Gout and pseudo-gout-related crystals promote GLUT1-mediated glycolysis that governs NLRP3 and interleukin-1β activation on macrophages

ObjectiveMacrophage activation by monosodium urate (MSU) and calcium pyrophosphate (CPP) crystals mediates an interleukin (IL)-1β-dependent inflammation during gout and pseudo-gout flare, respectively. Since metabolic reprogramming of macrophages goes along with inflammatory responses dependently on stimuli and tissue environment, we aimed to decipher the role of glycolysis and oxidative phosphorylation in the IL-1β-induced microcrystal response.MethodsBriefly, an in vitro study (metabolomics and real-time extracellular flux analysis) on MSU and CPP crystal-stimulated macrophages was performed to demonstrate the metabolic phenotype of macrophages. Then, the role of aerobic glycolysis in IL-1β production was evaluated, as well in vitro as in vivo using 18F-fluorodeoxyglucose positron emission tomography imaging and glucose uptake assay, and molecular approach of glucose transporter 1 (GLUT1) inhibition.ResultsWe observed that MSU and CPP crystals led to a metabolic rewiring toward the aerobic glycolysis pathway explained by an increase in GLUT1 plasma membrane expression and glucose uptake on macrophages. Also, neutrophils isolated from human synovial fluid during gout flare expressed GLUT1 at their plasma membrane more frequently than neutrophils isolated from bloodstream. Both glucose deprivation and treatment with either 2-deoxyglucose or GLUT1 inhibitor suppressed crystal-induced NLRP3 activation and IL-1β production, and microcrystal inflammation in vivo.ConclusionIn conclusion, we demonstrated that GLUT1-mediated glucose uptake is instrumental during the inflammatory IL-1β response induced by MSU and CPP crystals. These findings open new therapeutic paths to modulate crystal-related inflammation.

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AL355338 acts as an oncogenic lncRNA by interacting with protein ENO1 to regulate EGFR/AKT pathway in NSCLC

Cancer Cell International ◽

10.1186/s12935-021-02232-z ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Qian Hua ◽

Dongliang Wang ◽

Lin Zhao ◽

Zhihui Hong ◽

Kairu Ni ◽

...

Keyword(s):

Aerobic Glycolysis ◽

Transcript Abundance ◽

Metabolic Reprogramming ◽

Clinical Samples ◽

Abnormal Metabolism ◽

Considerable Morbidity

Abstract Background Non-small cell lung cancer (NSCLC) is a malignancy with considerable morbidity and mortality. Abnormal metabolism is a hallmark of cancer; however, the mechanism of glycolysis regulation in NSCLC progression is not completely understood. Recent studies suggest that some dysregulated long non-coding RNAs (lncRNAs) play important roles in tumor metabolic reprogramming. Methods To identify glycolysis-associated-lncRNAs in NSCLC, we compared RNA-sequencing results between high 18F-fluorodeoxyglucose (FDG)-uptake NSCLC tissues and paired paratumor tissues. The transcript abundance of AL355338 in 80 pairs of clinical samples was evaluated by quantitative real-time PCR assay and fluorescence in situ hybridization. The biological role of AL355338 on NSCLC cells were evaluated by functional experiments in vitro and in vivo. Moreover, RNA pull-down, mass spectrometry and RNA immunoprecipitation (RIP) assays were used to identify the protein interacted with AL355338. Co-immunoprecipitation, in situ proximity ligation assays and western blotting were applied to define the potential downstream pathways of AL355338. Results AL355338 was an upregulated glycolysis-associated lncRNA in NSCLC. Functional assays revealed that AL355338 was critical for promoting aerobic glycolysis and NSCLC progression. Mechanistic investigations showed that AL355338 directly bound with alpha-enolase (ENO1) and enhanced the protein’s stability by modulating its degradation and ubiquitination. A positive correlation was observed between AL355338 and ENO1 in NSCLC, and ENO1 was subsequently confirmed to be responsible for the oncogenic role of AL355338. Furthermore, AL355338 was capable of modulating ENO1/EGFR complex interaction and further activating EGFR-AKT signaling. Conclusions This study indicates that AL355338 confers an aggressive phenotype to NSCLC, and targeting it might be an effective therapeutic strategy.

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TGF-βRII regulates glucose metabolism in oral cancer-associated fibroblasts via promoting PKM2 nuclear translocation

Cell Death Discovery ◽

10.1038/s41420-021-00804-6 ◽

2022 ◽

Vol 8 (1) ◽

Author(s):

Fanglong Wu ◽

Shimeng Wang ◽

Qingxiang Zeng ◽

Junjiang Liu ◽

Jin Yang ◽

...

