Elevated Iron Levels Research Articles

Cellular iron depletion enhances behavioral rhythm by limiting brain Per1 expression in mice.

Disturbances in the circadian rhythm are positively correlated with the processes of aging and related neurodegenerative diseases, which are also associated with brain iron accumulation. However, the role of brain iron in regulating the biological rhythm is poorly understood. In this study, we investigated the impact of brain iron levels on the spontaneous locomotor activity of mice with altered brain iron levels and further explored the potential mechanisms governing these effects invitro. Our results revealed that conditional knockout of ferroportin 1 (Fpn1) in cerebral microvascular endothelial cells led to brain iron deficiency, subsequently resulting in enhanced locomotor activity and increased expression of clock genes, including the circadian locomotor output cycles kaput protein (Clock) and brain and muscle ARNT-like 1 (Bmal1). Concomitantly, the levels of period circadian regulator 1 (PER1), which functions as a transcriptional repressor in regulating biological rhythm, were decreased. Conversely, the elevated brain iron levels in APP/PS1 mice inhibited autonomous rhythmic activity. Additionally, our findings demonstrate a significant decrease in serum melatonin levels in Fpn1cdh5 -CKO mice compared with the Fpn1flox/flox group. In contrast, APP/PS1 mice with brain iron deposition exhibited higher serum melatonin levels than the WT group. Furthermore, in the human glioma cell line, U251, we observed reduced PER1 expression upon iron limitation by deferoxamine (DFO; iron chelator) or endogenous overexpression of FPN1. When U251 cells were made iron-replete by supplementation with ferric ammonium citrate (FAC) or increased iron import through transferrin receptor 1 (TfR1) overexpression, PER1 protein levels were increased. Additionally, we obtained similar results to U251 cells in mouse cerebellar astrocytes (MA-c), where we collected cells at different time points to investigate the rhythmic expression of core clock genes and the impact of DFO or FAC treatment on PER1 protein levels. These findings collectively suggest that altered iron levels influence the circadian rhythm by regulating PER1 expression and thereby modulating the molecular circadian clock. In conclusion, our study identifies the regulation of brain iron levels as a potential new target for treating age-related disruptions in the circadian rhythm.

Effect of augmented nutrient composition and fertigation system on biomass yield and cannabinoid content of medicinal cannabis (Cannabis sativa L.) cultivation.

Growing evidence underscores the role of nutrients and fertigation systems in soilless production, influencing medicinal cannabis biomass and secondary metabolite content. This study delves into the impact of enhanced nutrient regimes on the 'ionome' and its ramifications for biomass and cannabinoid production in medicinal cannabis, comparing two distinct fertigation systems: recirculation and drain-to-waste. Notably, we assess the optimal harvest time for maximizing profitability. In comparing the experimental variant with elevated levels of phosphorus (P), potassium (K), and iron (Fe) in the nutrient solution to the control variant, we observe distinct patterns in element composition across stems, leaves, and flowers, with significant differences between fertigation systems. Total nitrogen content was determined through the Kjeldahl method. Flame atomic absorption spectrometry (FAAS) and inductively coupled plasma optical emission spectrometry (ICP-OES) were employed for elemental analysis. Cannabinoid identification and quantification used high-performance liquid chromatography with a diode-array detector (HPLC/DAD). Followed statistical analyses included ANOVA and Tukey's HSD test. Although the augmented nutrient regimen does not substantially increase plant biomass, interesting differences emerge between the two fertigation systems. The recirculation fertigation system proves more profitable during the recommended harvest period. Nonetheless, the altered nutrient regime does not yield statistically significant differences in final inflorescence harvest mass or cannabinoid concentrations in medicinal cannabis. The choice of fertigation system influences the quantity and quality of harvested inflorescence. To optimize the balance between the dry biomass yield of flowers and cannabinoid concentration, primarily total THC yield (sum of tetrahydrocannabinolic acid, Δ9-tetrahydrocannabinol, and Δ8-tetrahydrocannabinol), we propose the 11th week of cultivation as the suitable harvest time for the recirculation system. Importantly, the recirculation system consistently outperformed the drain-to-waste system, especially after the ninth week, resulting in significantly higher total THC yields. Enriched nutrition, when compared with control, increased THC yield up to 50.7%, with a remarkable 182% surge in the recirculation system when compared with the drain-to-waste system.

