Copper Chelates Research Articles

Abstract 1418: Priming of the glioblastoma tumour microenvironment via copper chelation to enhance the efficacy of immunotherapies

Abstract Glioblastoma, a highly aggressive brain tumor with a dismal prognosis necessitates novel therapeutic approaches as current standard treatments yield limited success. Despite the transformative impact of immune checkpoint blockade in various malignancies, its efficacy in glioblastoma remains elusive. Similarly, chimeric antigen receptor T (CAR-T) cell therapy, while successful in hematological malignancies, faces challenges in solid tumors. This study addresses the immunosuppressive barriers hindering the effectiveness of immunotherapies in glioblastoma. Highlighting copper's pivotal role in glioblastoma, our research group demonstrated its elevation and influence on PD-L1 expression. Given the success of copper chelators in treating Wilson's Disease, in which neurological manifestations are alleviated following treatment, we hypothesized that copper chelation therapy could be a promising strategy for glioblastoma, especially when combined with immunotherapies. Flow cytometry analysis revealed a significant increase in the tumor-associated antigen GD2 following treatment with the copper chelator trientine (TETA), suggesting its potential synergy with GD2-targeted immunotherapies. In an immunocompetent murine glioblastoma model, combination therapy with GD2-targeted CAR-T cells and TETA effectively slowed tumor growth, demonstrating enhanced efficacy. Additionally, TETA as a monotherapy significantly improved survival compared to the control. The combination of TETA with anti-PD1 antibody therapy also exhibited promising results in inhibiting tumor growth and subsequently prolonging survival, with some mice demonstrating complete tumor regression. Importantly, TETA decreased copper levels in the brains of the mice, indicating the drug can act on the brain, a crucial therapeutic hurdle that is hampering the efficacy of novel therapies in glioblastoma. Further exploration of immune cell populations using OPAL Multiplex immunohistochemistry following TETA, anti-PD1, and GD2-targeted CAR-T cell treatments will underscore the potential of copper chelation in reshaping the tumor microenvironment. Our findings suggest that copper chelation, with agents like TETA, can be repurposed as a viable anticancer therapy when combined with immunotherapies. This study provides a compelling rationale for further pre-clinical validation, offering a potential breakthrough in the quest for effective glioblastoma treatments. Citation Format: Tyler Shai-Hee, Toni Rose Jue, Riccardo Cazzoli, Jourdin R. Rouaen, Filip Michniewicz, Jessica Bell, Tessa Gargett, Michael P. Brown, Orazio Vittorio. Priming of the glioblastoma tumour microenvironment via copper chelation to enhance the efficacy of immunotherapies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1418.

Abstract 3056: Copper chelation modulates YTHDF2 RNA-binding protein localization and global translation, in neuronal cancers

Abstract Cancers are dependent on copper for growth, angiogenesis and metastasis and copper chelators are in clinical trials against breast cancer, however the mechanisms of this emerging cancer treatment are still largely unexplored. We previously demonstrated that copper chelation has efficacy in mouse models of neuroblastoma and glioma. Multiomics analyses in neuronal cancers suggested that RNA-binding proteins were deregulated by copper chelator treatment. We found that the “RNA Binding” gene set was the most significantly ranked molecular function list using differentially expressed RNAs after copper chelation treatment (GO:0003723, p-value 1.02e−32). We subsequently found that the copper chelator drug decreased global protein synthesis, MYC/MYCN expression, as well as the amount a key methylated (m6A) RNA-binding protein called YTHDF2 in glioma cells, by RNA-seq (-log2FC 0.3, p-value 0.005) and western blot. This downregulation was also clear in neuroblastoma and glioblastoma cell lines. Interestingly, copper chelation also shifted YTHDF2 protein location into the nucleus. YTHDF2 knock-out (DepMAP Chronos dependency score, -0.32) and knock-down studies (siRNA pool, 38% decrease in cell confluence) in neuroblastoma cell lines and others, indicated that YTHDF2 depletion is selective against neuronal cancer cell growth. YTHDF2 was found to be a MYCN transcriptional target. Moreover, as YTHDF2 protein also binds MYCN transcripts (the principal oncogene in neuroblastoma), and copper signaling mRNAs, it appears that copper signaling, YTHDF2 and MYCs operates in an oncogenic-axis. Ongoing studies are investigating novel YTHDF2 inhibitors and their neuronal cancer efficacy. Our work represents a significant breakthrough in drugging oncogenic post-transcriptional processes either directly, or indirectly via depleting copper. Citation Format: Jessica L. Bell, Domenico K. Abruzzese, Erin Kasiou, Vu Vu Pham, Jean B. Bertoldo, Filip Michniewicz, Orazio Vittorio. Copper chelation modulates YTHDF2 RNA-binding protein localization and global translation, in neuronal cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3056.

