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  1. Article ; Online: NADH-independent enzymatic assay to quantify extracellular and intracellular L-lactate levels.

    Bouchez, Cyrielle L / Daubon, Thomas / Mourier, Arnaud

    STAR protocols

    2022  Volume 3, Issue 2, Page(s) 101403

    Abstract: Lactate is a central metabolite in energy metabolism and is also involved in cell signaling and epigenetic regulations. Here, we describe an NADH-independent enzymatic assay allowing rapid, selective, and sensitive quantification of L-lactate down to the ...

    Abstract Lactate is a central metabolite in energy metabolism and is also involved in cell signaling and epigenetic regulations. Here, we describe an NADH-independent enzymatic assay allowing rapid, selective, and sensitive quantification of L-lactate down to the pmol range. We detail lactate extraction from intracellular and extracellular fractions, followed by total protein amount determination and enzymatic assay. This approach allows quantification of intracellular and extracellular L-lactate levels, validated by treating adherent and non-adherent cells with inhibitors of lactate transporters (MCT).
    MeSH term(s) Energy Metabolism ; Enzyme Assays ; Lactic Acid/metabolism ; Monocarboxylic Acid Transporters/metabolism ; NAD/metabolism
    Chemical Substances Monocarboxylic Acid Transporters ; NAD (0U46U6E8UK) ; Lactic Acid (33X04XA5AT)
    Language English
    Publishing date 2022-05-16
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 2666-1667
    ISSN (online) 2666-1667
    DOI 10.1016/j.xpro.2022.101403
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Mitochondrial Biogenesis and Mitochondrial Reactive Oxygen Species (ROS): A Complex Relationship Regulated by the cAMP/PKA Signaling Pathway.

    Bouchez, Cyrielle / Devin, Anne

    Cells

    2019  Volume 8, Issue 4

    Abstract: Mitochondrial biogenesis is a complex process. It requires the contribution of both the nuclear and the mitochondrial genomes and therefore cross talk between the nucleus and mitochondria. Cellular energy demand can vary by great length and it is now ... ...

    Abstract Mitochondrial biogenesis is a complex process. It requires the contribution of both the nuclear and the mitochondrial genomes and therefore cross talk between the nucleus and mitochondria. Cellular energy demand can vary by great length and it is now well known that one way to adjust adenosine triphosphate (ATP) synthesis to energy demand is through modulation of mitochondrial content in eukaryotes. The knowledge of actors and signals regulating mitochondrial biogenesis is thus of high importance. Here, we review the regulation of mitochondrial biogenesis both in yeast and in mammalian cells through mitochondrial reactive oxygen species.
    MeSH term(s) Animals ; Cyclic AMP/metabolism ; Cyclic AMP-Dependent Protein Kinases/metabolism ; Humans ; Mitochondria/metabolism ; Organelle Biogenesis ; Reactive Oxygen Species/metabolism ; Signal Transduction
    Chemical Substances Reactive Oxygen Species ; Cyclic AMP (E0399OZS9N) ; Cyclic AMP-Dependent Protein Kinases (EC 2.7.11.11)
    Language English
    Publishing date 2019-03-27
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2661518-6
    ISSN 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells8040287
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article: The Warburg Effect in Yeast: Repression of Mitochondrial Metabolism Is Not a Prerequisite to Promote Cell Proliferation.

    Bouchez, Cyrielle L / Hammad, Noureddine / Cuvellier, Sylvain / Ransac, Stéphane / Rigoulet, Michel / Devin, Anne

    Frontiers in oncology

    2020  Volume 10, Page(s) 1333

    Abstract: O. Warburg conducted one of the first studies on tumor energy metabolism. His early discoveries pointed out that cancer cells display a decreased respiration and an increased glycolysis proportional to the increase in their growth rate, suggesting that ... ...

