Ontology highlight
ABSTRACT:
PROVIDER: PRJNA1122213 | ENA |
REPOSITORIES: ENA
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Project description:The non-essential amino acid serine is a critical nutrient for cancer cells due to its diverse biosynthetic functions. While some tumors can synthesize serine de novo, others are auxotrophic and therefore reliant on serine uptake. Importantly, despite several transporters being known to be capable of transporting serine, the transporter(s) that mediate serine uptake in cancer cells are not known. Here, we characterize the amino acid transporter ASCT2 (SLC1A5) as a major contributor to serine uptake in cancer cells. ASCT2 is well-known as a glutamine transporter in cancer, and our work demonstrates that serine and glutamine compete for uptake through ASCT2. We further show that ASCT2-mediated serine uptake is essential for purine nucleotide biosynthesis and that ERα promotes serine uptake by directly activating SLC1A5 transcription. Together, our work defines an additional important role for ASCT2 as a serine transporter in cancer and evaluates ASCT2 as a potential therapeutic target.
2024-06-11 | GSE269606 | GEO
Project description:ASCT2 is a major contributor to serine uptake in cancer cells
| PRJNA1122640 | ENA
Project description:Triple-negative breast cancer (TNBC) relies on glutamine uptake by the transporter ASCT2 to sustain their unique glutamine metabolism and growth. Despite previous data showing cell growth inhibition after ASCT2 knockdown, ASCT2 CRISPR knockout was well-tolerated by breast cancer cell lines. Despite the loss of a glutamine transporter and low rate of glutamine uptake, intracellular glutamine steady state levels were higher in ASCT2 knockout compared to control TNBC cells. Proteomics data revealed upregulation of macropinocytosis, reduction in glutamine efflux and glutamine synthesis in ASCT2 knockout cells. Loss of ASCT2 in TNBC cell line HCC1806 induced a 5-10-fold increase in macropinocytosis across 5 separate ASCT2 knockout clones, compared to a modest 2-fold increase in the shRNA ASCT2 knockdown. By comparison, ASCT2 knockout impaired cell proliferation in a non-macropinocytic breast cancer cell line, HCC1569. These data suggest that macropinocytosis provides a novel resistance mechanism to strategies targeting glutamine uptake alone. Despite this adaptation, TNBC cells continue to rely on glutamine metabolism for their growth, which suggests therapeutic targeting may need to focus on downstream glutamine metabolism pathways.
2024-09-15 | GSE256218 | GEO
Project description:Although a nonessential amino acid in normal cells, the demand for glutamine is dramatically increased throughout malignant transformation, supporting a range of metabolic processes including mitochondrial ATP production, protein synthesis, purine and pyrimidine biosynthesis. We previously showed that triple-negative breast cancer (TNBC) cells rely on glutamine uptake by the amino acid transporter ASCT2 to sustain their unique glutamine metabolism, thereby supporting in vitro growth and in vivo tumour formation. However, it is known that TNBC cells can also utilise non-transporter mediated nutrient uptake facilitated by processes such as macropinocytosis. We examined proliferation and colony forming ability of human breast cancer cell lines after ASCT2 CRISPR/Cas9 knockout (clonal and polyclonal populations) and shRNA knockdown. Proteomics and mRNAseq analysis further examined cellular and adaptive changes to ASCT2 knockout. Cellular changes were further analysed by western blotting, with macropinocytosis examined using 70kDa dextran-FITC uptake. Metabolic changes were assessed using targeted metabolomics approaches including 13C-labelled substrate tracing and liquid chromatography coupled tandem-mass spectrometry (LC-MS/MS) to determine intracellular levels of key tricarboxylic acid (TCA) cycle intermediates, glycolytic metabolites, fatty acid precursors, nucleotides, and amino acids in human TNBC cell lines in vitro. Despite our previous data showing a significant reduction in cell growth after ASCT2 knockdown, ASCT2 knockout was well-tolerated by both TNBC and Luminal A breast cancer cell lines, with proliferation rates similar to non-targeted CRISPR/Cas9 control cells. This adaptation to knockout was not due to the high glutamine levels present in culture media, as the knockout cells could be cloned in media containing physiological 0.5 mM glutamine. Previous data have shown that TNBC cell lines can undergo constitutive macropinocytosis, and that this could be enhanced when cells are cultured in low nutrient conditions. Indeed, not only did the TNBC cell line HCC1806 undergo constitutive macropinocytosis, the amount of macropinocytosis was significantly enhanced (5-10 fold) in 5 separate ASCT2 knockout clones. By comparison, the ASCT2 knockdown cell line, which have a significant proliferation deficit, showed a modest 2-fold increase in macropinocytosis. Despite in-depth analysis of gene and protein levels by mRNAseq and proteomics, ASCT2 knockout cells did not display a significant alteration in macropinocytic gene expression, but instead showed a substantial upregulation of Ser473-Akt phosphorylation which may drive the adaptive macropinocytosis in TNBC. These data suggest that the constitutive macropinocytosis present in TNBC cell lines provides a novel resistance mechanism to strategies targeting glutamine uptake alone. Despite this adaptation, TNBC cells continue to rely on glutamine, however therapeutic targeting may need to focus on other unique TNBC metabolic pathways such as single-pass glutaminolysis, which couples glutamine and glucose metabolism together.