Assessing groundwater quality, health risks, and policy implications: A case study of West Medinipur District, West Bengal, India

AbstractGroundwater is a vital freshwater resource for various uses, including drinking, irrigation, and industry needs. The increasing dependence of India on groundwater due to dwindling surface water sources and climate extremes has led to concerns about its availability and quality. This study focuses on the groundwater quality and associated human health risks in the West Medinipur district of West Bengal, India. The region, situated between the Kangsabati and Subarnarekha rivers, faces challenges from anthropogenic activities such as agricultural practices and industrial pollution. A comprehensive analysis of physicochemical parameters was conducted on well water samples collected during the monsoon and post‐monsoon periods. The study examined the suitability of groundwater for drinking and irrigation purposes. Water quality indices were calculated to assess its fitness for human consumption. Despite several parameters meeting the water quality standards, elevated iron (Fe) and ammonia (NH4+) levels were observed, potentially impacting groundwater use for drinking. Similarly, groundwater's suitability for irrigation was evaluated based on parameters like sodium adsorption ratio (SAR) and sodium percentage (Na%). The groundwater types dominated by magnesium bicarbonate and the quality could affect soil structure and crop productivity in certain areas. The elevated levels of Fe and NH4+ showed potential human health risks, indicating the need to mitigate adverse effects on human well‐being through appropriate policy and other interventions. The study highlights the complexity of groundwater quality in the study area. It underscores the necessity for comprehensive water policy and management strategies to safeguard water resources and human health in rural India. The findings emphasize the importance of ongoing monitoring and intervention measures to ensure the sustainability and safety of groundwater for present and future generations.

Methyltransferase-like 3 (METTL3) Epigenetically Modulates Glutathione Peroxidase 4 (GPX4) Expression to Affect Asthma.

Asthma, a prevalent chronic airway inflammatory condition, poses a significant health challenge. In this study, we delved into the regulatory mechanisms governing asthma, focusing on Methyltransferase-like 3 (METTL3). Through an ovalbumin (OVA)-induced mouse model and interleukin-13 (IL-13)-induced cell model, we mimicked the in vivo and in vitro functions of METTL3 in asthma. Our research revealed that METTL3 expression significantly decreased in asthma-induced mice and IL-13-stimulated cells compared to the control group. Moreover, METTL3 overexpression enhanced bronchial epithelial cell viability and proliferation. Mechanistically, we observed elevated levels of total iron, Fe2+, malondialdehyde (MDA), lipid reactive oxygen species (ROS), alongside reduced glutathione (GSH) levels in IL-13-stimulated cells. Remarkably, METTL3 overexpression counteracted these effects, suggesting a pivotal role in mitigating asthma-related oxidative stress. Furthermore, our study highlighted the involvement of N6-methyladenosine methylation (m6A) modification, where METTL3 regulated the m6A modification of glutathione peroxidase 4 (GPX4) RNA, impacting RNA stability. Knockdown of METTL3 suppressed m6A modification on GPX4 RNA, impairing its stability and contributing to IL-13-induced ferroptosis. Interestingly, METTL3 overexpression not only inhibited cell ferroptosis but also alleviated asthma symptoms. Our findings shed light on the epigenetic regulation of asthma through METTL3-mediated m6A modification, offering potential therapeutic avenues for this prevalent inflammatory disease.

Curcumin-polydopamine nanoparticles alleviate ferroptosis by iron chelation and inhibition of oxidative stress damage.

Ferroptosis, characterized by elevated iron levels and lipid peroxidation (LPO), is a recently identified regulatory mechanism of cell death. Its substantial involvement in ischemic tissue injury, neurodegenerative disorders, and cancer positions ferroptosis inhibition as a promising strategy for managing these diverse diseases. In this study, we introduce curcumin-polydopamine nanoparticles (Cur-PDA NPs) as an innovative ferroptosis inhibitor. Cur-PDA NPs demonstrate remarkable efficacy in chelating both Fe2+ and Fe3+in vitro along with scavenging free radicals. Cur-PDA NPs were found to efficiently mitigate reactive oxygen species, reduce Fe2+ accumulation, suppress LPO, and rejuvenate mitochondrial function in PC12 cells. Thus, these NPs can act as potent therapeutic agents against ferroptosis, primarily via iron chelation and reduction of oxidative stress.

Сочетание синдрома Жильбера и гемохроматоза у пациентки с цитолизом и гипербилирубинемией

Aim. To present a clinical case of the combination of Gilbert's syndrome and primary hemochromatosis. Key points. The article presents a clinical example demonstrating the complexity of diagnosing diseases caused by genetic polymorphism. Examination of the patient revealed impaired liver function, elevated levels of hemoglobin, serum iron, and transferrin saturation with iron, which led to suspicion of Gilbert's syndrome and primary hemochromatosis. These diagnoses were confirmed by genetic testing. Conclusion. Diagnosis of the causes of liver damage requires first and foremost exclusion of the most likely ones, but it is important to keep in mind the possibility of continuing the search for the causes of the disease using specific diagnostic methods. Keywords: Gilbert's syndrome, hereditary hemochromatosis, cytolysis, hyperbilirubinemia, hepatitis.