    Abstract O. Warburg conducted one of the first studies on tumor energy metabolism. His early discoveries pointed out that cancer cells display a decreased respiration and an increased glycolysis proportional to the increase in their growth rate, suggesting that they mainly depend on fermentative metabolism for ATP generation. Warburg's results and hypothesis generated controversies that are persistent to this day. It is thus of great importance to understand the mechanisms by which cancer cells can reversibly regulate the two pathways of their energy metabolism as well as the functioning of this metabolism in cell proliferation. Here, we made use of yeast as a model to study the Warburg effect and its eventual function in allowing an increased ATP synthesis to support cell proliferation. The role of oxidative phosphorylation repression in this effect was investigated. We show that yeast is a good model to study the Warburg effect, where all parameters and their modulation in the presence of glucose can be reconstituted. Moreover, we show that in this model, mitochondria are not dysfunctional, but that there are fewer mitochondria respiratory chain units per cell. Identification of the molecular mechanisms involved in this process allowed us to dissociate the parameters involved in the Warburg effect and show that oxidative phosphorylation repression is not mandatory to promote cell growth. Last but not least, we were able to show that neither cellular ATP synthesis flux nor glucose consumption flux controls cellular growth rate.
    Language English
    Publishing date 2020-08-19
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2649216-7
    ISSN 2234-943X
    ISSN 2234-943X
    DOI 10.3389/fonc.2020.01333
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Refining the Role of Pyruvate Dehydrogenase Kinases in Glioblastoma Development.

    Larrieu, Claire M / Storevik, Simon / Guyon, Joris / Pagano Zottola, Antonio C / Bouchez, Cyrielle L / Derieppe, Marie-Alix / Tan, Tuan Zea / Miletic, Hrvoje / Lorens, James / Tronstad, Karl Johan / Daubon, Thomas / Røsland, Gro Vatne

    Cancers

    2022  Volume 14, Issue 15

    Abstract: Glioblastoma (GB) are the most frequent brain cancers. Aggressive growth and limited treatment options induce a median survival of 12-15 months. In addition to highly proliferative and invasive properties, GB cells show cancer-associated metabolic ... ...

    Abstract Glioblastoma (GB) are the most frequent brain cancers. Aggressive growth and limited treatment options induce a median survival of 12-15 months. In addition to highly proliferative and invasive properties, GB cells show cancer-associated metabolic characteristics such as increased aerobic glycolysis. Pyruvate dehydrogenase (PDH) is a key enzyme complex at the crossroads between lactic fermentation and oxidative pathways, finely regulated by PDH kinases (PDHKs). PDHKs are often overexpressed in cancer cells to facilitate high glycolytic flux. We hypothesized that targeting PDHKs, by disturbing cancer metabolic homeostasis, would alter GB progression and render cells vulnerable to additional cancer treatment. Using patient databases, distinct expression patterns of PDHK1 and PDHK2 in GB tissues were obvious. To disturb protumoral glycolysis, we modulated PDH activity through the genetic or pharmacological inhibition of PDHK in patient-derived stem-like spheroids. Striking effects of PDHKs inhibition using dichloroacetate were observed in vitro on cell morphology and metabolism, resulting in increased intracellular ROS levels and decreased proliferation and invasion. In vivo findings confirmed a reduction in tumor size and better survival of mice implanted with PDHK1 and PDHK2 knockout cells. Adding a radiotherapeutic protocol further resulted in a reduction in tumor size and improved mouse survival in our model.
    Language English
    Publishing date 2022-08-02
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2527080-1
    ISSN 2072-6694
    ISSN 2072-6694
    DOI 10.3390/cancers14153769
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: "Labile" heme critically regulates mitochondrial biogenesis through the transcriptional co-activator Hap4p in

    Bouchez, Cyrielle L / Yoboue, Edgar D / de la Rosa Vargas, Livier E / Salin, Bénédicte / Cuvellier, Sylvain / Rigoulet, Michel / Duvezin-Caubet, Stéphane / Devin, Anne

    The Journal of biological chemistry

    2020  Volume 295, Issue 15, Page(s) 5095–5109

    Abstract: Heme (iron protoporphyrin IX) is a well-known prosthetic group for enzymes involved in metabolic pathways such as oxygen transport and electron transfer through the mitochondrial respiratory chain. However, heme has also been shown to be an important ... ...

