title: Glutamine fuels a vicious cycle of autophagy in the tumor stroma and oxidative mitochondrial metabolism in epithelial cancer cells: implications for preventing chemotherapy resistance.
abstract: Glutamine metabolism is crucial for cancer cell growth via the generation of intermediate molecules in the tricarboxylic acid (TCA) cycle, antioxidants and ammonia. The goal of the current study was to evaluate the effects of glutamine on metabolism in the breast cancer tumor microenvironment, with a focus on autophagy and cell death in both epithelial and stromal compartments. For this purpose, MCF7 breast cancer cells were cultured alone or co-cultured with non-transformed fibroblasts in media containing high glutamine and low glucose (glutamine +) or under control conditions, with no glutamine and high glucose (glutamine -). Here, we show that MCF7 cells maintained in co-culture with glutamine display increased mitochondrial mass, as compared with control conditions. Importantly, treatment with the autophagy inhibitor chloroquine abolishes the glutamine-induced augmentation of mitochondrial mass. It is known that loss of caveolin-1 (Cav-1) expression in fibroblasts is associated with increased autophagy and an aggressive tumor microenvironment. Here, we show that Cav-1 downregulation which occurs in fibroblasts maintained in co-culture specifically requires glutamine. Interestingly, glutamine increases the expression of autophagy markers in fibroblasts, but decreases expression of autophagy markers in MCF7 cells, indicating that glutamine regulates the autophagy program in a compartment-specific manner. Functionally, glutamine protects MCF7 cells against apoptosis, via the upregulation of the anti-apoptotic and anti-autophagic protein TIGAR. Also, we show that glutamine cooperates with stromal fibroblasts to confer tamoxifen-resistance in MCF7 cancer cells. Finally, we provide evidence that co-culture with fibroblasts (1) promotes glutamine catabolism, and (2) decreases glutamine synthesis in MCF7 cancer cells. Taken together, our findings suggest that autophagic fibroblasts may serve as a key source of energy-rich glutamine to fuel cancer cell mitochondrial activity, driving a vicious cycle of catabolism in the tumor stroma and anabolic tumor cell expansion.
authors.affiliation: The Jefferson Stem Cell Biology and Regenerative Medicine Center, Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA.
authors.forename: Ying-Hui
authors.initials: YH
authors.lastname: Ko
authors.forename: Zhao
authors.initials: Z
authors.lastname: Lin
authors.forename: Neal
authors.initials: N
authors.lastname: Flomenberg
authors.forename: Richard G
authors.initials: RG
authors.lastname: Pestell
authors.forename: Anthony
authors.initials: A
authors.lastname: Howell
authors.forename: Federica
authors.initials: F
authors.lastname: Sotgia
authors.forename: Michael P
authors.initials: MP
authors.lastname: Lisanti
authors.forename: Ubaldo E
authors.initials: UE
authors.lastname: Martinez-Outschoorn
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D051017:Apoptosis Regulatory Proteins; D001343:Autophagy / Q000187:drug effects* / Q000502:physiology; D051242:Caveolin 1 / Q000096:biosynthesis / Q000172:deficiency / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D002738:Chloroquine / Q000494:pharmacology; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D005347:Fibroblasts / Q000378:metabolism; D005973:Glutamine / Q000096:biosynthesis / Q000378:metabolism / Q000494:pharmacology*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000096:biosynthesis; D061986:MCF-7 Cells; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D009369:Neoplasms / Q000378:metabolism* / Q000473:pathology; D010085:Oxidative Phosphorylation / Q000187:drug effects; D010744:Phosphoric Monoester Hydrolases; D017154:Stromal Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D013629:Tamoxifen / Q000494:pharmacology; D059016:Tumor Microenvironment
chemical_list: D051017:Apoptosis Regulatory Proteins; D051242:Caveolin 1; D047908:Intracellular Signaling Peptides and Proteins; D013629:Tamoxifen; D005973:Glutamine; D002738:Chloroquine; D010744:Phosphoric Monoester Hydrolases; C512708:TIGAR protein, human