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Glioblastomas (GBM) account for almost half of the malignant central nervous system tumors and have the highest mortality rates with only about 5% of the adult patients surviving 5 years after diagnosis. Current therapy for GBM is... more
Glioblastomas (GBM) account for almost half of the malignant central nervous system tumors and have the highest mortality rates with only about 5% of the adult patients surviving 5 years after diagnosis. Current therapy for GBM is generally very aggressive, combining chemotherapy, radiotherapy, and surgical removal and increased the life expectancy only from 11 to 14 months, but, almost no significant advances were made in the last decades. Taking this into account, more effective compounds and new strategies, such as chronotherapeutic ones, need to be developed. The novel drug 1A-116 specifically blocks the interaction of the Rho GTPase Rac1 with some of its activators (GEFs proteins, such as Tiam1) inhibiting cell motility and proliferation of GBM. To optimize the effectiveness of 1A-116 a chronotherapeutic approach was implemented in vitro by applying the drug at different circadian times to synchronized LN-229 human GBM cells. Cell cultures displayed a functional circadian clock measured by Bmal1-luc activity and BMAL1 protein expression. Expression of Tiam1 was also assessed, finding circadian expression rhythms. The effects of 1A-116 on either cell proliferation, motility and toxicity, were dependent on the circadian cellular clock. The preliminary in vivo studies consisted on treating nude mice with 1A or control at ZT3 or 12. We found that the median survival of the mice treated at ZT3 was 82 days and at ZT12 was 78 days. These results suggest that a chronopharmacological approach based on the drug 1A-116 is a feasible strategy to improve GBM treatment outcomes. Citation Format: Laura Lucia Trebucq, Goergina A. Cardama, Pablo Lorenzano Menna, Diego A. Golombek, Juan J. Chiesa, Luciano Marpegan. The response of human glioblastoma cells to the novel drug 1A-116 is dependent on the circadian clock [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 620.
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We previously reported that early night peripheral bacterial lipopolysaccharide (LPS) injection produces phase delays in the circadian rhythm of locomotor activity in mice. We now assess the effects of proinflammatory cytokines on... more
We previously reported that early night peripheral bacterial lipopolysaccharide (LPS) injection produces phase delays in the circadian rhythm of locomotor activity in mice. We now assess the effects of proinflammatory cytokines on circadian physiology, including their role in LPS-induced phase shifts. First, we investigated whether differential systemic induction of classic proinflammatory cytokines could explain the time-specific behavioral effects of peripheral LPS. Induction levels for plasma interleukin (IL)-1α, IL-1β, IL-6, or tumor necrosis factor (TNF)-α did not differ between animals receiving a LPS challenge in the early day or early night. We next tested the in vivo effects of central proinflammatory cytokines on circadian physiology. We found that intracerebroventricular (i.c.v.) delivery of TNF-α or interleukin IL-1β induced phase delays on wheel-running activity rhythms. Furthermore, we analyzed if these cytokines mediate the LPS-induced phase shifts and found that i.c.v. administration of soluble TNF-α receptor (but not an IL-1β antagonistic) prior to LPS stimulation inhibited the phase delays. Our work suggests that the suprachiasmatic nucleus (SCN) responds to central proinflammatory cytokines in vivo, producing phase shifts in locomotor activity rhythms. Moreover, we show that the LPS-induced phase delays are mediated through the action of TNF-α at the central level, and that systemic induction of proinflammatory cytokines might be necessary, but not sufficient, for this behavioral outcome.
