), and telomere inhibition (BIBR-1532), p53 activation (nutlin-3a), DNA methyl transferase inhibition (5-azacitidine), and metabolism inhibitors (galactosamine and hydroxyurea). SA-β galactosidase activity, immunofluorescence, immunoblotting, and senescence-associated secretory phenotype (SASP), and cellular bioenergetics were used to assess the senescence phenotype. Results: Each senescence inducer triggers a unique combination of senescence markers in hepatocytes. All senescence inducers, except hydroxyurea and ethanol, increased SA-beta galactosidase activity, the most commonly used marker for cellular senescence. Among the SASP factors, CCL2 and IL-10 were consistently upregulated, while Plasminogen activator inhibitor-1 exhibited global downregulation across all modes of senescence. Notably, DNA damage response was activated by DNA damage inducers. Cell cycle markers were most significantly reduced by doxorubicin, cisplatin, and galactosamine. Additionally, DNA damage-induced senescence shifted cellular bioenergetics capacity from glycolysis to oxidative phosphorylation. Conclusion: In our study, we demonstrated that each senescence inducer activates a unique combination of senescence markers in PMH. Doxorubicin demonstrated the highest efficacy in inducing senescence, followed by cisplatin and H2O2, with no impact on apoptosis. Each inducer prompted DNA damage response and mitochondrial dysfunction, independent of MAPK/AKT.

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“…1A). We then used liquid chromatography-mass spectrometry (LC-MS) metabolomics to measure changes in metabolite abundance and TCA cycle flux upon glucose deprivation [25][26][27] . Using two sensitive cell lines (LN18, T98) and one medium resistant cell line (LN229), we quantified levels of 97 polar metabolites in cells cultured with or without glucose for 3 h (Supporting Table 1).…”

Section: Metabolomics Identifies Accumulation Of L-cystine and L-cystmentioning

confidence: 99%

NADPH consumption by L-cystine reduction creates a metabolic vulnerability upon glucose deprivation

Joly¹,

Delfarah²,

Phung³

et al. 2019

Preprint

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The consequences of metabolic reprogramming in cancer can include an increased dependence on metabolic substrates such as glucose for survival. As such, the vulnerability of cancer cells to glucose deprivation creates an attractive opportunity for therapeutic intervention. Because it is not possible to starve tumors of glucose in vivo, we sought to identify the mechanisms regulating cancer cell death upon glucose deprivation and then design combinations of inhibitors to mimic glucose deprivation-induced cell death. Using metabolomic profiling, we found that cells undergoing glucose deprivationinduced cell death exhibited dramatic accumulation of intracellular L-cysteine and its oxidized dimer, L-cystine, and depletion of the antioxidant glutathione. Building on this observation, we show that glucose deprivation-induced cell death is driven not by lack of glucose but rather by L-cystine import. Following glucose deprivation, the import of Lcystine and subsequent reduction to L-cysteine depleted both NADPH and glutathione, thereby allowing toxic accumulation of reactive oxygen species. Consistent with this model, we found that the glutamate/cystine antiporter, xCT, was required for sensitivity to glucose deprivation. We searched for glycolytic enzymes whose expression is essential for survival of cancer cells with high xCT expression and identified the glucose transporter GLUT1. We therefore tested a drug combination co-targeting GLUT1 and glutathione synthesis and found that these drugs induced synthetic lethal cell death in high xCTexpressing cell lines susceptible to glucose deprivation. These results indicate that cotargeting GLUT1 and glutathione synthesis is a potential therapeutic approach in tumors dependent on glucose for survival. L-cystine drives glucose deprivation-induced cell deathIntroduction:

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Inhibition of nucleotide synthesis promotes replicative senescence of human mammary epithelial cells

Cited by 4 publications

References 63 publications

Tissue-Specific Landscape of Metabolic Dysregulation during Ageing

Tissue-Specific Landscape of Metabolic Dysregulation during Ageing

Delineating the heterogeneity of senescence-induced-functional alterations in hepatocytes

NADPH consumption by L-cystine reduction creates a metabolic vulnerability upon glucose deprivation

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