The NLRP3 inflammasome: contributions to inflammation‑related diseases

The NOD-like receptor protein 3 (NLRP3) inflammasome is a protein complex that regulates innate immune responses by activating caspase-1 and the inflammatory cytokines interleukin (IL)-1β and IL-18. Multiple studies have demonstrated the importance of the NLRP3 inflammasome in the development of immune and inflammation-related diseases, including arthritis, Alzheimer’s disease, inflammatory bowel disease, and other autoimmune and autoinflammatory diseases. This review first explains the activation and regulatory mechanism of the NLRP3 inflammasome. Secondly, we focus on the role of the NLRP3 inflammasome in various inflammation-related diseases. Finally, we look forward to new methods for targeting the NLRP3 inflammasome to treat inflammation-related diseases, and provide new ideas for clinical treatment.National Natural Science Foundation of China, 82200330, 82070422, 81871607Youth Science and Technology Rising Star Project of Shaanxi Province2020KJXX-036, Innovation Capability Strong Foundation Plan of Xi’an City, 21YXYJ003

Melatonin Treatment Triggers Metabolic and Intracellular pH Imbalance in Glioblastoma

Supplementary Materials The following supporting information can be downloaded at: https://www.mdpi.com/article/10.3390/cells11213467/s1, Figure S1: Metabolic baseline characterization, treatments with melatonin agonists, and combina-tion treatments of melatonin (aMT) and the standard of care for GBM; Figure S2: Seahorse com-plementary analyses; Figure S3: pH fluctuations; Figure S4: In vivo results after melatonin intra-tumoral treatment.Funding This research was funded by the Distinguished Mayo Clinic Investigator Award (A.Q.-H.) and the William J. and Charles H. Mayo Professor (A.Q.-H.); the Uihlein Neuro-oncology Research Fund (A.Q.-H.) and R01CA200399 (A.Q.-H.).Metabolic rewiring in glioblastoma (GBM) is linked to intra- and extracellular pH regulation. In this study, we sought to characterize the role of melatonin on intracellular pH modulation and metabolic consequences to identify the mechanisms of action underlying melatonin oncostatic effects on GBM tumor initiating cells. GBM tumor initiating cells were treated at different times with melatonin (1.5 and 3.0 mM).We analyzed melatonin’s functional effects on GBM proliferation, cell cycle, viability, stemness, and chemo-radiosensitivity. We then assessed the effects of melatonin on GBM metabolism by analyzing the mitochondrial and glycolytic parameters. We also measured the intracellular and extracellular pH. Finally, we tested the effects of melatonin on a mouse subcutaneous xenograft model. We found that melatonin downregulated LDHA and MCT4, decreasing lactate production and inducing a decrease in intracellular pH that was associated with an increase in ROS and ATP depletion. These changes blocked cell cycle progression and induced cellular death and we observed similar results in vivo. Melatonin’s cytotoxic effects on GBM were due, at least in part, to intracellular pH modulation, which has emerged as a newly identified mechanism, providing new insights into the oncostatic effect of melatonin on GBM.Distinguished Mayo Clinic Investigator Award (A.Q.-H.)William J. and Charles H. Mayo Professor (A.Q.-H.)Uihlein Neuro-oncology Research Fund (A.Q.-H.)R01CA200399 (A.Q.-H.

Oncostatic effect of melatonin in head and neck cancer cells: clonogenic assay

Background: The oncostatic effect of melatonin has been previously described among different neoplastic types. One of these is head and neck squamous cell cancer (HNSCC) with a high rate of mortality and morbidity. Melatonin (aMT) could cause cell death in this neoplastic cell type. To determine this, we performed a clonogenic assay with CAL-27 cells treated with melatonin and/or radiation. Methods: Cells were plated in a 6-well plate, with 800 cells per well. Assays were performed at least twice and each time in triplicates. Cells were allowed to grow 15 days to form colonies of at least 50 cells each one. Cells were treated with melatonin (100, 500, 1000, 1500 and 2000 µM) alone or in combination with irradiation (8 Gy). To visualize colonies, cells were fixed in 100 % ethanol on days 12, 13, 14 and 15 after they were plated and stained with crystal violet solution. Colonies were scored with Image J Software. Results: The results clearly show that melatonin inhibits colony growth of CAL-27 cells in a dosedependent manner in the groups treated with melatonin alone 1500 µM or in combination with irradiation. Conclusion: The results show the capability of aMT to prevent colony growth and causing cell death on CAL-27 cancer cells, especially when combined with radiation. This is consistent with previous studies on aMT oncostatic effects and suggests that usage of aMT in vivo should have future clinical application

Composición de melatonina o sus derivados con coenzima q10 y su uso contra el envejecimiento de la piel