Research Interests: Chronobiology, Animal Behavior, Biology, Cytokines, Proinflammatory Cytokines, and 14 moreMedicine, Biological Sciences, Lipopolysaccharide, Mice, Circadian Rhythm, Animals, Male, Lipopolysaccharides, Circadian, Tumor necrosis factor-alpha, Suprachiasmatic Nucleus, Phase Shift, Motor activity, and Medical and Health Sciences
Research Interests: Endocrinology, Immunology, Animal Behavior, Biology, Medicine, and 15 moreNeuroimmunology, Lipopolysaccharide, Internal Medicine, Escherichia coli, Mice, Circadian Rhythm, Animals, Male, Body Temperature, Immune system, Lipopolysaccharides, Analysis of Variance, Circadian Clock, Neurosciences, and Motor activity
Research Interests: Neuroscience, Psychology, Biology, Cell Biology, Medicine, and 15 moreMice, Circadian Rhythm, Animals, Male, Astrocyte, Astrocytes, Fluorescent Antibody Technique, Immune system, Lipopolysaccharides, Analysis of Variance, Glial Fibrillary Acidic Protein, Circadian Clock, Neurosciences, Interleukin, and In Vitro Techniques
Many immune parameters exhibit daily and circadian oscillations, including the number of circulating cells and levels of cytokines in the blood. Mice also have a differential susceptibility to lipopolysaccharide (LPS or endotoxin)-induced... more
Many immune parameters exhibit daily and circadian oscillations, including the number of circulating cells and levels of cytokines in the blood. Mice also have a differential susceptibility to lipopolysaccharide (LPS or endotoxin)-induced endotoxic shock, depending on the administration time in the 24 h light-dark (LD) cycle. We replicated these results in LD, but we did not find temporal differences in LPS-induced mortality in constant darkness (DD). Animals challenged with LPS showed only transient effects on their wheel locomotor activity rhythm without modification of circadian period and phase. Levels of several key factors involved in the pathology of sepsis and septic shock were tested in LD. We found that LPS-induced levels of interleukin (IL)-1beta, IL-6, JE (MCP-1), and MIP1alpha were significantly higher at zeitgeber time (ZT) 11 (time of increased mortality) than at ZT19 (ZT12 = time of lights-off in the animal quarters for the 12L:12D condition). Our results indicate that the differences found in mortality that are dependent on the time of LPS-challenge are not directly related to an endogenous circadian clock, and that some relevant immune factors in the development of sepsis are highly induced at ZT11, the time of higher LPS-induced mortality, compared to ZT19.
Research Interests: Chronobiology, Biology, Cytokines, Medicine, Biological Sciences, and 15 moreLipopolysaccharide, Mice, Circadian Rhythm, Animals, Male, Immune system, Lipopolysaccharides, Circadian, Diurnal Variation, Photoperiod, Chemokines, Circadian Clock, Motor activity, Medical and Health Sciences, and Inflammation Mediators
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The Ras homologous family of small guanosine triphosphate-binding enzymes (GTPases) is critical for cell migration and proliferation. The novel drug 1A-116 blocks the interaction site of the Ras-related C3 botulinum toxin substrate 1... more
The Ras homologous family of small guanosine triphosphate-binding enzymes (GTPases) is critical for cell migration and proliferation. The novel drug 1A-116 blocks the interaction site of the Ras-related C3 botulinum toxin substrate 1 (RAC1) GTPase with some of its guanine exchange factors (GEFs), such as T-cell lymphoma invasion and metastasis 1 (TIAM1), inhibiting cell motility and proliferation. Knowledge of circadian regulation of targets can improve chemotherapy in glioblastoma. Thus, circadian regulation in the efficacy of 1A-116 was studied in LN229 human glioblastoma cells and tumor-bearing nude mice. Methods. Wild-type LN229 and BMAL1-deficient (i.e., lacking a functional circadian clock) LN229E1 cells were assessed for rhythms in TIAM1, BMAL1, and period circadian protein homolog 1 (PER1), as well as Tiam1, Bmal1, and Rac1 mRNA levels. The effects of 1A-116 on proliferation, apoptosis, and migration were then assessed upon applying the drug at different circadian times. Fin...
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Circadian disruption promotes tumor growth by changes in circadian rhythms of immune cells, clock genes, and cell cycle genes.