Keyword(s):

Glucose Metabolism ◽

Oral Cancer ◽

Nuclear Translocation ◽

Aerobic Glycolysis ◽

Critical Role ◽

Metabolic Reprogramming ◽

Cancer Associated Fibroblasts

AbstractCancer-associated fibroblasts (CAFs) are highly heterogeneous and differentiated stromal cells that promote tumor progression via remodeling of extracellular matrix, maintenance of stemness, angiogenesis, and modulation of tumor metabolism. Aerobic glycolysis is characterized by an increased uptake of glucose for conversion into lactate under sufficient oxygen conditions, and this metabolic process occurs at the site of energy exchange between CAFs and cancer cells. As a hallmark of cancer, metabolic reprogramming of CAFs is defined as reverse Warburg effect (RWE), characterized by increased lactate, glutamine, and pyruvate, etc. derived from aerobic glycolysis. Given that the TGF-β signal cascade plays a critical role in RWE mainly through metabolic reprogramming related proteins including pyruvate kinase muscle isozyme 2 (PKM2), however, the role of nuclear PKM2 in modifying glycolysis remains largely unknown. In this study, using a series of in vitro and in vivo experiments, we provide evidence that TGF-βRII overexpression suppresses glucose metabolism in CAFs by attenuating PKM2 nuclear translocation, thereby inhibiting oral cancer tumor growth. This study highlights a novel pathway that explains the role of TGF-βRII in CAFs glucose metabolism and suggests that targeting TGF-βRII in CAFs might represent a therapeutic approach for oral cancer.

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AL355338 acts as an oncogenic lncRNA by interacting with protein ENO1 to regulate EGFR/AKT pathway in NSCLC

10.21203/rs.3.rs-815448/v1 ◽

2021 ◽

Author(s):

Qian Hua ◽

Dongliang Wang ◽

Lin Zhao ◽

Zhihui Hong ◽

Kairu Ni ◽

...

Keyword(s):

Aerobic Glycolysis ◽

Transcript Abundance ◽

Metabolic Reprogramming ◽

Clinical Samples ◽

Abnormal Metabolism ◽

Considerable Morbidity

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Mitochondrial Genome Regulator TFAM Modulates Head and Neck Tumorigenesis Through Activation of Intracellular Metabolic Reprogramming and Oncogenic Effectors

10.21203/rs.3.rs-78709/v1 ◽

2020 ◽

Author(s):

Yi-Ta Hsieh ◽

Hsi-Feng Tu ◽

Muh-Hwa Yang ◽

Yi-Fen Chen ◽

Chien-Ling Huang ◽

...

Keyword(s):

Mitochondrial Genome ◽

Head And Neck ◽

Aerobic Glycolysis ◽

Survival Rates ◽

Cellular Level ◽

Metabolic Reprogramming ◽

Clinical Databases

Abstract Background: Mitochondrial defect is often observed in cancers while, in comparison with other metabolic cues, mitochondria mediated regulations in controlling tumorigenesis are less emphasized. Mitochondrial transcriptional factor A (TFAM) acts as a key regulatory factor to control mitochondrial DNA (mtDNA) replication and packing; the role of TFAM in modulating carcinogenesis, however, is controversial. Current study therefore aims to define TFAM mediated regulations in head and neck cancer (HNC) development. Methods: Multifaceted analyses in HNC cells genetically manipulated for TFAM were performed. Clinical correlations of TFAM and its downstream Electron Transport Chain (ETC) associated factors in regulating HNC progression were also examined in HNC specimens and different clinical databasesResults: At the cellular level, it was demonstrated that shRNA mediated TFAM silencing resulted in an enhanced cell proliferation, both in vitro and in vivo; in contrast, TFAM overexpression suppressed cell growth. Moreover, TFAM loss also facilitated cell migration and chemodrug resistance. At the molecular basis, TFAM mediated phenotypic changes could be resulting from metabolic reprogramming by directing HNC metabolism towards aerobic glycolysis, based on the detection of less respiratory capacity in accompany with greater extracellular acidification in response to TFAM loss. Interestingly, it was also found that TFAM loss upregulated ERK1/2 and Akt-mTORC-S6 signaling activity, revealing a potential "mitochondrion-to-cytoplasm" retrograde regulatory cue in controlling HNC malignancy. Clinical impact of TFAM and its downstream targets was further examined in clinical HNC tissues while the results showed that TFAM expression and mtDNA copy numbers were significant dropped in HNC tissues compared with their normal counterparts. By using clinical databases, HNC subjects with higher TFAM expression and less genetic alteration(s) exhibited better survival rates. Conclusion: Collectively, Current study uncovered a tumor suppressing role of TFAM and mitochondrial genome in determining HNC oncogenicity. This TFAM mediated regualtions are through intracellular metabolic reprogramming and mitochondria-to-cytoplasm cross-talk to activate oncogenic signals.