Número de publicación: ES2394245 B2 . Número de solicitud: 201231849.Composición de melatonina o sus derivados con coenzima Q10 Y su uso contra el envejecimiento de la piel. La presente invención se refiere a una composición que comprende melatonina, metabolito o derivado de la misma y coenzima Q10 Y su uso para la elaboración de una composición cosmética o farmacéutica para el tratamiento de la piel, ya que esta composición potencia la entrada de ambas moléculas en la mitocondria y facilita una absorción transdérmica, pudiendo alcanzar tanto la melatonina como la CoQ1O todos los estratos de la piel.Universidad de Granad

iMS-Bmal1−/− mice show evident signs of sarcopenia that are counteracted by exercise and melatonin therapies

This study was partially supported by grants from the Instituto de Salud Carlos III through the grants PI19‐01372 and CB/10/00238 (co‐funded by European Regional Development Fund/European Social Fund “Investing in your future”); the Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía (CTS‐101), Spain. José Fernández‐Martínez is supported by an FPU fellowship from the Ministerio de Educación, Spain; Yolanda Ramírez‐Casas has a PFIS fellowship from the Instituto de Salud Carlos III; Paula Aranda‐Martínez has a fellowship from grant no. P18‐RT‐698, and Alba López‐Rodríguez has a fellowship from grant no. P18‐RT‐3222, from the Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucía.Sarcopenia is an age-related disease characterized by a reduction in muscle mass, strength, and function and, therefore, a deterioration in skeletal muscle health and frailty. Although the cause of sarcopenia is still unknown and, thus, there is no treatment, increasing evidence suggests that chronodisruption, particularly alterations in Bmal1 clock gene, can lead to those deficits culminating in sarcopenia. To gain insight into the cause and mechanism of sarcopenia and the protective effect of a therapeutic intervention with exercise and/or melatonin, the gastrocnemius muscles of male and female skeletal muscle-specific and inducible Bmal1 knockout mice (iMS-Bmal1−/−) were examined by phenotypic tests and light and electron microscopy. Our results revealed a disruption of the normal activity/rest rhythm, a drop in skeletal muscle function and mass, and increased frailty in male and female iMS-Bmal1−/− animals compared to controls. A reduction in muscle fiber size and increased collagenous tissue were also detected, accompanied by reduced mitochondrial oxidative capacity and a compensatory shift towards a more oxidative fiber type. Electron microscopy further supports mitochondrial impairment in mutant mice. Melatonin and exercise ameliorated the damage caused by loss of Bmal1 in mutant mice, except for mitochondrial damage, which was worsened by the latter. Thus, iMS-Bmal1−/− mice let us to identify Bmal1 deficiency as the responsible for the appearance of sarcopenia in the gastrocnemius muscle. Moreover, the results support the exercise and melatonin as therapeutic tools to counteract sarcopenia, by a mechanism that does not require the presence of Bmal1.Ministerio de Educación, Spain P18‐RT‐3222, P18‐RT‐698Consejería de Economía, Innovación, Ciencia y Empleo, Junta de Andalucíanstituto de Salud Carlos III: CB/10/00238, PI19‐01372 ISCIIIEuropean Regional Development Fund ERDFJunta de Andalucía CTS‐10

From Chronodisruption to Sarcopenia: The Therapeutic Potential of Melatonin

Sarcopenia is an age-related condition that involves a progressive decline inmusclemass and function, leading to increased risk of falls, frailty, andmortality. Although the exactmechanisms are not fully understood, aging-related processes like inflammation, oxidative stress, reduced mitochondrial capacity, and cell apoptosis contribute to this decline. Disruption of the circadian system with age may initiate these pathways in skeletal muscle, preceding the onset of sarcopenia. At present, there is no pharmacological treatment for sarcopenia, only resistance exercise and proper nutrition may delay its onset. Melatonin, derived from tryptophan, emerges as an exceptional candidate for treating sarcopenia due to its chronobiotic, antioxidant, and anti-inflammatory properties. Its impact on mitochondria and organelle, where it is synthesized and crucial in aging skeletal muscle, further highlights its potential. In this review, we discuss the influence of clock genes in muscular aging, with special reference to peripheral clock genes in the skeletal muscle, as well as their relationship with melatonin, which is proposed as a potential therapy against sarcopenia.Instituto de Salud Carlos III, grant numbers PI19-01372 and CB/10/00238FPU fellowship from the Ministerio de Universidades, SpainPFIS fellowship from the Instituto de Salud Carlos II

Review Insights into the Role of Plasmatic and Exosomal microRNAs in Oxidative Stress-Related Metabolic Diseases

