Copper Depletion Research Articles

A Metal Chelation Therapy to Effectively Eliminate Breast Cancer and Intratumor Bacteria While Suppressing Tumor Metastasis by Copper Depletion and Zinc Ions Surge.

The intratumor microbiota results in the immunosuppressive microenvironment and facilitates tumor growth and metastasis. However, developing a synergistic therapy with antitumor, antibacterial, and antimetastatic effects faces enormous challenges. Here, we propose anmetal chelation therapy to effectively eliminate tumor and intratumor bacteria and suppress tumor metastasis. Different from traditional chelation therapy that only consumes metal elements, this therapy not only eliminates the crucial metal elements for tumor metabolism but also releases new metal ions with antitumor and antibacterial properties. Based on the high demand for copper in breast cancer, we prepare a fibrous therapeutic nanoagent (Zn-PEN) by chelatingD-Penicillamine (D-PEN) with Zn2+. Firstly, Zn-PEN achieves dual inhibition of oxidative phosphorylationand glycolysis metabolism through copper depletion and Zn2+ activated cGAS-STING pathway, thus inducing tumor cell death. Secondly, Zn-PEN has the capability to eradicate Fusobacterium nucleatumin breast cancer, thereby mitigating its immunosuppressive impact on the tumor microenvironment. Finally, Zn-PEN effectively inhibits tumor metastasis through multiple routes, including the inhibition of epithelial-mesenchymal transition process, activation of cGAS-STING pathway, and elimination with F. nucleatum. Therefore, we verify the feasibility of Zn-PEN mediated metal chelation therapy in a 4T1 model infected with F. nucleatum, providing a new therapeutic strategy for inhibiting tumor metastasis.

A Metal Chelation Therapy to Effectively Eliminate Breast Cancer and Intratumor Bacteria While Suppressing Tumor Metastasis by Copper Depletion and Zinc Ions Surge

The intratumor microbiota results in the immunosuppressive microenvironment and facilitates tumor growth and metastasis. However, developing a synergistic therapy with antitumor, antibacterial, and antimetastatic effects faces enormous challenges. Here, we propose an metal chelation therapy to effectively eliminate tumor and intratumor bacteria and suppress tumor metastasis. Different from traditional chelation therapy that only consumes metal elements, this therapy not only eliminates the crucial metal elements for tumor metabolism but also releases new metal ions with antitumor and antibacterial properties. Based on the high demand for copper in breast cancer, we prepare a fibrous therapeutic nanoagent (Zn‐PEN) by chelating D‐Penicillamine (D‐PEN) with Zn2+. Firstly, Zn‐PEN achieves dual inhibition of oxidative phosphorylation and glycolysis metabolism through copper depletion and Zn2+ activated cGAS‐STING pathway, thus inducing tumor cell death. Secondly, Zn‐PEN has the capability to eradicate Fusobacterium nucleatum in breast cancer, thereby mitigating its immunosuppressive impact on the tumor microenvironment. Finally, Zn‐PEN effectively inhibits tumor metastasis through multiple routes, including the inhibition of epithelial‐mesenchymal transition process, activation of cGAS‐STING pathway, and elimination with F. nucleatum. Therefore, we verify the feasibility of Zn‐PEN mediated metal chelation therapy in a 4T1 model infected with F. nucleatum, providing a new therapeutic strategy for inhibiting tumor metastasis.

Effect of solution‐processed cesium carbonate on Cu(In,Ga)Se2 thin‐film solar cells

AbstractHeavy alkali‐metal treatments have been the most recent breakthrough in improving the efficiency of Cu(In,Ga)Se2 (CIGS) solar cells. Alkali halides are generally evaporated onto the surface of the CIGS thin film by a vacuum process. Here, we report an alternative, low‐cost solution process for the surface treatment of CIGS thin films using cesium carbonate (CsCO3) as a new route to incorporate cesium (Cs) for improving solar cell performance. CIGS thin films were fabricated using pulsed hybrid reactive magnetron sputtering and the surface treatment was performed by spin‐coating CsCO3 solution on the surface of CIGS at room temperature, followed by vacuum annealing at 400°C. The surface chemistry of the CIGS thin film changed after the treatment and the efficiency of respective solar cells improved by more than 30%, mostly driven by an enhancement in open‐circuit voltage. X‐ray photoelectron spectroscopy revealed the depletion of copper and the presence of Cs on the surface of the CIGS thin film. Ultraviolet photoelectron spectroscopy showed the lowering of the valence band maximum by around 0.25 eV after the treatment, which plays a positive role in reducing interfacial recombination. High‐resolution transmission electron microscopy indicates the presence of Cs and depletion of Cu at the grain boundaries of the CIGS thin film. These findings open a low‐cost route for improving the performance of CIGS solar cells by surface modification using a solution process.