    Abstract Heme (iron protoporphyrin IX) is a well-known prosthetic group for enzymes involved in metabolic pathways such as oxygen transport and electron transfer through the mitochondrial respiratory chain. However, heme has also been shown to be an important regulatory molecule (as "labile" heme) for diverse processes such as translation, kinase activity, and transcription in mammals, yeast, and bacteria. Taking advantage of a yeast strain deficient for heme production that enabled controlled modulation and monitoring of labile heme levels, here we investigated the role of labile heme in the regulation of mitochondrial biogenesis. This process is regulated by the HAP complex in yeast. Using several biochemical assays along with EM and epifluorescence microscopy, to the best of our knowledge, we show for the first time that cellular labile heme is critical for the post-translational regulation of HAP complex activity, most likely through the stability of the transcriptional co-activator Hap4p. Consequently, we found that labile heme regulates mitochondrial biogenesis and cell growth. The findings of our work highlight a new mechanism in the regulation of mitochondrial biogenesis by cellular metabolites.
    MeSH term(s) CCAAT-Binding Factor/genetics ; CCAAT-Binding Factor/metabolism ; Hemin/metabolism ; Mitochondria/metabolism ; Organelle Biogenesis ; Oxygen Consumption ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae/metabolism ; Saccharomyces cerevisiae Proteins/genetics ; Saccharomyces cerevisiae Proteins/metabolism ; Signal Transduction
    Chemical Substances CCAAT-Binding Factor ; HAP4 protein, S cerevisiae ; Saccharomyces cerevisiae Proteins ; Hemin (743LRP9S7N)
    Language English
    Publishing date 2020-02-18
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1074/jbc.RA120.012739
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Uc I Zs.108: Show issues Location:
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  6. Article ; Online: Lactate dehydrogenases promote glioblastoma growth and invasion via a metabolic symbiosis.

    Guyon, Joris / Fernandez-Moncada, Ignacio / Larrieu, Claire M / Bouchez, Cyrielle L / Pagano Zottola, Antonio C / Galvis, Johanna / Chouleur, Tiffanie / Burban, Audrey / Joseph, Kevin / Ravi, Vidhya M / Espedal, Heidi / Røsland, Gro Vatne / Daher, Boutaina / Barre, Aurélien / Dartigues, Benjamin / Karkar, Slim / Rudewicz, Justine / Romero-Garmendia, Irati / Klink, Barbara /
    Grützmann, Konrad / Derieppe, Marie-Alix / Molinié, Thibaut / Obad, Nina / Léon, Céline / Seano, Giorgio / Miletic, Hrvoje / Heiland, Dieter Henrik / Marsicano, Giovanni / Nikolski, Macha / Bjerkvig, Rolf / Bikfalvi, Andreas / Daubon, Thomas

    EMBO molecular medicine

    2022  Volume 14, Issue 12, Page(s) e15343

    Abstract: Lactate is a central metabolite in brain physiology but also contributes to tumor development. Glioblastoma (GB) is the most common and malignant primary brain tumor in adults, recognized by angiogenic and invasive growth, in addition to its altered ... ...

    Abstract Lactate is a central metabolite in brain physiology but also contributes to tumor development. Glioblastoma (GB) is the most common and malignant primary brain tumor in adults, recognized by angiogenic and invasive growth, in addition to its altered metabolism. We show herein that lactate fuels GB anaplerosis by replenishing the tricarboxylic acid (TCA) cycle in absence of glucose. Lactate dehydrogenases (LDHA and LDHB), which we found spatially expressed in GB tissues, catalyze the interconversion of pyruvate and lactate. However, ablation of both LDH isoforms, but not only one, led to a reduction in tumor growth and an increase in mouse survival. Comparative transcriptomics and metabolomics revealed metabolic rewiring involving high oxidative phosphorylation (OXPHOS) in the LDHA/B KO group which sensitized tumors to cranial irradiation, thus improving mouse survival. When mice were treated with the antiepileptic drug stiripentol, which targets LDH activity, tumor growth decreased. Our findings unveil the complex metabolic network in which both LDHA and LDHB are integrated and show that the combined inhibition of LDHA and LDHB strongly sensitizes GB to therapy.
    MeSH term(s) Animals ; Mice ; Lactate Dehydrogenases ; Metabolomics ; Neoplasms ; Lactic Acid
    Chemical Substances Lactate Dehydrogenases (EC 1.1.-) ; Lactic Acid (33X04XA5AT)
    Language English
    Publishing date 2022-10-24
    Publishing country England
    Document type Journal Article
    ZDB-ID 2467145-9
    ISSN 1757-4684 ; 1757-4676
    ISSN (online) 1757-4684
    ISSN 1757-4676
    DOI 10.15252/emmm.202115343
    Database MEDical Literature Analysis and Retrieval System OnLINE

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