Abstract 5788: Targeting p97-NPL4 pathway by copper chelators

Abstract Copper chelators forming stable complexes, such as the bis(diethyldithiocarbamate)-copper complex (CuET), derived from Antabuse (disulfiram, DSF), have shown pronounced anticancer activity. CuET induces aggregation of NPL4, a crucial cofactor of the p97 segregase in the ubiquitin-proteasome system, leading to proteotoxic stress and subsequent cancer cell death. Our study investigates whether NPL4 inhibition is unique to CuET or extends to other dithiocarbamate-copper complexes and distinct copper chelators. Utilizing a cellular NPL4 aggregation screening assay, we examined various chelators for their ability to induce NPL4 aggregation. Our findings reveal that this capacity is relatively common among structurally diverse DTCs-copper complexes and other unrelated copper chelators. Compounds that caused NPL4 aggregation also triggered NPL4-p97 complex immobilization, polyubiquitinated protein accumulation, and unfolded protein and heat shock responses. These responses correlated strongly with cancer cell cytotoxicity, particularly potent at nanomolar concentrations, underscoring NPL4 as a critical target. Our results demonstrate the broad potential of copper chelators in targeting NPL4, inducing lethal proteotoxic stress in cancer cells, and offering significant implications for drug development. Citation Format: Martin Mistrik, Zdenek Skrott, Martin Loffelmann. Targeting p97-NPL4 pathway by copper chelators [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5788.

Abstract 525: A comprehensive approach: Copper chelation therapy modulates epigenetic, kinase signaling and metabolic pathways in diffuse midline gliomas (DMG)

Abstract Diffuse Midline Gliomas (DMG) are incurable pediatric brain tumors driven by histone mutation H3K27M, globally reducing H3K27 trimethylation altering the epigenome and deregulating gene expression. These mutations often partner with and are strongly integrated into kinase signaling and metabolic alterations, driving aggressive tumor growth with many mechanisms which can compensate for one another and therefore evade traditional single agent therapies. Copper over-accumulation has been linked to epigenetic, kinase and metabolic dysregulation in various cancers and neurological disorders. Copper chelators are clinically approved for use in pediatric Wilson’s Disease patients, improve neurological symptoms and are under clinical investigation for several cancers. We propose copper chelators may simultaneously target these mechanisms in DMG. We performed transcriptomic interrogation of patient datasets, in vitro investigations in a panel of DMG cell lines and an in vivo investigation using an orthotopic xenograft model of DIPG (SU-DIPGVI-Luc). Copper chelator tetraethylenepentamine-pentachloride (TEPA) was used to treat DMG cells. Transcriptomic (RNA-Seq), proteomic, phospho-proteomic and metabolomic (Mass Spectrometry) investigations, western blots and immunofluorescent microscopy determined efficacy and mechanism. In vivo investigation consisted of survival study with concurrent luminescence imaging, histology, and copper assay. We demonstrate copper chaperone transcript expression is increased in DMG patient samples compared to non-DMG samples. In vitro, apoptosis and growth assays demonstrated H3K27M DMG were more sensitive to TEPA than H3-WT DMG. In H3K27M DMG, TEPA downregulated ATP, GTP and SAM metabolism which corresponded with downregulated kinase signaling and altered H3K27 trimethylation and DNA methylation patterns around cell cycle genes. Affected genes included those regulating centrosome segregation and ATP synthesis. Further transcriptomic and proteomic investigations highlighted TEPA downregulated G2-M phase kinase pathways, with immunofluorescence demonstrating TEPA reduced expression of mitotic marker p-H3S10 and epigenetic methylators EZH2 and DNMT1. In vivo, TEPA reduced brain copper accumulation, eliminated tumors and increased survival in an orthotopic patient-derived xenograft DMG model. Collectively, our data indicates TEPA induced multi-modal targeting of M-phase cell cycle progression through epigenetic, kinase signaling and metabolic mechanisms. Efficacy was observed both in vitro and in vivo, providing a promising basis for future investigation of copper chelators in DMG. We propose copper-chelation is a viable therapeutic strategy for DMG patients warranting clinical investigation. Citation Format: Filip Michniewicz, Riccardo Cazzoli, Jessica Bell, Federica Saletta, Ensieh Poursani, Jourdin R. Rouaen, Tyler Shai-Hee, Vu Pham, Toni Rose Jue, Matthew Biery, Liesl Bramberger, Terry Lim, Aaminah Khan, Federico M. Giorgi, Daniele Mercatelli, Chelsea Mayoh, Giuseppe Cirillo, Alexander Macmillan, Renee Whan, Christopher T. Barlow, Maria Tsoli, Nicholas Vitanza, Scott Berry, Pouya Faridi, David Ziegler, Orazio Vittorio. A comprehensive approach: Copper chelation therapy modulates epigenetic, kinase signaling and metabolic pathways in diffuse midline gliomas (DMG) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 525.