Nanomedicine Targeting Cuproplasia in Cancer: Labile Copper Sequestration Using Polydopamine Particles Blocks Tumor Growth In Vivo through Altering Metabolism and Redox Homeostasis.

Copper plays critical roles as a metal active site cofactor and metalloallosteric signal for enzymes involved in cell proliferation and metabolism, making it an attractive target for cancer therapy. In this study, we investigated the efficacy of polydopamine nanoparticles (PDA NPs), classically applied for metal removal from water, as a therapeutic strategy for depleting intracellular labile copper pools in triple-negative breast cancer models through the metal-chelating groups present on the PDA surface. By using the activity-based sensing probe FCP-1, we could track the PDA-induced labile copper depletion while leaving total copper levels unchanged and link it to the selective MDA-MB-231 cell death. Further mechanistic investigations revealed that PDA NPs increased reactive oxygen species (ROS) levels, potentially through the inactivation of superoxide dismutase 1 (SOD1), a copper-dependent antioxidant enzyme. Additionally, PDA NPs were found to interact with the mitochondrial membrane, resulting in an increase in the mitochondrial membrane potential, which may contribute to enhanced ROS production. We employed an in vivo tumor model to validate the therapeutic efficacy of PDA NPs. Remarkably, in the absence of any additional treatment, the presence of PDA NPs alone led to a significant reduction in tumor volume by a factor of 1.66 after 22 days of tumor growth. Our findings highlight the potential of PDA NPs as a promising therapeutic approach for selectively targeting cancer by modulating copper levels and inducing oxidative stress, leading to tumor growth inhibition as shown in these triple-negative breast cancer models.

Cuproptosis: Advances in Stimulus-Responsive Nanomaterials for Cancer Therapy.

Cuproptosis, a recently identified non-apoptotic programmed cell death modality, attracts considerable attention in the realm of cancer therapeutics owing to its unique cellular demise mechanisms. Since its initial report in 2022, strategies inducing or amplifying cuproptosis for cancer treatment emerge. The engineering of nano-systems to elicit cuproptosis effectively circumvents constraints associated with conventional small-molecule pharmaceutical interventions, presenting novel prospects for oncological therapy. Stimulus-responsive nanomaterials, leveraging their distinctive spatiotemporal control attributes, are investigated for their role in modulating the induction or augmentation of cuproptosis. In this comprehensive review, the physiological characteristics of cuproptosis, encompassing facets such as copper overload and depletion, coupled with regulatory factors intrinsic to cuproptosis, are expounded upon. Subsequently, design methodologies for stimulus-responsive induction or enhancement of cuproptosis, employing stimuli such as light, ultrasound, X-ray, and the tumor microenvironment, are systematically delineated. This review encompasses intricacies in nanomaterial design, insights into the therapeutic processes, and the associated advantages. Finally, challenges inherent in stimulus-responsive induction/enhancement of cuproptosis are deliberated upon and prospective insights into the future trajectory of copper-mediated cancer therapy are provided.