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Dysregulated GLUT1 May Be Involved in the Pathogenesis of Preeclampsia by Impairing Decidualization

10.21203/rs.3.rs-842160/v1 ◽

2021 ◽

Author(s):

Man Yang ◽

Hua Li ◽

Miaomiao Rong ◽

Hongya Zhang ◽

Linlin Hou ◽

...

Keyword(s):

Glucose Uptake ◽

Glucose Transporter ◽

Aerobic Glycolysis ◽

Lactate Production ◽

Target Prediction ◽

Mrna Levels ◽

Endometrial Stromal Cells ◽

Glucose Transporter 1

Abstract Background: Preeclampsia (PE), a hypertensive complication in pregnancy, is a major contributor to maternal and fetal morbidity and mortality. Thus far, the molecular mechanism underlying PE has not been investigated thoroughly. Glucose transporter 1 (GLUT1) is a central rate-limiting pump for glucose uptake and subsequent utilization. Our previous RNA-seq results demonstrated it was significantly downregulated in deciduas from severe PE patients. Therefore, we aimed to explore the role of GLUT1 in the occurrence of PE.Methods: In this study, GLUT1 levels were evaluated by quantitative PCR, Western blotting and immunohistochemical staining in severe preeclamptic deciduas. The levels of GLUT1 during decidualization were also studied in human endometrial stromal cells (hESCs). Moreover, the role of GLUT1 during decidualization was studied by GLUT1-siRNA treatment. Furthermore, we explored the regulatory role of miRNA in GLUT1 expression.Results: The expression of GLUT1 was significantly downregulated in the deciduas from severe PE patients. Additionally, the level of GLUT1 was substantially induced in hESCs during in vitro decidualization. Moreover, GLUT1 knockdown significantly reduced the mRNA levels of decidualization markers (IGFBP1 and PRL) and aerobic glycolysis-related genes (LDHA and MCT4), and decreased glucose uptake and lactate production. Furthermore, the levels of apoptotic genes P53, P21 and BAX increased whereas the levels of BCL2 decreased after GLUT1 knockdown. Target prediction results and luciferase analysis showed GLUT1 is one of the targets of miR-140-5p, which is partly responsible for the impaired GLUT1 level. Conclusion: These results demonstrate that GLUT1 exerts pivotal role in human decidualization by participating in glycolysis, and its deficiency may trigger aberrant glycolysis and thus leads to destructive decidualization, which may be a pathogenetic mechanism of PE. These data suggest GLUT1 might be a promising target for PE therapy.

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Cortistatin Is Not a Somatostatin Analogue but Stimulates Prolactin Release and Inhibits GH and ACTH in a Gender-Dependent Fashion: Potential Role of Ghrelin

Endocrinology ◽

10.1210/en.2011-1542 ◽

2011 ◽

Vol 152 (12) ◽

pp. 4800-4812 ◽

Cited By ~ 40

Author(s):

José Córdoba-Chacón ◽

Manuel D. Gahete ◽

Ana I. Pozo-Salas ◽

Antonio J. Martínez-Fuentes ◽

Luis de Lecea ◽

...

Keyword(s):