Abstract 2673: Copper chelation overcomes the immunosuppressive neuroblastoma microenvironment to potentiate anti-GD2 antibody therapy

Abstract Neuroblastoma is the most frequent extra-cranial pediatric cancer with high-risk disease possessing a dismal five-year survival rate <50%. Antibody-based therapies targeting the surface disialoganglioside GD2 have shown promising anti-tumor activity in patients however efficacy is strongly hampered by immunosuppressive mechanisms present in the tumor microenvironment. Neuroblastomas typically feature low mutational burdens, limited major histocompatibility complex class I expression, increased immune checkpoint markers, sparse immune infiltration, and soluble mediators which drastically limit anti-tumor immunity. Given the emerging link between elevated copper levels and tumoral immune evasion, we explored copper chelation as a therapeutic strategy using the agents tetraethylenepentamine (TEPA) and the clinically approved analogue triethylenetetramine (TETA). Using the immunocompetent TH-MYCN model, we performed single-cell RNA sequencing supported by OPAL multiplex immunohistochemistry and cytokine profiling of resected tumors to assess cellular and molecular changes occurring after one week of TEPA treatment. Therapy was observed to successfully deplete intratumoral copper and reinvigorate anti-tumor immunity with increased infiltration and activity of pro-inflammatory immune cells, and was specifically found to induce egress and N1-polarisation of neutrophils. Further mechanistic studies in vitro revealed the sequestration of copper by neuroblastoma cells attenuated neutrophil function, which could be successfully reversed upon TEPA treatment. Findings propose a novel mechanism of immune evasion, highlighting copper chelation as a therapeutic strategy to counteract immunosuppression. Using the TH-MYCN model, TEPA was employed as a priming agent to effectively potentiate anti-GD2 antibody therapy and achieve durable tumor control. Importantly, this was associated with increased Fc-receptor-bearing natural killer and myeloid cells, which elicit tumor clearance via antibody-dependent cellular cytotoxicity. Currently indicated for the treatment of copper overload disorder Wilson’s disease, TETA was evaluated for potential repurposing as a novel immunomodulatory agent. Using the syngeneic NXS2 model, TETA exhibited an exceptional safety profile and combination therapy achieved durable tumor control with no relapses occurring after treatment cessation. This result was similarly associated with the infiltration of pro-inflammatory immune cells, including N1 neutrophils. Collectively, this study credentials copper chelation as a non-toxic strategy to disrupt the immunosuppressive tumor microenvironment and reinvigorate anti-tumour immunity. Findings provide crucial translational evidence for repurposing TETA to potentiate anti-GD2 antibody therapy and improve neuroblastoma patient responses. Citation Format: Jourdin R. Rouaen, Antonella Salerno, Fabio Luciani, Jayne Murray, Tyler Shai-Hee, Nicodemus Tedla, Michelle Haber, Toby N. Trahair, Orazio Vittorio. Copper chelation overcomes the immunosuppressive neuroblastoma microenvironment to potentiate anti-GD2 antibody therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2673.

Copper and Copper Complexes in Tumor Therapy.

Copper (Cu), a crucial trace element in physiological processes, has garnered significant interest for its involvement in cancer progression and potential therapeutic applications. The regulation of cellular copper levels is essential for maintaining copper homeostasis, as imbalances can lead to toxicity and cell death. The development of drugs that target copper homeostasis has emerged as a promising strategy for anticancer treatment, with a particular focus on copper chelators, copper ionophores, and novel copper complexes. Recent research has also investigated the potential of copper complexes in cancer therapy.