The Lanseria palaeosol: Implications for palaeoweathering and terrestrial life at ∼2.6 Ga on the Kaapvaal Craton

Archaean palaeosols are a rare record of surface processes on the early Earth. We report on the discovery of a palaeoweathering profile developed at the contact between Mesoarchaean basement and the Neoarchaean to Palaeoproterozoic Transvaal Supergroup along the northern margin of the Johannesburg Dome, Kaapvaal Craton. Granodiorite ∼3.1 Ga old was subjected to weathering just prior to the deposition of the ∼2.6 Ga Black Reef Formation. The ∼25 m-thick profile is dominated by a palaeosaprolite and organic matter-rich palaeosol remnant at the top that experienced erosion upon marine transgression on the Kaapvaal Craton. The Al concentration of the profile, sericite content and chemical index of alteration (CIA) values progressively increase from the parent granodiorite towards the nonconformity as Fe, Mg, Na and Ca concentrations decrease. The decrease of Co and Zn together with Fe and the retention of U, V and Cr collectively suggest a reducing and acidic environment at the time of weathering. Carbonaceous matter with negative δ13C isotopic values (−27 to −23 ‰V-PDB) in the palaeosol is compatible with the former presence of photoautotrophic microbes. Copper depletion and mobility of P may indicate the presence of organic acids that enhanced chemical weathering of the bedrock. The palaeoweathering profile was subjected to diagenetic-hydrothermal K-metasomatism, regional metamorphism and deformation, which collectively transformed the regolith into a quartz-sericite schist, as it presently occurs. Despite these modifications, the original record of palaeoweathering and microbial colonisation is well preserved. Colonisation of a large swath of the Kaapvaal Craton by terrestrial soil-dwelling microbiota at 2.60 Ga is recorded in abundant soil-derived carbonaceous matter in transgressive marine conglomerates that erosively overlie the palaeosol.

207 Effect of Sodium Sulfate in Water and Dietary Bismuth Subsalicylate on Feed and Water Intake, Ruminal Hydrogen Sulfide Concentration, and Trace Mineral Status of Growing Beef Heifers

Abstract Cattle consuming increased concentrations of sulfur (S) are at an increased risk for depletion of copper (Cu) and or S-induced polioencephalomalacia and there are limited mitigation strategies to alleviate risk for cattle consuming high sulfate water. This study evaluated the effects of feeding growing beef heifers bismuth subsalicylate (BSS; 0.0 vs. 0.4% DM basis) when provided water with a low (LS; 346 ± 13) or high (HS; 4,778 ± 263 mg/L) sulfate concentration on dry matter intake (DMI), water intake, ruminal hydrogen sulfide (H2S) concentration, and trace mineral status. Twenty-four beef heifers (221 ± 41 kg) were stratified based on initial liver copper concentrations collected 13 d before the start of the study, into a completely randomized block design using a high forage diet fed for 98 d. Feed and water intake (weekly), ruminal H2S concentration (d 42 and 91), and liver (pre-study and d 91) and serum trace mineral concentrations (d 1, 28, 56, and 91) were evaluated. Initial liver trace-mineral concentration was used as a covariate in the statistical model. Water intake tended to be reduced with the inclusion of BSS (P = 0.10) but was not affected by water sulfate (P = 0.40). Water sulfate and BSS did not affect DMI (P ≥ 0.89), but total S intake increased (P < 0.01) from LS to HS (17.6 to 51.0 g/d) resulting in diets that contained 0.28 and 0.78% S (DM basis). Heifers consuming HS had 1.58 µg/mL more (P < 0.01) ruminal H2S than LS. The inclusion of BSS reduced (P = 0.04) ruminal H2S concentration by 46%. Regardless of water sulfate concentration, heifers fed BSS had lesser liver Cu (average of 4.08 mg/kg) than heifers not provided BSS, and when not provided BSS, HS had lesser Cu than LS (42.2 vs. 58.3; sulfate × BSS, P = 0.02). Serum concentration of Cu did not differ over time for heifers not provided BSS; whereas heifers provided BSS had less serum Cu on d 91 than d 28 and 55 (BSS × time, P < 0.01). The liver concentration of selenium (Se) was reduced (P < 0.01) with BSS inclusion. The Se concentration in serum was not affected by sulfate, BSS, or time (P ≥ 0.16). Bismuth subsalicylate reduces ruminal H2S concentration, but depletes liver Cu and Se. Moreover, sulfate concentration in water does not appear to affect DMI or water intake but reduces liver Cu concentration.