Metabolic Phenotype ◽

Somatostatin Analogue ◽

Growth Hormone Secretagogue Receptor ◽

Growth Hormone Secretagogue ◽

Ancestral Gene ◽

Ghrelin Receptor ◽

Physiological Relevance

Cortistatin (CST) and somatostatin (SST) evolve from a common ancestral gene and share remarkable structural, pharmacological, and functional homologies. Although CST has been considered as a natural SST-analogue acting through their shared receptors (SST receptors 1–5), emerging evidence indicates that these peptides might in fact exert unique roles via selective receptors [e.g. CST, not SST, binds ghrelin receptor growth hormone secretagogue receptor type 1a (GHS-R1a)]. To determine whether the role of endogenous CST is different from SST, we characterized the endocrine-metabolic phenotype of male/female CST null mice (cort−/−) at hypothalamic-pituitary-systemic (pancreas-stomach-adrenal-liver) levels. Also, CST effects on hormone expression/secretion were evaluated in primary pituitary cell cultures from male/female mice and female primates (baboons). Specifically, CST exerted an unexpected stimulatory role on prolactin (PRL) secretion, because both male/female cort−/− mice had reduced PRL levels, and CST treatment (in vivo and in vitro) increased PRL secretion, which could be blocked by a GHS-R1a antagonist in vitro and likely relates to the decreased success of female cort−/− in first-litter pup care at weaning. In contrast, CST inhibited GH and adrenocorticotropin-hormone axes in a gender-dependent fashion. In addition, a rise in acylated ghrelin levels was observed in female cort−/− mice, which were associated with an increase in stomach ghrelin/ghrelin O-acyl transferase expression. Finally, CST deficit uncovered a gender-dependent role of this peptide in the regulation of glucose-insulin homeostasis, because male, but not female, cort−/− mice developed insulin resistance. The fact that these actions are not mimicked by SST and are strongly gender dependent offers new grounds to investigate the hitherto underestimated physiological relevance of CST in the regulation of physiological/metabolic processes.

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Purinergic Modulation of Interleukin-1β Release from Microglial Cells Stimulated with Bacterial Endotoxin

Journal of Experimental Medicine ◽

10.1084/jem.185.3.579 ◽

1997 ◽

Vol 185 (3) ◽

pp. 579-582 ◽

Cited By ~ 342

Author(s):

Davide Ferrari ◽

Paola Chiozzi ◽

Simonetta Falzoni ◽

Stefania Hanau ◽

Francesco Di Virgilio

Keyword(s):

Plasma Membrane ◽

P2x7 Receptor ◽

Purinergic Receptor ◽

Present Report ◽

Physiological Role ◽

Bacterial Endotoxin ◽

Microglial Cells

Microglial cells express a peculiar plasma membrane receptor for extracellular ATP, named P2Z/P2X7 purinergic receptor, that triggers massive transmembrane ion fluxes and a reversible permeabilization of the plasma membrane to hydrophylic molecules of up to 900 dalton molecule weight and eventual cell death (Di Virgilio, F. 1995. Immunol. Today. 16:524–528). The physiological role of this newly cloned (Surprenant, A., F. Rassendren, E. Kawashima, R.A. North and G. Buell. 1996. Science (Wash. DC). 272:735–737) cytolytic receptor is unknown. In vitro and in vivo activation of the macrophage and microglial cell P2Z/P2X7 receptor by exogenous ATP causes a large and rapid release of mature IL-1β. In the present report we investigated the role of microglial P2Z/P2X7 receptor in IL-1β release triggered by LPS. Our data suggest that LPS-dependent IL-1β release involves activation of this purinergic receptor as it is inhibited by the selective P2Z/P2X7 blocker oxidized ATP and modulated by ATP-hydrolyzing enzymes such as apyrase or hexokinase. Furthermore, microglial cells release ATP when stimulated with LPS. LPS-dependent release of ATP is also observed in monocyte-derived human macrophages. It is suggested that bacterial endotoxin activates an autocrine/paracrine loop that drives ATP-dependent IL-1β secretion.

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Let-7a regulates EV secretion and mitochondrial oxidative phosphorylation by targeting SNAP23 in colorectal cancer

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-020-01813-6 ◽

2021 ◽

Vol 40 (1) ◽

Author(s):

You Dong Liu ◽

Xiao Peng Zhuang ◽

Dong Lan Cai ◽

Can Cao ◽

Qi Sheng Gu ◽

...

Keyword(s):

Oxidative Phosphorylation ◽

Metabolic Reprogramming ◽

Luciferase Reporter ◽

Mitochondrial Oxidative Phosphorylation ◽

In Vivo Assays ◽

Reporter Assays ◽

Extracellular Mirna

Abstract Background MicroRNAs (miRNAs) are abundant in tumor-derived extracellular vesicles (EVs) and the functions of extracellular miRNA to recipient cells have been extensively studied with tumorigenesis. However, the role of miRNA in EV secretion from cancer cells remains unknown. Methods qPCR and bioinformatics analysis were applied for determining extracellular let-7a expression from CRC patient serum and cells. Nanosight particle tracking analysis was performed for investigating the effect of let-7a on EV secretion. Luciferase reporter assays was used for identifying targeted genes synaptosome-associated protein 23 (SNAP23). In vitro and in vivo assays were used for exploring the function of let-7a/SNAP23 axis in CRC progression. Bioenergetic assays were performed for investigating the role of let-7a/SNAP23 in cellular metabolic reprogramming. Results let-7a miRNA was elevated in serum EVs from CRC patients and was enriched in CRC cell-derived EVs. We determined that let-7a could suppress EV secretion directly targeting SNAP23. In turn, SNAP23 promotes EV secretion of let-7a to downregulate the intracellular let-7a expression. In addition, we found a novel mechanism of let-7a/SNAP23 axis by regulating mitochondrial oxidative phosphorylation (OXPHOS) through Lin28a/SDHA signaling pathway. Conclusions Let-7a plays an essential role in not only inhibiting EV secretion, but also suppressing OXPHOS through SNAP23, resulting in the suppression of CRC progression, suggesting that let-7a/SNAP23 axis could provide not only effective tumor biomarkers but also novel targets for tumor therapeutic strategies.