Copper ion incorporation in α-synuclein amyloids.

Copper ion dys-homeostasis is linked to neurodegenerative diseases involving amyloid formation. Even if many amyloidogenic proteins can bind copper ions as monomers, little is known about copper interactions with the resulting amyloid fibers. Here, we investigate copper interactions with α-synuclein, the amyloid-forming protein in Parkinson's disease. Copper (Cu(II)) binds tightly to monomeric α-synuclein in vitro involving the N-terminal amine and the side chain of His50. Using purified protein and biophysical methods in vitro, we reveal that copper ions are readily incorporated into the formed amyloid fibers when present at the start of aggregation reactions, and the metal ions also bind if added to pre-formed amyloids. Efficient incorporation is observed for α-synuclein variants with perturbation of either one of the high-affinity monomer copper-binding residues (i.e., N-terminus or His50) whereas a variant with both N-terminal acetylation and His50 substituted with Ala does not incorporate any copper into the amyloids. Both the morphology of the resulting α-synuclein amyloids (amyloid fiber pitch, secondary structure, proteinase sensitivity) and the copper chemical properties (redox activity, chemical potential) are altered when copper is incorporated into amyloids. We speculate that copper chelation by α-synuclein amyloids contributes to the observed copper dys-homeostasis (e.g., reduced bioavailable levels) in Parkinson's disease patients. At the same time, amyloid-copper interactions may be protective to neuronal cells as they will shield aberrantly free copper ions from promotion of toxic reactive oxygen species.

Disulfiram Inhibits Opsonin-Independent Phagocytosis and Migration of Human Long-Lived In Vitro Cultured Phagocytes from Multiple Inflammatory Diseases.

Disulfiram (DSF), an anti-alcoholism medicine, exerts treatment effects in patients suffering from persistent Borreliosis and also exhibits anti-cancer effects through its copper chelating derivatives and induction of oxidative stress in mitochondria. Since chronic/persistent borreliosis is characterized by increased amounts of pro-inflammatory macrophages, this study investigated opsonin-independent phagocytosis, migration, and surface marker expression of in vivo activated and in vitro cultured human monocyte-derived phagocytes (macrophages and dendritic cells) with and without DSF treatment. Phagocytosis of non-opsonized Dynabeads® M-450 and migration of macrophages and dendritic cells were monitored using live cell analyzer Juli™ Br for 24 h, imaging every 3.5 min. To simultaneously monitor phagocyte function, results were analyzed by a newly developed software based on the differential phase contrast images of cells before and after ingestion of Dynabeads. DSF decreased the phagocytic capacities exhibited by in vitro enriched and long-lived phagocytes. Although no chemotactic gradient was applied to the test system, vigorous spontaneous migration was observed. We therefore set up an algorithm to monitor and quantify both phagocytosis and migration simultaneously. DSF not only reduced phagocytosis in a majority of these long-lived phagocytes but also impaired their migration. Despite these selective effects by DSF, we found that DSF reduced the expression densities of surface antigens CD45 and CD14 in all of our long-lived phagocytes. In cells with a high metabolic activity and high mitochondrial contents, DSF led to cell death corresponding to mitochondrial oxidative stress, whereas metabolically inactive phagocytes survived our DSF treatment protocol. In conclusion, DSF affects the viability of metabolically active phagocytes by inducing mitochondrial stress and secondly attenuates phagocytosis and migration in some long-lived phagocytes.

Simulated gastric leachate of 3D printer metal-fill filaments induces cytotoxic effects in rat and human intestinal models

Metals are used in 3-dimensional (3D) printer filaments in the manufacture of 3D printed objects. Exposure to the filaments, printed objects and emissions from printing may pose health risks from release of toxic metals. This study investigated the cytotoxicity of extruded 3D printer filament leachates in rat and human intestinal cells. Copper-, bronze-, and steel-fill extruded filaments were incubated in acidic media for 2 h. Leachates were adjusted to pH 7 and cells exposed for 4 or 24 h. Concentration- and time-dependent decreases in rat and human cell viability were observed using a colorimetric assay and confirmed by microscopy. Copper- and bronze-fill leachates were more cytotoxic than steel. Copper-fill leachates had the highest copper concentrations by ICP-MS. Exposure to CuSO4 resulted in concentration-dependent cytotoxicity in rat cells. The copper chelator bathocuproine disulphonate alleviated cytotoxicity of CuSO4 and copper-fill leachate, suggesting that copper ions have a role in the cytotoxicity. Hydrogen peroxide increased and glutathione decreased in rat cells exposed to copper-fill leachate, suggesting the formation of reactive oxygen species. Overall, our data indicate that metals released from the acidic exposure of print objects using metal-fill filaments, especially copper, are toxic to rat and human intestinal cells and additional studies are needed.