Copper chelation suppresses epithelial-mesenchymal transition by inhibition of canonical and non-canonical TGF-β signaling pathways in cancer

BackgroundMetastatic cancer cells exploit Epithelial-mesenchymal-transition (EMT) to enhance their migration, invasion, and resistance to treatments. Recent studies highlight that elevated levels of copper are implicated in cancer progression and metastasis. Clinical trials using copper chelators are associated with improved patient survival; however, the molecular mechanisms by which copper depletion inhibits tumor progression and metastasis are poorly understood. This remains a major hurdle to the clinical translation of copper chelators. Here, we propose that copper chelation inhibits metastasis by reducing TGF-β levels and EMT signaling. Given that many drugs targeting TGF-β have failed in clinical trials, partly because of severe side effects arising in patients, we hypothesized that copper chelation therapy might be a less toxic alternative to target the TGF-β/EMT axis.ResultsOur cytokine array and RNA-seq data suggested a link between copper homeostasis, TGF-β and EMT process. To validate this hypothesis, we performed single-cell imaging, protein assays, and in vivo studies. Here, we used the copper chelating agent TEPA to block copper trafficking. Our in vivo study showed a reduction of TGF-β levels and metastasis to the lung in the TNBC mouse model. Mechanistically, TEPA significantly downregulated canonical (TGF-β/SMAD2&3) and non-canonical (TGF-β/PI3K/AKT, TGF-β/RAS/RAF/MEK/ERK, and TGF-β/WNT/β-catenin) TGF-β signaling pathways. Additionally, EMT markers of MMP-9, MMP-14, Vimentin, β-catenin, ZEB1, and p-SMAD2 were downregulated, and EMT transcription factors of SNAI1, ZEB1, and p-SMAD2 accumulated in the cytoplasm after treatment.ConclusionsOur study suggests that copper chelation therapy represents a potentially effective therapeutic approach for targeting TGF-β and inhibiting EMT in a diverse range of cancers.

OECD 414 supplementary prenatal developmental toxicity study of sodium molybdate dihydrate in the rat and benchmark dose evaluation

In a continuing investigation of the potential for reproductive and developmental toxicity of molybdenum (Mo), consequent to the previous published OECD studies [1,2] and as directed by the European Chemicals Agency [3], a supplemental rat GLP-compliant Prenatal Developmental Toxicity (PNDT) study was conducted to investigate higher dose levels of sodium molybdate dihydrate (SMD) in an identical study design (OECD 414)[4] to Murray et al. 2014a [1], at dietary concentrations calculated to provide target Mo levels of 80 and 120 mg/kg bw/day (the maximum-tolerated dose). There was no effect on post-implantation loss, litter size, sex ratio or the incidence of external, visceral or skeletal fetal malformations or variations. Fetal weight was reduced proportionate to maternal dose. Minimal differences observed in the ossification status of some extremities of fetuses from females receiving 120 mg Mo/kg bw/day were confirmed as transient by skeletal examination of PND 21 pups from a further group of females receiving the same dose regime. There was no evidence of copper depletion in serum, placenta or liver. A benchmark dose evaluation using continuous and dichotomous approaches by combining the fetal body weight data from this study and the previous study determined that the BMD05 ranged from 47 to 57 mg Mo/kg bw/day, depending on the modelling approach and the BMDL05 estimates ranged from 37 to 47 mg Mo/kg bw/day. These levels are considered a more statistically robust point of departure for risk assessment for reproductive effects than the established NOAEL of 40 mg Mo/kg bw/day.

Sodium molybdate dihydrate does not exhibit developmental or reproductive toxicity in Sprague-Dawley rats maintained on a marginal copper diet