Anthocyanins-rich cranberry extract attenuates DSS-induced IBD in an intestinal flora independent manner

Cranberry is abundantly rich in anthocyanins, a type of flavonoid with potent antioxidant properties and the resistance against certain diseases. In this study, anthocyanin-rich cranberry extract was extracted, purified, and its components were analyzed. 92.18 % of anthocyanins was obtained and the total content of anthocyanins was 302.62 mg/g after AB-8 resin purification. Quantification analysis showed that the extract mainly contained cyanidin-3-galactoside, procyanidin B2 and procyanidin B4. Then we explored its effects on dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) in mice. The supplementation of cranberry extract resulted in an alleviation of IBD symptoms, evidenced by improvements in the disease activity index (DAI), restoration of colon length and colonic morphology. Cranberry extract reversed the elevated iron and malondialdehyde (MDA) levels and restored glutathione (GSH) levels in IBD mice. Further analysis revealed that cranberry modulated ferroptosis-associated genes and reduced expression of pro-inflammatory cytokines. Although cranberry influenced the intestinal flora balance by reducing Proteobacteria and Escherichia-Shigella, and increasing Lactobacillus, as well as enhancing SCFAs content, these effects were not entirely dependent on intestinal flora modulation, as indicated by antibiotic intervention and fecal microbiota transplantation (FMT) experiments. In conclusion, our findings suggest that the beneficial impact of cranberry extract on IBD may primarily involve the regulation of colonic ferroptosis, independent of significant alterations in intestinal flora.

Ferroptosis: a new perspective on the pathogenesis of radiation-induced cataracts.

Ionizing radiation is a significant risk factor for cataracts, but the pathogenesis of radiation-induced cataracts remains incompletely understood. Ferroptosis, an iron-dependent form of programmed cell death discovered in recent years, has gained increasing attention for its role in various diseases. This article systematically reviews research progress on ionizing radiation, ferroptosis, age-related cataracts, and radiation-induced cataracts. It proposes the "ferroptosis hypothesis" for the pathogenesis of radiation-induced cataracts. Through ionization and oxidative stress effects, ionizing radiation leads to elevated free iron levels and exacerbated lipid peroxidation in lens cells, activating the ferroptosis pathway and resulting in lens opacity. The involvement of ferroptosis in the development of age-related cataracts suggests that it may also be an important pathogenic mechanism of radiation-induced cataracts. Targeting the ferroptosis pathway may be a novel strategy for preventing and treating radiation-induced cataracts. Furthermore, developing new ferroptosis-specific inhibitors with improved targeting and pharmacokinetic properties is also an essential direction for research on preventing and treating radiation-induced cataracts. The study of ferroptosis provides new insights into the mechanism and management of radiation-induced cataracts, potentially transforming radiation-induced cataracts from "inevitable" to "preventable and treatable."

Synergy between isobavachalcone and doxorubicin suppressed the progression of anaplastic thyroid cancer through ferroptosis activation.

The objective of this study was to explore the effects and mechanisms of the combination of isobavachalcone (IBC) and doxorubicin (DOX) on the progression of anaplastic thyroid cancer (ATC). Cell viability of 8505C and CAL62 cells was observed by CCK-8 assay. Kits were used to detect the presence of reactive oxygen species (ROS), glutathione (GSH), malondialdehyde (MDA), and cellular iron. Protein expression of solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) was detected using western blot, and CD31 was detected through immunofluorescence. Tumor xenograft models of 8505C cells were constructed to observe the effect of IBC and DOX on ATC growth in vivo. The co-administration of IBC and DOX exhibited a synergistic effect of suppressing the growth of 8505C and CAL62 cells. The concurrent use of IBC and DOX resulted in elevated iron, ROS, and MDA levels, while reducing GSH levels and protein expression of SLC7A11 and GPX4. However, the Fer-1 ferroptosis inhibitor effectively counteracted this effect. In vitro and in vivo, the inhibitory effect on ATC cell proliferation and tumor growth was significantly enhanced by the combination of IBC and DOX. The combination of IBC and DOX can inhibit the growth of ATC by activating ferroptosis, and might prove to be a potent chemotherapy protocol for addressing ATC.

Electroacupuncture improves cognitive function in APP/PS1 mice by inhibiting oxidative stress related hippocampal neuronal ferroptosis