Effect of Sargassum polycystum on Expression of Tyrosinase and Production of Melanin

Introduction: Melanogenesis, the process responsible for producing melanin pigment, hinges on the activation of the amino acid L-tyrosine by the pivotal enzyme tyrosinase. In cases of hyperpigmentation induced by ultraviolet radiation, there's a surge in both tyrosinase enzyme levels and melanin production. Sargassum polycystum is currently under active investigation for its potential in this regard. This review aims to delve deeper into the properties of Sargassum polycystum and its impact on tyrosinase expression and melanin production. Literature Review: Sargassum polycystum, a brown seaweed commonly found in coastal regions with abundant coral reefs, harbors active compounds like flavonoids, fucoidan, fucoxanthin, and phloroglucinol. Fucoidan, an extract from brown seaweed, exhibits promising anti-inflammatory and antioxidant properties. Recent studies have revealed its ability to impede the melanogenesis process. Specifically, it was observed that fucoidan from brown seaweed extract could dampen α-MSH stimulation while enhancing the activation of the ERK-MAPK pathway, thereby acting as a negative regulator for melanogenesis. Fucoxanthin, another component, showcases antioxidant capabilities. Furthermore, phloroglucinol, found in Sargassum polycystum, exerts tyrosinase inhibitory effects by chelating copper within the enzyme. Flavonoids like catechin, hesperidin, kaempferol, and baicalein in Sargassum polycystum demonstrate inhibitory effects on melanogenesis activation pathways, including suppression of the Erk1/2 pathway, MiTF, and direct inhibition of tyrosinase. Consequently, this leads to reduced tyrosinase expression and melanin production, ultimately attenuating the melanogenesis process. Conclusion: Sargassum polycystum emerges as a promising candidate for combating hyperpigmentation induced by exposure to ultraviolet B radiation.

Enfermedad de Wilson: de la clínica a la genética

Wilson’s disease is a rare, autosomal recessive disorder caused by excess copper stored in the body. Mutations in the ATP7B gene are associated with Wilson’s Disease. It encodes the ATP7B protein responsible for maintaining normal levels of systemic copper. Intracellular copper overload leads to stress and subsequent oxidative damage to cellular proteins, lipids, DNA, RNA and mitochondria. Wilson’s disease has a wide clinical spectrum, mainly affecting the liver, nervous system, eye and mental health. The diagnosis requires clinical, biochemical, histological and genetic criteria. Treatment should be started early and if followed properly it can prevent clinical deterioration and improve life expectancy and quality of life. Treatment includes dietary measures, pharmacological treatments such as copper chelators and zinc salts, and in certain cases, liver transplantation. Some gene therapies are in the clinical trial phase and could become curative treatments for this disease.

Antioxidant and angiotensin converting enzyme inhibitory activities of red lionfish (Pterois volitans L.) muscle protein hydrolysates obtained using pepsin-pancreatin system

Despite being an invasive species and representing a threat in the area, red lionfish (Pterois volitans L.) meat is valued for its pleasant taste and high protein content. The objective of the present work was to evaluate the angiotensin converting enzyme inhibition (ACE-I) and the antioxidant activities of red lionfish muscle protein hydrolysates in vitro. Hydrolysates were obtained using the pepsin-pancreatin system, and their degree of hydrolysis (DH), electrophoretic, and amino acid profile were determined. Subsequently, their ACE-I and ABTS+ radical scavenging activity were evaluated, selecting those with the highest response. The highest DH (66.1%) was found in the hydrolysate obtained at 120 min of reaction time (H120); electrophoresis revealed the action of the enzymes on the muscle proteins. The highest bioactivities occurred during hydrolysis with pepsin for 60 min (H60), obtaining the highest ACE-I of 68.8% (evaluated with 500 µg protein) with a DH of 16.8%, ACE-I related amino acid content of 33.6%, and hydrophobic amino acid content of 42.9%. With the 30 min hydrolysates (H30), the highest Trolox equivalent antioxidant capacity of 60.3 mM/mg protein was obtained with 18.4% DH and 22.1% radical scavenger amino acids; its comparison with H120 proved that higher DH did not produce a greater response in ABTS+ radical scavenging activity, DPPH, reducing power, and copper chelation assays; with β-carotene bleaching being the exception. Therefore, it can be concluded that red lionfish muscle hydrolysates could be a promising source of peptides with antihypertensive and antioxidant properties, and can be suitable as functional ingredients.