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Research Interests: Endocrinology, Circadian Rhythms, Medicine, Lipopolysaccharide, Hypothermia, and 11 moreInternal Medicine, Septic Shock, Circadian Rhythm, Biología, Sepsis, Ciencias Biológicas, Immune system, Tumor necrosis factor-alpha, Enseñanza - Aprendizaje Ciencias Naturales Y Exactas, Tumor Necrosis Factor–α (TNF), and Inflammatory response
Circadian systems enable organisms to synchronize their physiology to daily and seasonal environmental changes relying on endogenous pacemakers that oscillate with a period close to 24 h even in the absence of external timing cues. The... more
Circadian systems enable organisms to synchronize their physiology to daily and seasonal environmental changes relying on endogenous pacemakers that oscillate with a period close to 24 h even in the absence of external timing cues. The oscillations are achieved by intracellular transcriptional/translational feedback loops thoroughly characterized for many organisms, but still little is known about the presence and characteristics of circadian clocks in fungi other than Neurospora crassa. We sought to characterize the circadian system of a natural isolate of Aureobasidium pullulans, a cold-adapted yeast bearing great biotechnological potential. A. pullulans formed daily concentric rings that were synchronized by light/dark cycles and were also formed in constant darkness with a period of 24.5 h. Moreover, these rhythms were temperature compensated, as evidenced by experiments conducted at temperatures as low as 10 °C. Finally, the expression of clock-essential genes, frequency, white...
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The safety and efficacy of chemotherapeutics can vary as a function of the time of their delivery during the day. This study aimed to improve the treatment of glioblastoma (GBM), the most common brain cancer, by testing whether the... more
The safety and efficacy of chemotherapeutics can vary as a function of the time of their delivery during the day. This study aimed to improve the treatment of glioblastoma (GBM), the most common brain cancer, by testing whether the efficacy of the DNA alkylator temozolomide (TMZ) varies with the time of its administration. We found cell-intrinsic, daily rhythms in both human and mouse GBM cells. Circadian time of treatment affected TMZ sensitivity of murine GBM tumor cells in vitro. The maximum TMZ-induced DNA damage response, activation of apoptosis, and growth inhibition occurred near the daily peak in expression of the core clock gene Bmal1. Deletion of Bmal1 (Arntl) abolished circadian rhythms in gene expression and TMZ-induced activation of apoptosis and growth inhibition. These data indicate that tumor cell-intrinsic circadian rhythms are common to GBM tumors and can regulate TMZ cytotoxicity. Optimization of GBM treatment by timing TMZ administration to daily rhythms should b...
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Research Interests: Chronobiology, Endocrinology, Genetics, Mental Health, Metabolism, and 15 moreBiology, Cell Biology, Medicine, Earth and Environmental Sciences, Biological Sciences, Circadian Rhythm, Entrainment, Astrocytes, Ciencias Biológicas, Circadian, Glutamine Synthetase, Circadian Clock, Glutamate Uptake, Glutamate Receptor, and Medical and Health Sciences
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Research Interests: Neuroscience, Biology, Immunohistochemistry, Cell Biology, Medicine, and 14 moreClock, Cerebral Cortex, Mice, Circadian Rhythm, Animals, The, Astrocytes, Analysis of Variance, PER, Cryptochromes, Circadian Clock, Adenosine Triphosphate, Psychology and Cognitive Sciences, and Medical and Health Sciences
To interrogate endogenous p21 WAF1/CIP1 ( p21 ) promoter activity under basal conditions and in response to various forms of stress, knock-in imaging reporter mice in which expression of firefly luciferase ( FLuc ) was placed under the... more
To interrogate endogenous p21 WAF1/CIP1 ( p21 ) promoter activity under basal conditions and in response to various forms of stress, knock-in imaging reporter mice in which expression of firefly luciferase ( FLuc ) was placed under the control of the endogenous p21 promoter within the Cdkn1a gene locus were generated. Bioluminescence imaging (BLI) of p21 promoter activity was performed noninvasively and repetitively in mice and in cells derived from these mice. We demonstrated that expression of FLuc accurately reported endogenous p21 expression at baseline and under conditions of genotoxic stress and that photon flux correlated with mRNA abundance and, therefore, bioluminescence provided a direct readout of p21 promoter activity in vivo . BLI confirmed that p53 was required for activation of the p21 promoter in vivo in response to ionizing radiation. Interestingly, imaging of reporter cells demonstrated that p53 prevents the extracellular signal-regulated kinase/mitogen-activated p...