A common denominator of metabolic diseases, including type 2 diabetes Mellitus, dyslipidemia, and atherosclerosis, are elevated oxidative stress and chronic inflammation. These complex, multi-factorial diseases are caused by the detrimental interaction between the individual genetic background and multiple environmental stimuli. The cells, including the endothelial ones, acquire a preactivated phenotype and metabolic memory, exhibiting increased oxidative stress, inflammatory gene expression, endothelial vascular activation, and prothrombotic events, leading to vascular complications. There are different pathways involved in the pathogenesis of metabolic diseases, and increased knowledge suggests a role of the activation of the NF-kB pathway and NLRP3 inflammasome as key mediators of metabolic inflammation. Epigenetic-wide associated studies provide new insight into the role of microRNAs in the phenomenon of metabolic memory and the development consequences of vessel damage. In this review, we will focus on the microRNAs related to the control of anti-oxidative enzymes, as well as microRNAs related to the control of mitochondrial functions and inflammation. The objective is the search for new therapeutic targets to improve the functioning of mitochondria and reduce oxidative stress and inflammation, despite the acquired metabolic memoryCIBER-Consorcio Centro de Investigación Biomédica en Red-(CB16/10/00238Instituto de Salud Carlos III, Ministerio de Ciencia e InnovaciónGrant of the FUNDACIÓN EUGENIO RODRIGUEZ PASCUAL (ERP-2021),Call 2021, Spai

Role of c-miR-21, c-miR-126, Redox Status, and Inflammatory Conditions as Potential Predictors of Vascular Damage in T2DM Patients

The development of type 2 diabetes mellitus (T2DM) vascular complications (VCs) is associated with oxidative stress and chronic inflammation and can result in endothelial dysfunctions. Circulating microRNAs play an important role in epigenetic regulation of the etiology of T2DM. We studied 30 healthy volunteers, 26 T2DM patients with no complications, and 26 T2DM patients with VCs, to look for new biomarkers indicating a risk of developing VCs in T2DM patients. Peripheral blood samples were used to determine redox state, by measuring the endogenous antioxidant defense system (superoxide dismutase, SOD; catalase, CAT; glutathione reductase, GRd; glutathione peroxidase, GPx; and glucose-6-phosphate dehydrogenase, G6DP) and markers of oxidative damage (advanced oxidation protein products, AOPP; lipid peroxidation, LPO). Additionally, inflammatory marker levels (IL-1, IL-6, IL-18, and TNF- ), c-miR-21, and c-miR-126 expression were analyzed. T2DM patients showed the highest oxidative damage with increased GSSG/GSH ratios, LPO, and AOPP levels. In both diabetic groups, we found that diminished SOD activity was accompanied by increased CAT and decreased GRd and G6PD activities. Diabetic patients presented with increased relative expression of c-miR-21 and decreased relative expression of c-miR-126. Overall, c-miR-21, SOD, CAT, and IL-6 had high predictive values for diabetes diagnoses. Finally, our data demonstrated that IL-6 exhibited predictive value for VC development in the studied population. Moreover, c-miR- 21 and c-miR-126, along with GPx and AOPP levels, should be considered possible markers for VC development in future studies.University of GranadaEugenio Rodriguez Pascual Foundation ERP-2021CIBERfes (ISCIII, Spain) CB16-10-0023

Patient-derived tumor models in cancer research: Evaluation of the oncostatic effects of melatonin

MCIN/AEI/10.13039/ 501100011033/The European Regional Development Fund (B-CTS-071-UGR18)University of Granada (Grant “UNETE,” UCEPP2017- 05), SpainUnión Europea “NextGenerationEU”/ PRTR (PID2020-115112RB- I00; SAF2017-85903-P)Consejería de Economía , Innovación, Ciencia y Empleo, Junta de Andalucía (P18- RT-32222)Ministerio de Educación Cultura y Deporte, Spai

Understanding the Mechanism of Action of Melatonin, Which Induces ROS Production in Cancer Cells

Reactive oxygen species (ROS) constitute a group of highly reactive molecules that have evolved as regulators of important signaling pathways. In this context, tumor cells have an altered redox balance compared to normal cells, which can be targeted as an antitumoral therapy by ROS levels and by decreasing the capacity of the antioxidant system, leading to programmed cell death. Melatonin is of particular importance in the development of innovative cancer treatments due to its oncostatic impact and lack of adverse effects. Despite being widely recognized as a pro-oxidant molecule in tumor cells, the mechanism of action of melatonin remains unclear, which has hindered its use in clinical treatments. The current review aims to describe and clarify the proposed mechanism of action of melatonin inducing ROS production in cancer cells in order to propose future anti-neoplastic clinical applications.MCIN/AEI, SpainEuropean Commission SAF2017-85903-P ID2020-115112RB-I00Consejeria de Innovacion, Ciencia y Empresa Junta de Andalucia P07-CTS-03135 P10-CTS-5784 CTS-101University of Granada, Spain UCE-PP2017-05Spanish Governmen