Recurrent spontaneous pneumothorax in a patient with wilson disease who is on long-term D-penicillamine: Case report

Introduction: Recurrent spontaneous pneumothoraces (PSP) in association with liver disease is well known to occur with alpha 1 antitrypsin deficiency (AATD). Wilson disease (WD) association with recurrent PSP is not a known entity. Case presentation: A 42-year-old nonsmoker man with a history of recurrent PSP was admitted with a large right pneumothorax. Following intercostal chest drain (ICD) patient’s breathing improved and the lung expanded. Despite that, he continued to have a small residual pneumothorax (<2cm). Twelve years ago, he was diagnosed with WD. The patient was on treatment with D-penicillamine. After 3 years of the diagnosis of WD, the patient developed left-sided recurrent PSPs where he ended up with a left-pleurectomy in 2015. For the current event, initially expectant management was done for residual pneumothorax. All the possible causes for PSP were excluded. On follow-up, he continued to be symptomatic. A thoracic surgical referral was arranged for Video-Assisted Thoracoscopic Surgery (VATS) and redo-pleurectomy. The safety of future treatment with D-penicillamine is not concluded as there is no convincing evidence to prove it as a culprit agent for PSPs. Multidisciplinary discussion was arranged and consideration of treatment with alternative copper chelating therapy was emphasized. Conclusion: Though rare, it is important to observe for occurrence of PSP in WD patients. This case report will be eye-opening for the association of lung disease and WD and related treatment.

Meat productivity and quality of meat and liver of broiler chickens after consuming diet with mineral supplements containing chelates of cuprum, iron, zinc, and manganese

Poultry farming plays an important role as a producer of high-quality animal protein, which can solve issues of protein deficit among the population. Genetic growth potential of broiler chickens can be maximized through regular supply of microelements, especially chelate aminoacids. Those compounds improve metabolic processes in the body and increase increments in live mass and slaughter-yield of meat. The main objective of the study was identifying effects of mineral supplements with chelate microminerals on meat productivity of broiler chickens, quality of breast and thigh muscles, and content of mineral compounds in meat and liver. The studies were carried out on Kobb-500 broiler chickens, divided into three groups, each comprising 25 individuals. The experiment lasted from day 1 to day 42 of raising poultry. Broilers of the control group received the basic diet balanced in nutrients; the diet of birds of the second group was supplemented with Kronotsyd-L with chelates of copper, iron, zinc, and manganese in calculation of 0.25 mL/L of water, and chickens of the third consumed a diet with copper glycinate in calculation of 0.30 mL/L of water. Subject to mineral supplements with chelate microelements, the chickens had the highest mean-daily increments of live mass, slaughter yield, and yield of breast muscles. Use of mineral supplements in the diet of broiler chickens affected the quality of their meat, in particular its chemical composition. Those supplements led to changes in mineral composition of meat, influencing the concentrations of individual minerals in the tissues of the birds. Since minerals are important components of protein metabolism, the supplements augmented the content of dry matter and protein in meat. In the broiler chickens that had been receiving copper glycinate, the breast muscles had statistically significant increases in the mass shares of dry matter and protein. We found decrease in the content of fat in the breast and thigh muscles and the liver subject to copper glycinate and Kronotsyd-L with chelates of copper, iron, zinc, and manganese. This indicates improvement of the quality of meat and the liver. Use of mineral supplements with chelated microelements increased the ash content in dry matter because of higher concentrations of phosphorus, calcium, magnesium, iron, and manganese.