Groups of 24 weanling female Sprague-Dawley rats were administered molybdenum (Mo) as sodium molybdate dihydrate (SMD) in drinking water at target dose levels of 0, 20, or 40 mg Mo/kg bw/day and fed a semi-purified marginal copper (6.2 ppm Cu) AIN-93 G diet for 8 weeks prior to mating, through cohabitation and pregnancy until Gestation Day 21. The objective was to confirm the reproductive and developmental effects of SMD reported by Fungwe et al. (1990) at estimated doses as low as 1.5 mg Mo/kg bw/day in a similarly designed study with marginal Cu diet (6.3 ppm). There were no test material-related effects at 20 or 40 mg Mo/kg bw/day on mortality, clinical observations, body weight, body weight gain, food consumption, estrous cycling, reproductive performance, maternal macroscopic pathology, ovarian or uterine parameters, litter size, resorptions, fetal sex ratio, fetal weight, or external fetal malformations or variations. Water consumption was increased compared to controls at both dose levels during the pre-mating and gestation periods, with no apparent adverse impact. There was no evidence of copper depletion in serum at any dose level. In conclusion, the no-observed-adverse-effect levels (NOAELs) for systemic, maternal reproductive, and developmental toxicity in this marginal Cu diet study are 40 mg Mo/kg bw/day, consistent with the results of guideline developmental and reproductive toxicity studies of SMD. The results of Fungwe et al. were not replicated, even at higher dose levels of Mo, and their inconsistencies with guideline toxicity studies of Mo are not explained by the marginal dietary Cu level.

Surface Chemistry Dictates the Osteogenic and Antimicrobial Properties of Palladium‐, Platinum‐, and Titanium‐Based Bulk Metallic Glasses

AbstractTitanium alloys are commonly used as biomaterials in musculoskeletal applications, but their long‐term efficacy can be limited by wear and corrosion, stress shielding, and bacterial colonization. As a promising alternative, bulk metallic glasses (BMGs) offer superior strength and corrosion resistance, but the influence of their chemical composition on their bioactivity remains largely unexplored. This study, therefore, aims to examine how the surface chemistry of palladium (Pd)‐, platinum (Pt)‐, and titanium (Ti)‐based BMGs can steer their response to biological systems. The chemical composition of BMGs governs their thermophysical and mechanical properties, with Pd‐based BMGs showing exceptional glass‐forming ability suitable for larger implants, and all BMGs exhibiting a significantly lower Young's modulus than Ti‐6Al‐4 V (Ti64), suggesting a potential to reduce stress shielding. Although BMGs feature copper depletion at the near surface, their surface chemistry remains more stable than that of Ti64 and supports blood biocompatibility. Fibrin network formation is heavily dependent on BMGs’ chemical composition and Ti‐based BMGs support thicker fibrin network formation than Ti64. Furthermore, BMGs outperform Ti64 in promoting mineralization of human bone progenitor cells and demonstrate antimicrobial properties against Staphylococcus aureus in a surface chemistry‐dependent manner, thereby indicating their great potential as biomaterials for musculoskeletal applications.

A Paclitaxel Prodrug with Copper Depletion for Combined Therapy toward Triple-Negative Breast Cancer.

Tuning the content of copper is of great significance for the treatment of cancer and neurodegenerative diseases. Herein, we synthesized a redox-responsive paclitaxel (PTX) prodrug by conjugating PTX with a copper chelator through a disulfide bond. The as-fabricated prodrug (PSPA) showed specific chelation toward copper ions and could assemble with distearoyl phosphoethanolamine-PEG2000 to form stable nanoparticles (PSPA NPs) in aqueous media. Upon being internalized by tumor cells, PSPA NPs could respond to high levels of redox-active species inside cells and efficiently release PTX. The copper chelator could increase oxidative stress- and abnormal metabolism-induced cell death through intracellular copper depletion. The combination of chemotherapy and copper depletion therapy generated an enhanced therapeutic outcome toward triple-negative breast cancer with an ignorable systemic toxicity. Our work may provide insight into the combination of metabolic regulation and chemotherapy for combating malignant tumors.

FRI-349 - Depletion of excess liver copper ameliorates liver damage in a mouse model of chronic cholestatic liver disease

FRI-349 - Depletion of excess liver copper ameliorates liver damage in a mouse model of chronic cholestatic liver disease

Multifaceted Roles of Copper ions in Anticancer Nanomedicine.

The significantly increased copper level in tumor tissues and serum indicates the close association of copper ions with tumor development, making copper ions be attractive targets in the development of novel tumor treatment methods. The advanced nanotechnology developed in the past decades provides great potential for tumor therapy, among which Cu-based nanotherapeutic systems have received greater attention. In this review, the multifaceted roles of copper ions in cancer progression would be summarized and the recent advances in the copper-based nanostructures or nanomedicines for different kinds of tumor therapies including copper depletion therapy, copper-based cytotoxins, copper ions-based chemodynamic therapy and its combination with other treatments and copper ions-induced ferroptosis and cuproptosis activation would be discussed. Furthermore, the perspectives for the further development of copper ions-based nanomedicines for tumor therapy and clinic translation are presented by the authors. This article is protected by copyright. All rights reserved.