Research Interests: Molecular Biology, Biology, Cell Cycle, Medicine, Gene expression, and 14 moreWestern blotting, Biological Sciences, RNA interference, Molecular and cellular biology, Articles, Mice, Animals, Polymerase Chain Reaction, Bioluminescence, Luciferase, Hepatocytes, Gene Expression Regulation, Reporter gene, and Medical and Health Sciences
The hypothalamic suprachiasmatic nuclei (SCN), the site of a mammalian circadian clock, exhibit a dense immunoreactivity for glial fibrillary acidic protein (GFAP), a specific marker for astrocytes. Although there is evidence of a... more
The hypothalamic suprachiasmatic nuclei (SCN), the site of a mammalian circadian clock, exhibit a dense immunoreactivity for glial fibrillary acidic protein (GFAP), a specific marker for astrocytes. Although there is evidence of a circadian variation in GFAP‐IR in the hamster SCN and of the participation of glial cells in input and output mechanisms of the clock, the role of these cells within the circadian system is not clearly understood. The fact that astroglia can express and respond to cytokines suggests that they could work as mediators of immune signals to the circadian system. In the present study, we have found a daily variation of GFAP‐IR in the mouse SCN, peaking during the light phase. In addition, we have identified GFAP and nuclear factor‐κB (NF‐κB) in glial cells within the SCN and in primary cultures of the mouse SCN. Moreover, SCN glia cultures were transfected with an NF‐κB/luc construct whose transcriptional activity was increased with lipopolysaccharide 2 μg/ml, ...
Research Interests: Neuroscience, Psychology, Biology, Cell Biology, Medicine, and 15 moreMice, Circadian Rhythm, Animals, Male, Astrocyte, Astrocytes, Fluorescent Antibody Technique, Immune system, Lipopolysaccharides, Analysis of Variance, Glial Fibrillary Acidic Protein, Circadian Clock, Neurosciences, Interleukin, and In Vitro Techniques
Research Interests: Endocrinology, Immunology, Animal Behavior, Biology, Medicine, and 15 moreNeuroimmunology, Lipopolysaccharide, Internal Medicine, Escherichia coli, Mice, Circadian Rhythm, Animals, Male, Body Temperature, Immune system, Lipopolysaccharides, Analysis of Variance, Circadian Clock, Neurosciences, and Motor activity
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Many mammalian cell types show daily rhythms in gene expression driven by a circadian pacemaker. For example, cultured astrocytes display circadian rhythms in Period1 and Period2 expression. It is not known, however, how or which... more
Many mammalian cell types show daily rhythms in gene expression driven by a circadian pacemaker. For example, cultured astrocytes display circadian rhythms in Period1 and Period2 expression. It is not known, however, how or which intercellular factors synchronize and sustain rhythmicity in astrocytes. Because astrocytes are highly sensitive to vasoactive intestinal polypeptide (VIP), a neuropeptide released by neurons and important for the coordination of daily cycling, the authors hypothesized that VIP entrains circadian rhythms in astrocytes. They used astrocyte cultures derived from knock-in mice containing a bioluminescent reporter of PERIOD2 (PER2) protein, to assess the effects of VIP on the rhythmic properties of astrocytes. VIP induced a dose-dependent increase in the peak-to-trough amplitude of the ensemble rhythms of PER2 expression with maximal effects near 100 nM VIP and threshold values between 0.1 and 1 nM. VIP also induced dose- and phase-dependent shifts in PER2 rhyt...