Erinacine S, a small active component derived from Hericium erinaceus, protects oligodendrocytes and alleviates mood abnormalities in cuprizone-exposed rodents

Hericium erinaceus mycelium extract (HEM), containing erinacine A (HeA) and erinacine S (HeS), has shown promise in promoting the differentiation of oligodendrocyte precursor cells (OPCs) into mature oligodendrocytes (OLs), crucial for myelin production in the central nervous system (CNS). The main aim of this study was to characterize the protective effects of HEM and its components on OLs and myelin in demyelinating rodents by exposure to cuprizone (CPZ), a copper chelating agent commonly used to induce demyelination in the corpus callosum of the brain. Rats were fed by CPZ-containing diet and simultaneously orally administered HEM, HeA, or HeS on a daily basis for three weeks. We found that HEM and HeS preserved myelin and OLs in the corpus callosum of CPZ-fed rats, along with reduced microglia and astrocyte activation, and downregulated IL-1β expression. Furthermore, post-treatment with HeS, in mouse models with acute (6 weeks) or chronic (12 weeks) CPZ-induced demyelination demonstrated oral administration during the final 4 weeks (HeS4/6 or HeS4/12) effectively preserved myelin in the corpus callosum. Additionally, HeS4/6 and HeS4/12 inhibited anxious and depressive-like behaviors in CPZ-fed mice. In summary, simultaneous administration of HEM and HeS in rats during short-term CPZ intoxication preserved OLs and myelin. Furthermore, post-administration of HeS not only inhibited demyelination and gliosis but also alleviated anxiety and depression in both acute and chronic CPZ-fed mice. This study presents compelling evidence supporting the potential of HeS as a promising small active compound for protecting OLs and preserving myelin in demyelinating diseases associated with emotional disorders.

Copper Promotes LPS-Induced Inflammation via the NF-кB Pathway in Bovine Macrophages.

Inflammation is a complex physiological process that enables the clearance of pathogens and repairing damaged tissues. Elevated serum copper concentration has been reported in cases of inflammation, but the role of copper in inflammatory responses remains unclear. This study used bovine macrophages to establish lipopolysaccharide (LPS)-induced inflammation model. There were five groups in the study: a group treated with LPS (100ng/ml), a group treated with either copper chelator (tetrathiomolybdate, TTM) (20μmol) or CuSO4 (25μmol or 50μmol) after LPS stimulation, and a control group. Copper concentrations increased in macrophages after the LPS treatment. TTM decreased mRNA expression of pro-inflammatory factors (IL-1β, TNF-α, IL-6, iNOS, and COX-2), whereas copper supplement increased them. Compared to the control group, TLP4 and MyD88 protein levels were increased in the TTM and copper groups. However, TTM treatment decreased p-p65 and increased IкB-α while the copper supplement showed reversed results. In addition, the phagocytosis and migration of bovine macrophages decreased in the TTM treatment group while increased in the copper treatment groups. Results mentioned above indicated that copper could promote the LPS-induced inflammatory response in bovine macrophages, promote pro-inflammatory factors by activating the NF-кB pathway, and increase phagocytosis capacity and migration. Our study provides a possible targeted therapy for bovine inflammation.

NIR-IIb fluorescence antiangiogenesis copper nano-reaper for enhanced synergistic cancer therapy.

The formation of blood vessel system under a relatively higher Cu2+ ion level is an indispensable precondition for tumor proliferation and migration, which was assisted in forming the tumor immune microenvironment. Herein, a copper ions nano-reaper (LMDFP) is rationally designed not only for chelating copper ions in tumors, but also for combination with photothermal therapy (PTT) to improve antitumor efficiency. Under 808nm laser irradiation, the fabricated nano-reaper converts light energy into thermal energy to kill tumor cells and promotes the release of D-penicillamine (DPA) in LMDFP. Photothermal properties of LMDFP can cause tumor ablation in situ, which further induces immunogenic cell death (ICD) to promote systematic antitumor immunity. The released DPA exerts an anti-angiogenesis effect on the tumor through chelating copper ions, and inhibits the expression of programmed death ligand 1 (PD-L1), which synergizes with PTT to enhance antitumor immunity and inhibit tumor metastasis. Meanwhile, the nanoplatform can emit near-infrared-IIb (NIR-IIb) fluorescence under 980nm excitation, which can be used to track the nano-reaper and determine the optimal time point for PTT. Thus, the fabricated nano-reaper shows powerful potential in inhibiting tumor growth and metastasis, and holds great promise for the application of copper nanochelator in precise tumor treatment.

Microglial Neuroinflammation-Independent Reversal of Demyelination of Corpus Callosum by Arsenic in a Cuprizone-Induced Demyelinating Mouse Model.

