The p53 Pathway and Metabolism: The Tree That Hides the Forest

The p53 pathway is functionally inactivated in most, if not all, human cancers. The p53 protein is a central effector of numerous stress-related molecular cascades. p53 controls a safeguard mechanism that prevents accumulation of abnormal cells and their transformation by regulating DNA repair, cell cycle progression, cell death, or senescence. The multiple cellular processes regulated by p53 were more recently extended to the control of metabolism and many studies support the notion that perturbations of p53-associated metabolic activities are linked to cancer development, as well as to other pathophysiological conditions including aging, type II diabetes, and liver disease. Although much less documented than p53 metabolic activities, converging lines of evidence indicate that other key components of this tumor suppressor pathway are also involved in cellular metabolism through p53-dependent as well as p53-independent mechanisms. Thus, at least from a metabolic standpoint, the p53 pathway must be considered as a non-linear pathway, but the complex metabolic network controlled by these p53 regulators and the mechanisms by which their activities are coordinated with p53 metabolic functions remain poorly understood. In this review, we highlight some of the metabolic pathways controlled by several central components of the p53 pathway and their role in tissue homeostasis, metabolic diseases, and cancer

Simultaneous Measurement of Metabolite Concentration and Isotope Incorporation by Mass Spectrometry

International audienceStudies of the topology, functioning, and regulation of metabolic systems are based on two main types of information that can be measured by mass spectrometry the (absolute or relative) concentration of metabolites and their isotope incorporation in C-labeling experiments. These data are currently obtained from two independent experiments because the C-labeled internal standard (IS) used to determine the concentration of a given metabolite overlaps the C-mass fractions from which its C-isotopologue distribution (CID) is quantified. Here, we developed a generic method with a dedicated processing workflow to obtain these two sets of information simultaneously in a unique sample collected from a single cultivation, thereby reducing by a factor of 2 both the number of cultivations to perform and the number of samples to collect, prepare, and analyze. The proposed approach is based on an IS labeled with other isotope(s) that can be resolved from the C-mass fractions of interest. As proof-of-principle, we analyzed amino acids using a doubly labeled NC-cell extract as IS. Extensive evaluation of the proposed approach shows a similar accuracy and precision compared to state-of-the-art approaches. We demonstrate the value of this approach by investigating the dynamic response of amino acids metabolism in mammalian cells upon activation of the protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK), a key component of the unfolded protein response. Integration of metabolite concentrations and isotopic profiles reveals a reduced de novo biosynthesis of amino acids upon PERK activation. The proposed approach is generic and can be applied to other (micro)organisms, analytical platforms, isotopic tracers, or classes of metabolites

Calpain-6 controls the fate of sarcoma stem cells by promoting autophagy and preventing senescence

International audienceSarcomas are still unsolved therapeutic challenges. Cancer stem cells are believed to contribute to sarcoma development, but lack of specific markers prevents their characterization and targeting. Here, we show that calpain-6 expression is associated with cancer stem cell features. In mouse models of bone sarcoma, calpain-6-expressing cells have unique tumor-initiating and metastatic capacities. Calpain-6 levels are especially high in tumors that have been successfully propagated in mouse to establish patient-derived xenografts. We found that calpain-6 levels are increased by hypoxia in vitro and calpain-6 is detected within hypoxic areas in tumors. Furthermore, calpain-6 expression depends on the stem cell transcription network that involves Oct4, Nanog, and Sox2 and is activated by hypoxia. Calpain-6 knockdown blocks tumor development in mouse and induces depletion of the cancer stem cell population. Data from transcriptomic analyses reveal that calpain-6 expression in sarcomas inversely correlates with senescence markers. Calpain-6 knockdown suppresses hypoxia-dependent prevention of senescence entry and also promotion of autophagic flux. Together, our results demonstrate that calpain-6 identifies sarcoma cells with stem-like properties and is a mediator of hypoxia to prevent senescence, promote autophagy, and maintain the tumor-initiating cell population. These findings open what we believe is a novel therapeutic avenue for targeting sarcoma stem cells

Coinfections with SARS‐CoV‐2 variants and influenza virus during the 2019 Coronavirus disease pandemic in Burkina Faso: A surveillance study