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Research Interests:
Research Interests:
We previously reported that early night peripheral bacterial lipopolysaccharide (LPS) injection produces phase delays in the circadian rhythm of locomotor activity in mice. We now assess the effects of proinflammatory cytokines on... more
We previously reported that early night peripheral bacterial lipopolysaccharide (LPS) injection produces phase delays in the circadian rhythm of locomotor activity in mice. We now assess the effects of proinflammatory cytokines on circadian physiology, including their role in LPS-induced phase shifts. First, we investigated whether differential systemic induction of classic proinflammatory cytokines could explain the time-specific behavioral effects of peripheral LPS. Induction levels for plasma interleukin (IL)-1α, IL-1β, IL-6, or tumor necrosis factor (TNF)-α did not differ between animals receiving a LPS challenge in the early day or early night. We next tested the in vivo effects of central proinflammatory cytokines on circadian physiology. We found that intracerebroventricular (i.c.v.) delivery of TNF-α or interleukin IL-1β induced phase delays on wheel-running activity rhythms. Furthermore, we analyzed if these cytokines mediate the LPS-induced phase shifts and found that i.c.v. administration of soluble TNF-α receptor (but not an IL-1β antagonistic) prior to LPS stimulation inhibited the phase delays. Our work suggests that the suprachiasmatic nucleus (SCN) responds to central proinflammatory cytokines in vivo, producing phase shifts in locomotor activity rhythms. Moreover, we show that the LPS-induced phase delays are mediated through the action of TNF-α at the central level, and that systemic induction of proinflammatory cytokines might be necessary, but not sufficient, for this behavioral outcome.
Research Interests: Chronobiology, Animal Behavior, Biology, Cytokines, Proinflammatory Cytokines, and 14 moreMedicine, Biological Sciences, Lipopolysaccharide, Mice, Circadian Rhythm, Animals, Male, Lipopolysaccharides, Circadian, Tumor necrosis factor-alpha, Suprachiasmatic Nucleus, Phase Shift, Motor activity, and Medical and Health Sciences
Research Interests: Chronobiology, Endocrinology, Circadian Rhythms, Biology, Medicine, and 15 moreBiological Sciences, Lipopolysaccharide, Internal Medicine, Circadian Rhythm, Animals, Male, Light, Ciencias Biológicas, Cytokine, Lipopolysaccharides, PER, Biological clocks, Interleukin, Motor activity, and Medical and Health Sciences
Page 1. Biological Rhythm Research, 2000, Vol. 31, No. 1, pp. 56–70 0929-1016/00/3101-0056$15.00 © Swets & Zeitlinger Neurochemistry of Mammalian Entrainment: Signal Transduction Pathways in the Suprachiasmatic Nuclei ...
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Many immune parameters exhibit daily and circadian oscillations, including the number of circulating cells and levels of cytokines in the blood. Mice also have a differential susceptibility to lipopolysaccharide (LPS or endotoxin)-induced... more
Many immune parameters exhibit daily and circadian oscillations, including the number of circulating cells and levels of cytokines in the blood. Mice also have a differential susceptibility to lipopolysaccharide (LPS or endotoxin)-induced endotoxic shock, depending on the administration time in the 24 h light-dark (LD) cycle. We replicated these results in LD, but we did not find temporal differences in LPS-induced mortality in constant darkness (DD). Animals challenged with LPS showed only transient effects on their wheel locomotor activity rhythm without modification of circadian period and phase. Levels of several key factors involved in the pathology of sepsis and septic shock were tested in LD. We found that LPS-induced levels of interleukin (IL)-1beta, IL-6, JE (MCP-1), and MIP1alpha were significantly higher at zeitgeber time (ZT) 11 (time of increased mortality) than at ZT19 (ZT12 = time of lights-off in the animal quarters for the 12L:12D condition). Our results indicate that the differences found in mortality that are dependent on the time of LPS-challenge are not directly related to an endogenous circadian clock, and that some relevant immune factors in the development of sepsis are highly induced at ZT11, the time of higher LPS-induced mortality, compared to ZT19.