BackgroundElectroacupuncture, recognized as a crucial non-pharmacological therapeutic approach, has demonstrated notable efficacy in enhancing cognitive function among Alzheimer's disease (AD) patients. This study aimed to investigate the neuroprotective properties of electroacupuncture in APP/PS1 mice with AD. MethodsA total of thirty APP/PS1 mice were randomly assigned to three groups: the Alzheimer's disease group (AD), the electroacupuncture treatment group (EA), and the ferroptosis inhibitor deferasirox treatment group (DFX). Additionally, ten C57BL/6 mice were included as a control group (Control). In the EA group, mice underwent flat needling at Baihui and Yintang, as well as point needling at Renzhong, once daily for 15 min each time. In the DFX group, mice received intraperitoneal injections of deferasirox at a dosage of 100 mg/kg/day. Following the 28-day treatment period, behavioral evaluation, morphological observation of neurons, and detection of neuronal ferroptosis were conducted. ResultsThe electroacupuncture treatment demonstrated a significant improvement in spatial learning, memory ability, and neuronal damage in mice with AD. Analysis of neuronal ferroptosis markers indicated that electroacupuncture interventions reduced the elevated levels of malondialdehyde, iron, and ptgs2 expression, while also increasing superoxide dismutase activity, Ferroportin 1 and glutathione peroxidase 4 expression. Moreover, the regulatory impact of electroacupuncture on ferroptosis may be attributed to its ability to enhance the expression and nuclear translocation of Nrf2. ConclusionsThis study suggested that electroacupuncture could inhibit the neuronal ferroptosis by activating the antioxidant function in neurons through p62/Keap1/Nrf2 signal pathway, thereby improve the cognitive function of AD mice by the neuronal protection effect.

Pseudolaric acid B triggers ferritinophagy and ferroptosis via upregulating NCOA4 in lung adenocarcinoma cells.

Ferroptosis is a novel subtype of programmed cell death caused by iron-dependent lipid peroxidation and excessive reactive oxygen species (ROS) production. Small-molecule ferroptotic drugs have the probability of selectively targeting the specific features of aggressive tumor cells. In particular, pseudolaric acid B (PAB) triggered ferroptosisin breast cancer cells. The aim of this study is to explore the antitumor effect of PAB on A549 cells and provide a theoretical basis for the further development and clinical application of PAB. First, relevant databases were used to predict of target genes related to PAB, Then, EdU proliferation assay, colony formation and wound-healing assays were applied to calculate A549 cells proliferative abilities. Measurement of ferrous iron, lipid peroxidation, ROS, malondialdehyde (MDA) and glutathione (GSH) were utilized to explore the relevant mechanism. We showed that PAB decreased the viability of lung adenocarcinoma cells in vitro, which was accompanied by abnormally elevated levels of intracellular ferrous iron and overproduction of lipid reactive oxidate species (L-ROS). In turn, deferoxamine (DFO) significantly rescued PAB-induced lipid peroxidation. PAB also improved the intracellular labile iron pool by promoting ferritin autophagy via the upregulation of the nuclear receptor coactivator 4 (NCOA4). Moreover, silencing of NCOA4 alleviated PAB-inducedferroptotic death and reduced the levels of intracellular ferrous iron. In summary, PAB-triggered ferroptosis in lung adenocarcinoma cells by enhancing ferritinophagy. thus, PAB is a potential therapeutic agent for lung adenocarcinoma.

Serum Iron, Ferritin, Erythroferrone and their Inter-Correlation In Adult Cigarette Smokers: A Case-Control study

Cigarette smoking is one of the leading causes of lung and blood cancers worldwide. A higher risk of pulmonary injury is associated with elevated lung iron and ferritin levels. Erythroferrone (ERFE) is a newly discovered tumor necrosis factor (TNF) superfamily member produced by erythroblasts during erythropoiesis. Eventually, it suppresses hepcidin during enhanced erythropoietic activity. The current study investigated the relationship between serum ferritin, iron, and erythroferrone concentrations among adult smokers. This study included 76 selected adult cigarette smokers and 48 healthy individuals (non-smokers) as a control group. The enzyme-linked immunosorbent assay (ELISA) and chemiluminescent immunoassay (CLIA) biochemical techniques were used to accurately measure serum iron, ferritin, and erythroferrone. Cigarette smokers (CS) had a mean age of 42.75±2.92 years, while healthy control (HC) subjects had a mean age of 37.36±2.7 years. The findings have shown a significant increase in serum iron, ferritin, and EREF levels in CS (119± 4.30, 131.1±6.58, and 75.02±2.09) compared with HC (98.67±7.84, 85.50±4.12, and 65.74±2.25) respectively. This study has shown a significant positive correlation between serum EREF (P=0.0072, r=0.4089) and the duration of smoking <20 years and a moderate association between serum iron (P=0.1907, r=0.4511) and the duration of smoking <40 years. For the first time, this outcome shows increased serum EREF and overexpression by erythroblasts in subjects associated with smoking habits, along with increased serum iron and ferritin levels. An indicative correlation is found between EREF overexpression, iron overload, and hyperferritinemia with reference to the duration of smoking. The study also illustrates the interconnection between iron metabolism perturbations and ERFE overexpression induced by cigarette smoking.