Abstract Background and Aim Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) particularly the variants of concern coinfections with influenza is a public health concern in Africa. We aimed to characterize the SARS‐CoV‐2 variants and determine the rate of coinfections with influenza in Burkina Faso. Methods COVID‐19 surveillance study was conducted between August 2021 and January 2022 using reverse transcription polymerase chain reaction (RT‐PCR). Positive specimens were further screened for SARS‐CoV‐2 variants using the multiple variants real‐time PCR kits. In addition, influenza virus strains were detected by RT‐PCR in SARS‐CoV‐2 positive specimens using the CDC primers, probes, and protocols. Results Of 324 specimens assessed, the Omicron and Delta variants of SARS‐CoV‐2 were the most prevalent with 27.2% [95% confident interval (CI): 22.5–32.4] and 22.2% [95% CI: 17.9–27.2], respectively. The Beta and Gamma variants were detected in 4.3% [95% CI: 2.4–7.1] and 0.3% [95% CI: 0.0–1.7], respectively. Coinfections of Omicron and Beta variants were reported in 21.3% [95% CI: 17.0–26.2], Omicron and Delta variants in 1.2% [95% CI: 0.3–3.1] of specimens, and the Omicron–Gamma variants' coinfections in 0.6% [95% CI: 0.1–2.2]. One COVID‐19 specimen with an undetected SARS‐CoV‐2 variant was also tested positive for the seasonal influenza A (H3N2) virus. No cases of pandemic influenza A (H1N1)pdm09, seasonal A/H1N1, and influenza B were detected. Conclusions The current World Health Organization SARS‐CoV‐2 variants of concern were prevalent and their coinfections with influenza were uncommon. Continuous surveillance of both pathogens is, however, needed because of their public health implications

Targeting MDM2-dependent serine metabolism as a therapeutic strategy for liposarcoma

International audienceWell-differentiated and dedifferentiated liposarcomas (LPSs) are characterized by a systematic amplification of the MDM2 oncogene, which encodes a key negative regulator of the p53 pathway. The molecular mechanisms underlying MDM2 overexpression while sparing wild-type p53 in LPS remain poorly understood. Here, we show that the p53-independent metabolic functions of chromatin-bound MDM2 are exacerbated in LPS and mediate an addiction to serine metabolism that sustains nucleotide synthesis and tumor growth. Treatment of LPS cells with Nutlin-3A, a pharmacological inhibitor of the MDM2-p53 interaction, stabilized p53 but unexpectedly enhanced MDM2-mediated control of serine metabolism by increasing its recruitment to chromatin, likely explaining the poor clinical efficacy of this class of MDM2 inhibitors. In contrast, genetic or pharmacological inhibition of chromatin-bound MDM2 by SP141, a distinct MDM2 inhibitor triggering its degradation, or interfering with de novo serine synthesis, impaired LPS growth both in vitro and in clinically relevant patient-derived xenograft models. Our data indicate that targeting MDM2 functions in serine metabolism represents a potential therapeutic strategy for LPS

Mitochondrial metabolism supports resistance to IDH mutant inhibitors in acute myeloid leukemia

Mutations in IDH induce epigenetic and transcriptional reprogramming, differentiation bias, and susceptibility to mitochondrial inhibitors in cancer cells. Here, we first show that cell lines, PDXs, and patients with acute myeloid leukemia (AML) harboring an IDH mutation displayed an enhanced mitochondrial oxidative metabolism. Along with an increase in TCA cycle intermediates, this AML-specific metabolic behavior mechanistically occurred through the increase in electron transport chain complex I activity, mitochondrial respiration, and methylation-driven CEBPα-induced fatty acid β-oxidation of IDH1 mutant cells. While IDH1 mutant inhibitor reduced 2-HG oncometabolite and CEBPα methylation, it failed to reverse FAO and OxPHOS. These mitochondrial activities were maintained through the inhibition of Akt and enhanced activation of peroxisome proliferator-activated receptor-γ coactivator-1 PGC1α upon IDH1 mutant inhibitor. Accordingly, OxPHOS inhibitors improved anti-AML efficacy of IDH mutant inhibitors in vivo. This work provides a scientific rationale for combinatory mitochondrial-targeted therapies to treat IDH mutant AML patients, especially those unresponsive to or relapsing from IDH mutant inhibitors