Macrophages-Cancer Membrane Encapsulated Metal-Organic Frameworks with Copper-Depleting Moiety for Mitochondria-Targeted Therapeutic.

Mitochondria-targeted therapeutics is an attractive approach against energy-dependent cancer. However, the effective mitochondria organelle therapeutics agents still highly desirable. Herein, we report a mitochondria-targeted therapeutics platform, termed CDM@MUiO-DP@MCHM, consisted of macrophages-cancer hybrid membrane (MCHM) encapsulated MUiO-66 metal-organic frameworks (MOFs), which was loaded with microRNA (miRNA) biomarker detection probe (DP) for cancer diagnosis and copper-depleting moiety (CDM) for mitochondrial copper depletion to suppress cancer growth. Using nude mice bearing MCF-7 as model, after injected intravenously via the caudal vein of mice, the encapsulation of MCHM can not only greatly enhance the cancer homing-targeting ability of the nanoparticles (NPs), but also endows the NPs the immune escape capacity to extend the circulation time. The miRNA-21 biomarker could be detected by the fluorescence signal for diagnosis, while the CDM induced energy deficiency of and compromised mitochondria membrane potential, leading to apoptosis of the cancer cell. The good performance of CDM@MUiO-DP@MCHM suggest the great potential mitochondria organelle therapeutics. This article is protected by copyright. All rights reserved.

ARBM101 (Methanobactin SB2) Drains Excess Liver Copper via Biliary Excretion in Wilson’s Disease Rats

Excess copper causes hepatocyte death in hereditary Wilson's disease (WD). Current WD treatments by copper-binding chelators may gradually reduce copper overload; they fail, however, to bring hepatic copper close to normal physiological levels. Consequently, lifelong daily dose regimens are required to hinder disease progression. This may result in severe issues due to nonadherence or unwanted adverse drug reactions and also due to drug switching and ultimate treatment failures. This study comparatively tested bacteria-derived copper binding agents-methanobactins (MBs)-for efficient liver copper depletion in WD rats as well as their safety and effect duration. Copper chelators were tested invitro and invivo in WD rats. Metabolic cage housing allowed the accurate assessment of animal copper balances and long-term experiments related to the determination of minimal treatment phases. We found that copper-binding ARBM101 (previously known as MB-SB2) depletes WD rat liver copper dose dependently via fecal excretion down to normal physiological levels within 8 days, superseding the need for continuous treatment. Consequently, we developed a new treatment consisting of repetitive cycles, each of ∼1 week of ARBM101 applications, followed by months of in-between treatment pauses to ensure a healthy long-term survival in WD rats. ARBM101 safely and efficiently depletes excess liver copper from WD rats, thus allowing for short treatment periods as well as prolonged in-between rest periods.

Zinc-Carnosine Metallodrug Network as Dual Metabolism Inhibitor Overcoming Metabolic Reprogramming for Efficient Cancer Therapy.

The targeting of tumor metabolism as a novel strategy for cancer therapy has attracted tremendous attention. Herein, we develop a dual metabolism inhibitor, Zn-carnosine metallodrug network nanoparticles (Zn-Car MNs), which exhibits good Cu-depletion and Cu-responsive drug release, causing potent inhibition of both OXPHOS and glycolysis. Notably, Zn-Car MNs can decrease the activity of cytochrome c oxidase and the content of NAD+, so as to reduce ATP production in cancer cells. Thereby, energy deprivation, together with the depolarized mitochondrial membrane potential and increased oxidative stress, results in apoptosis of cancer cells. In result, Zn-Car MNs exerted more efficient metabolism-targeted therapy than the classic copper chelator, tetrathiomolybdate (TM), in both breast cancer (sensitive to copper depletion) and colon cancer (less sensitive to copper depletion) models. The efficacy and therapy of Zn-Car MNs suggest the possibility to overcome the drug resistance caused by metabolic reprogramming in tumors and has potential clinical relevance.