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TOP1MT deficiency promotes GC invasion and migration via the enhancements of LDHA expression and aerobic glycolysis

Endocrine Related Cancer ◽

10.1530/erc-17-0058 ◽

2017 ◽

Vol 24 (11) ◽

pp. 565-578 ◽

Cited By ~ 7

Author(s):

Hongqiang Wang ◽

Rui Zhou ◽

Li Sun ◽

Jianling Xia ◽

Xuchun Yang ◽

...

Keyword(s):

Cancer Progression ◽

Topoisomerase I ◽

Target Genes ◽

Aerobic Glycolysis ◽

Lactate Production ◽

Migration And Invasion ◽

Glucose Transporter 1 ◽

Mesenchymal Transition

Aerobic glycolysis plays an important role in cancer progression. New target genes regulating cancer aerobic glycolysis must be explored to improve patient prognosis. Mitochondrial topoisomerase I (TOP1MT) deficiency suppresses glucose oxidative metabolism but enhances glycolysis in normal cells. Here, we examined the role of TOP1MT in gastric cancer (GC) and attempted to determine the underlying mechanism. Using in vitro and in vivo experiments and analyzing the clinicopathological characteristics of patients with GC, we found that TOP1MT expression was lower in GC samples than in adjacent nonmalignant tissues. TOP1MT knockdown significantly promoted GC migration and invasion in vitro and in vivo. Importantly, TOP1MT silencing increased glucose consumption, lactate production, glucose transporter 1 expression and the epithelial-mesenchymal transition (EMT) in GC. Additionally, regulation of glucose metabolism induced by TOP1MT was significantly associated with lactate dehydrogenase A (LDHA) expression. A retrospective analysis of clinical data from 295 patients with GC demonstrated that low TOP1MT expression was associated with lymph node metastasis, recurrence and high mortality rates. TOP1MT deficiency enhanced glucose aerobic glycolysis by stimulating LDHA to promote GC progression.

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Multinucleated rat alveolar macrophages express functional receptors for calcitonin

AJP Renal Physiology ◽

10.1152/ajprenal.1991.261.6.f1026 ◽

1991 ◽

Vol 261 (6) ◽

pp. F1026-F1032 ◽

Cited By ~ 1

Author(s):

A. Vignery ◽

M. J. Raymond ◽

H. Y. Qian ◽

F. Wang ◽

S. A. Rosenzweig

Keyword(s):

Plasma Membrane ◽

Alveolar Macrophages ◽

Giant Cells ◽

Mononuclear Phagocytes ◽

Inflammatory Reactions ◽

Calcitonin Gene ◽

Novel Model

The fusion of mononuclear phagocytes occurs spontaneously in vivo and leads to the differentiation of either multinucleated giant cells or osteoclasts in chronic inflammatory sites or in bone, respectively. Although osteoclasts are responsible for resorbing bone, the functional role of giant cells in chronic inflammatory reactions and tumors remains poorly understood. We recently reported that the plasma membrane of multinucleated macrophages is, like that of osteoclasts, enriched in Na-K-adenosinetriphosphatases (ATPases). We also observed that the localization of their Na-K-ATPases is restricted to the nonadherent domain of the plasma membrane of cells both in vivo and in vitro, thus imposing a functional polarity on their organization. By following this observation, we wished to investigate whether these cells also expressed, like osteoclasts, functional receptors for calcitonin (CT). To this end, alveolar macrophages were fused in vitro, and both their structural and functional association with CT was analyzed and compared with those of mononucleated peritoneal and alveolar macrophages. Evidence is presented that multinucleated alveolar macrophages express a high copy number of functional receptors for CT. Our results also indicate that alveolar macrophages, much like peritoneal, express functional receptors for calcitonin gene-related peptide. It is suggested that multinucleated rat alveolar macrophages offer a novel model system to study CT receptors and that calcitonin may control local immune reactions where giant cells differentiate.

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