Enhanced understanding of the involvement of ferroptosis in tumorigenesis: A review of recent research advancements

<p class="MsoNormal" style="margin-top: 12pt; text-align: justify;"><span lang="EN-US" style="font-family: arial, helvetica, sans-serif;">Ferroptosis, a recently identified form of programmed cell death, is characterized by the accumulation of lipid peroxidation, reactive oxygen species, and elevated free iron levels, involving the regulation of glutathione metabolism, iron metabolism, lipid metabolism, and oxidative stress biology. Tumor metastasis, a critical hallmark of malignancy and a key contributor to cancer recurrence and mortality, has been extensively linked to iron dysregulation, highlighting the potential of agents inducing iron-mediated cell death as promising strategies for preventing and treating metastasis. This review offers a comprehensive understanding the regulatory mechanisms underlying ferroptosis and its crucial role in the three distinct stages of metastasis: invasion, circulation, and colonization.</span></p>

Enhanced understanding of the involvement of ferroptosis in tumorigenesis: A review of recent research advancements

<p class="MsoNormal" style="margin-top: 12pt; text-align: justify;"><span lang="EN-US" style="font-family: arial, helvetica, sans-serif;">Ferroptosis, a recently identified form of programmed cell death, is characterized by the accumulation of lipid peroxidation, reactive oxygen species, and elevated free iron levels, involving the regulation of glutathione metabolism, iron metabolism, lipid metabolism, and oxidative stress biology. Tumor metastasis, a critical hallmark of malignancy and a key contributor to cancer recurrence and mortality, has been extensively linked to iron dysregulation, highlighting the potential of agents inducing iron-mediated cell death as promising strategies for preventing and treating metastasis. This review offers a comprehensive understanding the regulatory mechanisms underlying ferroptosis and its crucial role in the three distinct stages of metastasis: invasion, circulation, and colonization.</span></p>

Downregulation of SLC7A11 in MDS-SF3B1 Patients Induces a High Sensitivity of Erythroid Precursors to Ferroptosis

Introduction Myelodysplastic syndromes with SF3B1 mutation (MDS- SF3B1) represent a subtype of hematopoietic stem cell disorders characterized by bone marrow erythroid dysplasia and ring sideroblasts. They are associated with patient-impacting anemia and iron overload. Adequate iron levels are crucial for heme synthesis during erythropoiesis. An elevated iron level can enhance cell sensitivity to ferroptosis, a non-apoptotic iron-dependent form of cell death caused by lipid peroxidation. Recent studies have demonstrated that normal erythroblasts (EBs) are resistant to ferroptosis, expressing highly Glutathione Peroxidase 4 (GPX4), a ferroptosis suppressor, which requires Glutathione (GSH) as a cofactor. We hypothesize here that a loss of resistance to ferroptosis could occur in MDS- SF3B1 EBs, contributing to ineffective erythropoiesis Methods and results We first used UT7-EPO and HUDEP-2 as cell models of in vitro erythropoiesis to study the effects of SF3B1 knockdown (KD) using a shRNA lentiviral-based strategy. In both cell lines, we observed after SF3B1 KD a higher level of ferroptosis, as assessed using BODIPY 581/591 C11 staining, both at basal state and after exposure to Ras-Selective Lethal 3 (RSL3), which degrades GPX4 and L-Buthionine- (S, R)-sulfoximine (BSO), which inhibits the cysteine import and subsequently GSH synthesis. In EBs derived from CD34 + cells, SF3B1 KD induced a delayed differentiation together with an increased level of ferroptosis, and a decreased level of GSH. Since GSH production requires import of Cystine through the Solute Carrier family 7 member 11 ( SLC7A11) transporters, we quantified this transporter by Western Blot and found it decreased in comparison to control. The use of γ-Glutamyl-Cysteine, which bypasses SLC7A11 for GSH synthesis, decreased ferroptosis both at basal state and after RSL3 or BSO exposure.In normal EBs, GPX4 inhibition is not enough to induce a high ferroptosis. We hypothesized that, after SF3B1 KD, the decreased GSH lowered the “trigger” for ferroptosis, but that an additional stimulus was required. Free iron appeared to be a good candidate since (i) it plays the central role in ferroptosis, (ii) an excess of free iron is observed in MDS- SF3B1 EBs. We exposed EBs after SF3B1 KD to free iron. We observed increased intracytoplasmic reactive oxygen species level (ROS), measured using the H2DCFDA probe and increased membrane lipid peroxidation as well as accumulation of intracytoplasmic iron assessed by FerroOrange probe while it decreased rapidly after a peak in the control conditions. Quantifying ferroportin revealed a decreased expression after SF3B1 KD. α-Tocopherol, that reverts lipid peroxidation and N-Acetyl-Cysteine, that decreased intracellular ROS, restored ferroportin expression, iron excretion andoxidative stress. At last, we used primary bone marrow cells from 3 MDS- SF3B1 patients and confirmed the phenotype observed in other cell models characterized by (i) an excess of basal ferroptosis, (ii) a decreased SLC7A11 and ferroportin expression and (iii) an accumulation of intracellular free iron in comparison with normal EBs. Conclusion In summary, we show that SF3B1 KD or mutation in primary cells from patients, impaired the natural resistance of EBs to ferroptosis together with a defect in erythroid differentiation and an increased mortality, potentially contributing to anemia. The heightened susceptibility to ferroptosis is associated with downregulation of SLC 7A11 expression, leading to a decreased GSH synthesis. Moreover, the high level of ROS is associated with a decreased ferroportin expression, leading to accumulation of intracellular free iron and providing a permanent “fuel” for membrane lipid peroxidation that could be reverted by ferroptosis-blocking agents such as Vitamin E or Deferasirox. Both showed promise in restoring erythroid differentiation, preventing EB cell death, and may serve as an interesting therapeutic strategy for treating MDS- SF3B1 patients.