Tetrathiomolybdate Decreases the Expression of Alkaline Phosphatase in Dermal Papilla Cells by Increasing Mitochondrial ROS Production

Dermal papilla cells (DPCs) play important roles in hair growth regulation. However, strategies to regrow hair are lacking. Here, global proteomic profiling identified the tetrathiomolybdate (TM)-mediated inactivation of copper (Cu) depletion-dependent mitochondrial cytochrome c oxidase (COX) as the primary metabolic defect in DPCs, leading to decreased Adenosine Triphosphate (ATP) production, mitochondrial membrane potential depolarization, increased total cellular reactive oxygen species (ROS) levels, and reduced expression of the key marker of hair growth in DPCs. By using several known mitochondrial inhibitors, we found that excessive ROS production was responsible for the impairment of DPC function. We therefore subsequently showed that two ROS scavengers, N-acetyl cysteine (NAC) and ascorbic acid (AA), partially prevented the TM- and ROS-mediated inhibition of alkaline phosphatase (ALP). Overall, these findings established a direct link between Cu and the key marker of DPCs, whereby copper depletion strongly impaired the key marker of hair growth in the DPCs by increasing excessive ROS production.

Proteomics revealed the crosstalk between copper stress and cuproptosis, and explored the feasibility of curcumin as anticancer copper ionophore.

As an essential micronutrient element in organisms, copper controls a host of fundamental cellular functions. Recently, copper-dependent cell growth and proliferation have been defined as "cuproplasia". Conversely, "cuproptosis" represents copper-dependent cell death, in a nonapoptotic manner. So far, a series of copper ionophores have been developed to kill cancer cells. However, the biological response mechanism of copper uptake has not been systematically analyzed. Based on quantitative proteomics, we revealed the crosstalk between copper stress and cuproptosis in cancer cells, and also explored the feasibility of curcumin as anticancer copper ionophore. Copper stress not only couples with cuproptosis, but also leads to reactive oxygen species (ROS) stress, oxidative damage and cell cycle arrest. In cancer cells, a feedback cytoprotection mechanism involving cuproptosis mediators was discovered. During copper treatment, the activation of glutamine transporters and the loss of Fe-S cluster proteins are the facilitators and results of cuproptosis, respectively. Through copper depletion, glutathione (GSH) blocks the cuproptosis process, rescues the activation of glutamine transporters, and prevents the loss of Fe-S cluster proteins, except for protecting cancer cells from apoptosis, protein degradation and oxidative damage. In addition, the copper ionophore curcumin can control the metabolisms of lipids, RNA, NADH and NADPH in colorectal cancer cells, and also up-regulates positive cuproptosis mediators. This work not only established the crosstalk between copper stress and cuproptosis, but also discolored the suppression and acceleration of cuproptosis by GSH and curcumin, respectively. Our results are significant for understanding cuproptosis process and developing novel anticancer reagents based on cuproptosis.

Copper Depletion Strongly Enhances Ferroptosis via Mitochondrial Perturbation and Reduction in Antioxidative Mechanisms.

Copper serves as a co-factor for a host of metalloenzymes, particularly cytochrome c oxidase (COX). Although it is known that impaired COX function can lead to the excessive accumulation of reactive oxygen species (ROS), the mechanisms underlying how copper depletion leads to cell damage are poorly understood. Here, we have investigated the role of copper depletion during ferroptosis. The bathocuproinedisulfonic (BCS) treatment depolarized the mitochondrial membrane potential, increased the total cellular ROS levels, stimulated oxidative stress, and reduced the glutathione levels. Moreover, the depletion of copper limited the protein expression of glutathione peroxidase 4 (GPX4), which is the only enzyme that is known to prevent lipid peroxidation. Furthermore, we found that copper depletion decreased the sensitivity of the dermal papilla cells (DPCs) to erastin (an inducer of ferroptosis), and the ferroptosis inhibitor ferrostatin-1 (Fer-1) partially prevented BCS-mediated cell death. Overall, these findings establish a direct link between copper and ferroptosis; BCS-mediated copper depletion strongly enhances ferroptosis via mitochondrial perturbation and a reduction in antioxidative mechanisms.