Demyelination is the loss of myelin in CNS, resulting in damaged myelin sheath. Oxidative stress and neuroinflammation play a key role in inducing demyelinating diseases like MS; hence, controlling oxidative stress and neuroinflammation is important. Cuprizone (CPZ), a copper chelator, generates oxidative stress and neuroinflammation, thereby inducing demyelination. Therefore, the CPZ-induced demyelinating mouse model (CPZ model) is widely used in research. The present study was intended to unravel a mechanism of inhibition of demyelination by arsenic in a CPZ model, which is otherwise known for its toxicity. We investigated an alternative mechanism of inhibition of demyelination by arsenic through the reversal of SOD1 activity employing in silico analysis, analytical chemistry techniques, and in vitro and in vivo experiments. In vivo experiments showed protection of body weight, survivability, and myelination of the corpus callosum in CPZ and arsenic-co-exposed animals, where neuroinflammation was apparently not involved. In vitro experiments revealed that arsenic-mediated reversal of impaired SOD1 activity leads to reduced cellular ROS levels and better viability of primary oligodendrocytes. Reversal of SOD1 activity was also observed in the corpus callosum tissue isolated from experimental animals. In silico and analytical chemistry studies revealed that similar to copper, arsenic can potentially bind to CPZ and thereby make the copper freely available for SOD1 activity. Suitable neurobehavior tests further validated the protective effect of arsenic. Taken together, the present study revealed that arsenic protects oligodendrocytes and demyelination of corpus callosum by reversing CPZ-induced impaired SOD1 activity.

Liver and mucous secretion enzymatic biomarkers of Eobania vermiculata treated with some newly synthesized acrylamide derivatives

Abstract Background Acrylamide derivatives have a potential biological activity as well as acting as precursors in many organic syntheses. Moreover, acrylamides and their derivatives cause convulsions and diffused damage to different sections of the nervous system of infected animals. Novel copper and zinc chelates originated from (E)-3-(4-bromophenyl)-2-cyanoacrylamide (L1), and (E)-2-cyano-3-(4-nitrophenyl)acrylamide (L2) were prepared, and their chemical skeletons were identified by infrared and mass spectra. The obtained compounds were screened in vitro against the brown garden snail, Eobania vermiculata using the contact method along 72 h. Stock solutions of tested compounds were prepared utilizing distilled water and DMSO mixture, and four concentrations of each compound were prepared (50, 150, 250 and 350 ppm). Eobania vermiculata snails were treated with LC50 concentrations of prepared compounds for 3 days, and live snails were used to estimate the level of some liver and mucous secretion enzymatic biomarkers: transaminases enzymes (Alanine Aminotransferase (ALT) and Aspartate Aminotransferase (AST)), Total Protein content (TP), Acid Phosphatases (ACP) and Alkaline Phosphatase (ALP). Results The results demonstrated that the examined compounds have a relatively toxic effect toward the screened species. Zinc complexes displayed a higher toxicity than copper ones. The results authenticated considerable high effects of the synthesized compounds on investigated enzymes. Conclusions The promising effects of Cu(II), Zn(II) complexes (1, 2) on stimulating the mucous secretion of tested snails are clear through the elevated levels of ALP and ACP enzymes of treated snails. The enhancement or reduction of AST, ALT level and TP content of treated snails demonstrated the effects of prepared compounds on liver functions of these species.

Cuproptosis: Unraveling the Mechanisms of Copper-Induced Cell Death and Its Implication in Cancer Therapy

Copper, an essential element for various biological processes, demands precise regulation to avert detrimental health effects and potential cell toxicity. This paper explores the mechanisms of copper-induced cell death, known as cuproptosis, and its potential health and disease implications, including cancer therapy. Copper ionophores, such as elesclomol and disulfiram, increase intracellular copper levels. This elevation triggers oxidative stress and subsequent cell death, offering potential implications in cancer therapy. Additionally, copper ionophores disrupt mitochondrial respiration and protein lipoylation, further contributing to copper toxicity and cell death. Potential targets and biomarkers are identified, as copper can be targeted to those proteins to trigger cuproptosis. The role of copper in different cancers is discussed to understand targeted cancer therapies using copper nanomaterials, copper ionophores, and copper chelators. Furthermore, the role of copper is explored through diseases such as Wilson and Menkes disease to understand the physiological mechanisms of copper. Exploring cuproptosis presents an opportunity to improve treatments for copper-related disorders and various cancers, with the potential to bring significant advancements to modern medicine.