Ineffective Erythropoiesis Negatively Impacts the Erythroblastic Island in Sickle Cell Disease

Sickle cell disease (SCD) is a genetic recessive disorder, characterized by painful episodes of vaso-occlusion, chronic hemolytic anemia, and progressive organ failure. It is one of the most common and severe monogenic diseases worldwide. SCD is caused by a point mutation in the β-globin gene, leading to the expression of an abnormal hemoglobin (HbS) that polymerizes under hypoxic conditions, driving red blood cells (RBC) sickling. Peripheral RBCs are extensively studied in SCD, as hemolysis is the major contributor to anemia, but little is known about erythroid differentiation in this pathology. We recently demonstrated the occurrence of ineffective erythropoiesis in the bone marrow (BM) of SCD patients, characterized by the death of a significant proportion of erythroblasts between the polychromatic and the orthochromatic stages. We hypothesize that this increased cell death could negatively impact the central macrophages of the erythroblastic islands (EBIs) as they would engulf high numbers of hemoglobin-rich apoptotic erythroblasts and undergo ferroptosis because of excess iron. To investigate our hypothesis, we used an in vitro system whereby CD14 + monocytes were isolated from peripheral blood and cultured in media supplemented with M-CSF and dexamethasone. This led them to differentiate towards anti-inflammatory tissue macrophages, with an “EBI-like” phenotype, expressing markers such as CD16, CD163, CD169, CD206. Phagocytosis of high numbers of RBCs by “EBI-like” macrophages led to increased cell death, reactive oxygen species, and lipid peroxidation, and to the upregulation of genes involved in ferroptosis pathways such as HO-1 and SLC7A11. This was circumvented by the ferroptosis inhibitors ferrostatin-1 and liproxtatin-1. Furthermore, phagocytosis of apoptotic in vitro-generated SCD erythroblasts also induced ferroptosis in EBI-like macrophages, with increased cell death and lipid peroxidation in a time-dependent manner. Interestingly, while this was the case with hemoglobin-rich, terminally differentiating erythroblasts, phagocytosis of cells without hemoglobin, such as K562 or GPA - progenitor cells did not affect cell viability, indicating that the observed ferroptosis was due to the iron content of the internalized cells. We investigated the effect of increased RBC phagocytosis on the phenotype of surviving macrophages and found an M2 to M1 shift indicating a switch to a pro-inflammatory phenotype under the condition of elevated levels of intracellular iron and oxidative stress. We challenged these findings in vivo using the Townes humanized SCD mouse model. Data on 4-month-old mice showed the expected switch in erythroid production from steady-state erythropoiesis in the BM to a stress erythropoiesis in the spleen. This was associated with a decrease in the number of EBI macrophages (F4/80 HighVCAM+CD169+Ly6G-) in the BM of SS mice when compared to AS and AA mice, and an expansion of red pulp macrophages (RPMs) F4/80 HighCD169+VCAM+. Using a flow cytometry-based assay we found high lipid peroxidation levels in BM EBI macrophages of SS mice as compared to AA, suggesting occurrence of ferroptosis in these cells. Our study suggests that IE in SCD may contribute to anemia, directly by cell death of differentiating erythroblasts, and indirectly by disrupting the erythroblastic island or by inducing its collapse following ferroptosis of the central macrophage. Our work with the Townes mouse model and BM samples from SCD patients will help to elucidate this hypothesis.

Myelodysplastic Syndromes Benefit from Iron-Restricted Bone Marrow Transplant in Preclinical Mouse Model