2024-09-16 | PXD053894 | Pride
Project description:To determine the global effects of ASCT2 inhibition, we used next generation sequencing to determine mRNA expression changes in PC-3 cells treated with BenSer or GPNA for 48 h. Examination of two different ASCT2 inhibitors BenSer and GPNA in prostate cancer cell line PC-3.
2015-04-10 | E-GEOD-65112 | biostudies-arrayexpress
Project description:Glutamine-dependence of cancer cells reduces local glutamine availability, which hinders anti-tumor T-cell functionality and facilitates immune evasion. We thus speculated that glutamine deprivation might be limiting efficacy of CAR T-cell therapies in cancer patients. We have seen that antigen-specific T cells are unable to proliferate or produce IFN-γ in response to antigen stimulation when glutamine concentration is limited. Using multiple myeloma (MM) as a glutamine-dependent disease model, we found that murine CAR-T cells selectively targeting BCMA in MM cells were sensitive to glutamine deprivation. However, CAR-T cells engineered to increase glutamine uptake by expression of the glutamine transporter Asct2 exhibited enhanced proliferation and responsiveness to antigen stimulation, increased production of IFN-, and heightened cytotoxic activity, even under conditions of low glutamine concentration. Mechanistically, Asct2 overexpression reprogrammed CAR-T cell metabolic fitness, improving basal oxygen consumption rate and glycolytic function that enhanced CAR-T cell persistence in vivo. Accordingly, expression of Asct2 increased the efficacy of BCMA CAR-T cells in syngeneic and genetically-engineered mouse models of MM, which prolonged mouse survival. In patients, reduced expression of Asct2 by MM cells predicted poor outcome to combined immunotherapy and BCMA-CAR T-cell therapy. Our results indicate that reprogramming glutamine metabolism may enhance anti-tumor CAR T-cell functionality in multiple myeloma. This approach may also be effective for other cancers that depend on glutamine as a key energy source and metabolic hallmark.
2025-08-21 | GSE299766 | GEO
Project description:Head and neck cancer with its major subsite, oral squamous cell carcinoma (OSCC), is a devastating malignancy with limited treatment options. Given that recent advances in nutritional interventions have been recognized as potential strategies for targeting cancer, we investigated the role of dietary serine restriction in OSCC cell fate decisions. We now report that human OSCC cells behave like serine auxotrophs, depending on exogenous serine to promote growth and sustain plastic cell states, such as cancer stem cells and cells with EMT features. We show that restriction of serine induces endogenous serine synthesis and generates alpha-ketoglutarate (aKG), a co-substrate for JMJD3 nuclear dioxygenase, which demethylates H3K27me3 and de-represses differentiation genes. This is balanced by the loss of H3K4me3 and inhibition of stemness genes. Likewise, dietary serine restriction inhibits orthotopic OSCC growth in syngeneic mice. Our work provides the basis for including precision nutrition to advance treatments for OSCC patients.
2026-09-02 | GSE286465 | GEO
Project description:Cancer cells rely on metabolic reprogramming to sustain the prodigious energetic requirements for rapid growth and proliferation. Glutamine metabolism is frequently dysregulated in cancers and is being exploited as a potential therapeutic target. In current study, we identified TARBP1 (TAR (HIV-1) RNA Binding Protein 1) as a novel driver gene critical for glutamine metabolic reprogramming in tumor through the CRISPRi/Cas9 screening. Our in vivo and in vitro assays demonstrated that TARBP1 is the methyltransferase of Guanosine 2'-O-methylation targeting position 18 (G18) of tRNAGln (TTG/CTG) and tRNASer (TGA/GCT), and loss of Gm18 modification diminishes the stability of tRNAs. Therefore, TARBP1 is critical for maintaining efficient translation of mRNA, in particular the glutamine transportor-ASCT2 (also known as SCL1A5). Importantly, TARBP1 is frequently amplified and overexpressed in HCC, consequentially promotes the protein synthesis of ASCT2 and glutamine import to fuel the growth of cancer cell, which is associated with poor patient survival. Taken together, this study reveals the critical role of TARBP1 in HCC progression through glutamine metabolic reprogramming and provides a potential target for tumor therapy.
2024-06-23 | GSE236368 | GEO
Project description:Adipocyte serine uptake curbs ROS generation and visceral adiposity
| PRJNA557596 | ENA
Project description:Head and neck cancer with its major subsite, oral squamous cell carcinoma (OSCC), is a devastating malignancy with limited treatment options. Given that recent advances in nutritional interventions have been recognized as potential strategies for targeting cancer, we investigated the role of dietary serine restriction in OSCC cell fate decisions. We now report that human OSCC cells behave like serine auxotrophs, depending on exogenous serine to promote growth and sustain plastic cell states, such as cancer stem cells and cells with EMT features. We show that restriction of serine induces endogenous serine synthesis and generates alpha-ketoglutarate (aKG), a co-substrate for JMJD3 nuclear dioxygenase, which demethylates H3K27me3 and de-represses differentiation genes. This is balanced by the loss of H3K4me3 and inhibition of stemness genes. Likewise, dietary serine restriction inhibits orthotopic OSCC growth in syngeneic mice. Our work provides the basis for including precision nutrition to advance treatments for OSCC patients.
2026-09-02 | GSE286464 | GEO