Background: Hematopoietic stem cell transplantation (HSCT) remains the only fully curative therapy for diseases such as b-thalassemia, sickle cell disease, MDS and AML and represents a relevant procedure for gene therapy .Tissue iron overload and non-transferrin bound iron (NTBI) have emerged as adverse prognostic factors for post-transplant survival. Recently, we showed that - in line with observations in transplanted individuals - preparatory chemotherapy conditioning with a combination of busulfan and fludarabine in wild-type mice resulted in elevated serum iron and transferrin (Tf) saturation and elicited NTBI formation. Tf saturation and NTBI levels increased proportionally with conditioning intensity. Interestingly, we found that pre-conditioning TMPRSS6 silencing through a GalNAc-conjugated siRNA - by increasing hepcidin levels - reduced the peri-transplant elevation of Tf saturation and limited the appearance of NTBI. These results proved that NTBI formation is induced by conditioning chemotherapy in the peri-trasplant period independently of the presence of pre-existing iron overload. Aims & Methods: Here we asked whether these observations hold true in conditions hallmarked by pre-existing iron overload and elevated NTBI and how iron restriction impacts on bone marrow transplant (BMT) outcome. To this aim, we applied the GalNAc TMPRSS6 siRNA as iron restriction strategy in a mouse model of myelodysplastic syndromes (MDS) and assessed its ability to reduce conditioning-elicited NTBI formation as well as its potential benefit on BMT. After TMPRSS6 siRNA administration, CD45.2 + MDS mice received conditioning chemotherapy and transplant of CD45.1 + wild-type bone marrow cells. Systemic iron parameters, engraftment and chimerism were then analyzed. Results: Importantly, conditioning exacerbated the elevated systemic iron levels in MDS mice, which was prevented by pre-conditioning TMPRSS6 silencing. Similar to our observation in wild-type mice, recipient CD45.2 + MDS mice showed a superior blood chimerism, resulting from increased bone marrow and spleen engraftment of CD45.1 + donor cells. Multilineage engraftment in bone marrow, spleen and peripheral blood was superior in iron-restricted recipient MDS mice compared to controls. Finally, donor chimerism of hematopoietic stem and progenitor cells (HSPCs) was higher in iron-restricted versus control MDS mice undergoing BMT. Conclusion: These findings demonstrate that pre-transplant conditioning profoundly alters iron homeostasis and restricting iron improves BMT outcome by increasing engraftment efficiency. Overall, these data suggest that conditioning-elicited NTBI is a critical mediator of peri-transplant toxicity and adversely impacts transplant outcome. Specifically, our observations prove that iron restriction improves donor HSPC engraftment in the recipient bone marrow, likely resulting in more efficient reconstitution of the HSPC pool and improved multilineage engraftment. Lastly, our results indicate that the peri-transplant application of novel iron restriction strategies aimed at reducing conditioning-induced NTBI - by positively affecting BMT outcome - is of potential benefit for patients undergoing BMT or gene therapy, presenting either with or without pre-transplant iron overload.

Identification and Validation of Ferroptosis-Related Genes As Novel Prognosis Prediction Panel for Acute Myeloid Leukemia

Acute myeloid leukemia (AML) is a main type of adult acute leukemia, especially in elderly population. The development of AML ascribed to various factors including chromosomal abnormalities, isolated gene mutations, etc. and the 5-year survival rate is around 30%. Currently, the most popular and precise assessment of AML is based on cytogenetic differences, and the patients with higher risk molecular mutations had inferior prognosis. However, with the development and application of new drugs, including bcl-2 inhibitors, many AML with high risks (such as FLT3, P53 mutated) also achieve better remission and longer survival. Up to date, many novel prognosis models are in process, with the better understanding of the initiation and development of leukemia. Ferroptosis, a widespread and ancient form of cell death driven by iron-dependent phospholipid peroxidation. Its involvement in AML has been reported, as leukemic cells frequently exhibit enhanced iron uptake and reduced iron efflux, resulting in elevated intracellular iron levels. Targeting iron homeostasis and inducing ferroptosis may provide novel insights into AML therapy. Herein, we integrated multi-gene information to identify a novel prediction model for ferroptosis-related genes (FRGs), and validated the accuracy of the model using clinical samples from our center (Figure A). In this study, we downloaded RNA-seq data and clinicopathological features of 151 AML patients from The Cancer Genome Atlas (TCGA) database (https://www.cancer.gov/ccg/research/genome-sequencing/tcga) and 70 corresponding health samples from the Genotype-Tissue Expression (GTEx) database (https://gtexportal.org/). Retrieve the ferroptosis-related genes (FRGs) set from the FerrDb database (http://www.zhounan.org/ferrdb/current/). The difference analysis, LASSO regression and Cox regression were used to determine the FRGs signature to construct prognostic models (Figure B). We finally identified 10 FRGs signature (ACSF2, SOCS1, CDO1, MYB, LPIN1, DNAJB6, PSAT1, GPX4, MT1G, FH) to establish a risk scoring model, and AML patients were divided into high-risk and low-risk group according to the median risk score. The risk score for each sample was determined using the following formula: . Kaplan-meier analysis showed that the overall survival of patients in the high-risk group was significantly worse( P<0.001) (Figure C). Cox regression analysis, ROC and DCA results found that these features have good predictive ability for prognosis of AML (Figure D-F). We also grouped the TCGA AML patients into poor/adverse and favorable/intermediate group based on NCCN guideline (risk stratification based on genomic mutation background), and found that the survival differences based on classical risk model were not that obvious ( P=0.062) (Figure G) when compared to our model ( P<0.001 in Figure C). Which suggested that our model may have its superior in distinguishing real high risk AML patients. In order to validate the accuracy of our model, we also conducted a retrospective study on 30 AML patients and 13 healthy controls (bone marrow stem cell donor) from September 2019 to September 2022. Bone marrow mononuclear cells (BMNC) from our biobank were obtained, and the ten FRGs panel was applied. Results indicated that there was a significantly difference between normal control and AMLs in FRGs expression (Figure H), and the survival data of high-risk AML patients with higher FRGs level was significantly inferior than that in low-risk group as expected ( P<0.05) (Figure I). To search for possible therapeutic drugs for high-risk patients, we downloaded AMLs data from the GDSC database (https://www.cancerrxgene.org/) and calculated the IC50 of commonly used chemotherapeutic drugs. We found that bcl-2 inhibitor (Obatoclax Mesylate) and TKI (Gefitinib) may have less sensitivity in our high-risk group ( P<0.05), while some unconventional drugs (paclitaxe, dactinomycin, camptothecin, irinotecan and vinorelbine) may benefit in patients with higher FRGs expression (Figure J). In summary, we identified and validated 10 FRGs signature to well predict the prognosis and drug sensitivity of AML. Based on our findings, appropriate combination of new drugs at the time of treatment may achieve unexpected therapeutic outcomes, but still need to go through further verification.

Ferroptosis in Parkinson's disease: Molecular mechanisms and therapeutic potential.

Parkinson's Disease (PD) is characterized by the progressive loss of dopaminergic neurons in the substantia nigra (SN), leading to motor and non-motor symptoms. While the exact mechanisms remain complex and multifaceted, several molecular pathways have been implicated in PD pathology, including accumulation of misfolded proteins, impaired mitochondrial function, oxidative stress, inflammation, elevated iron levels, etc. Overall, PD's molecular mechanisms involve a complex interplay between genetic, environmental, and cellular factors that disrupt cellular homeostasis, and ultimately lead to the degeneration of dopaminergic neurons. Recently, emerging evidence highlights ferroptosis, an iron-dependent non-apoptotic cell death process, as a pivotal player in the advancement of PD. Notably, oligomeric α-synuclein (α-syn) generates reactive oxygen species (ROS) and lipid peroxides within cellular membranes, potentially triggering ferroptosis. The loss of dopamine, a hallmark of PD, could predispose neurons to ferroptotic vulnerability. This unique form of cell demise unveils fresh insights into PD pathogenesis, necessitating an exploration of the molecular intricacies connecting ferroptosis and PD progression. In this review, the molecular and regulatory mechanisms of ferroptosis and their connection with the pathological processes of PD have been systematically summarized. Furthermore, the features of ferroptosis in PD animal models and clinical trials targeting ferroptosis as a therapeutic approach in PD patients' management are scrutinized.

Potential of Edible Seaweed of the Kenyan Coast as a Micronutrient Source

Edible seaweed, a rich source of micronutrients, remains underutilized despite its availability at the Kenyan coast, while micronutrient deficiencies remain a major health concern in Kenya. The study assessed micronutrient content of randomly sampled seaweed and consumption patterns, interviewing 175 seaweed farmers and analysing 58 samples from Mombasa, Kilifi, and Kwale counties. Micronutrients (vitamin A, D, B9, B12, iron, zinc, iodine) and heavy metals (mercury, lead, arsenic, cadmium) were measured. Predominantly female seaweed farmers, aged 30-40, with low income and education levels, focused on commercial farming over personal consumption due to limited nutritional awareness. A diverse array of edible seaweed species was identified; 11 Phaeophyceae (Sargassum, Turninaria, Dictyota, Padina, Eucheuma), 9 Chlorophyta (Ulva, Caulerpa, Eucheuma) and 8 Rhodophyta (Gracilaria, Hypnea, Soliera, Eucheuma). Significant regional and species variations were observed; Kilifi samples had higher vitamin A (p= 0.031), D (p=0.047), B9 (p=0.006), Kwale exhibited elevated iron levels (p= 0.047). Green phyla showed higher vitamin B12, D, zinc, iodine levels, while red phyla were rich in vitamin B9, A, and iron. Eucheuma cotonii had the highest vitamin B12 (696.6 μg/100 g) and vitamin A (298.6 μg/100 g) levels, Ulva intestinalis recorded highest iron (637.85 mg/kg) and Caulerpa taxifolia had highest zinc (277.74 mg/kg). Though some species contained traces of lead and cadmium within permissible limits, mercury was found in only four species, with no arsenic detected. Increasing awareness of seaweed's nutritional value is crucial to combatting micronutrient deficiencies, alongside further research into processing, value addition, market integration, and heavy metal mitigation strategies. Keywords; Edible seaweed, Micronutrients, Micronutrient deficiencies, Kenyan coast