Identification of Ferroptosis-Related Gene Prognostic Signature and HSF1 for Reversing Doxorubicin and Gemcitabine Resistance in Uterine Carcinosarcoma

Purpose. Iron metabolism and ferroptosis play crucial roles in the pathogenesis of cancer. In this study, we aim to study the role of ferroptosis-related genes (FRGs) in uterine carcinosarcoma (UCS) and identify potential target for UCS. Methods. Prognostic differentially expressed FRGs were identified of in the TCGA cohort. Integrated analysis, cox regression, and the least absolute shrinkage and selection operator (LASSO) methods of FRGs were performed to construct a multigene signature prognostic model. Moreover, a dataset from Gene Expression Omnibus (GEO) served as an external validation. HSF1 was knockdown in MES-SA and FU-MMT-1 cells, and cell viability, lipid ROS, and intracellular iron level were detected when combined with doxorubicin or gemcitabine. Result. Five FRGs were selected to construct a prognostic model of UCS. The group with high-risk signature score exhibited obviously lower overall survival (OS) than the group with low risk signature score in both TCGA and validated GEO cohorts. Multivariate Cox regression analysis further indicated that the risk score was an independent factor for the prognosis of UCS patients. The high-risk group of UCS has a higher sensitivity in the treatment of doxorubicin and gemcitabine. Knocking down of HSF1 in MES-SA and FU-MMT-1 cells was more sensitive to doxorubicin and gemcitabine via increasing ferroptosis. Conclusions. The five FRGs risk signature prognostic model having a superior and drug sensitivity predictive performance for OS in UCS, and HSF1 is a potential marker sensitive to doxorubicin and gemcitabine in UCS patients.

TMT Quantitative Proteomics Analysis Reveals the Effects of Transport Stress on Iron Metabolism in the Liver of Chicken

Abnormal iron metabolism can cause oxidative stress in broilers, and transport stress (TS) may potentially influence iron metabolism. However, the mechanisms by which TS affects iron metabolism are unclear. This study used quantitative proteome analysis based on tandem mass tag (TMT) to investigate the effects of TS on liver iron metabolism in broilers. Broilers (n = 24) reared under the same conditions were selected randomly into the transported group for 4 h (T2) and non-transported group (T1). Results showed that the serum iron level and total iron-binding capacity of broilers in the T2 were significantly higher than those in the T1 (p < 0.05). The liver iron content of broilers in the T2 (0.498 ± 0.058 mg·gprot−1) was significantly higher than that in the T1 (0.357 ± 0.035 mg·gprot−1), and the iron-stained sections showed that TS caused the enrichment of iron in the liver. We identified 1139 differentially expressed proteins (DEPs). Twelve DEPs associated with iron metabolism were identified, of which eight were up-regulated, and four were down-regulated in T2 compared with T1. Prediction of the protein interaction network for DEPs showed that FTH1, IREB2, and HEPH play vital roles in this network. The results provide new insights into the effects of TS on broilers’ liver iron metabolism.

Evaluation of Plasma Levels of Interleukin 6 and Iron Status of Football Players in a Nigerian University

To determine the levels of interleukin 6 (IL-6) and iron status of football players in Madonna University, Elele, Rivers State, Nigeria. A total number of 100 subjects were recruited for the study, comprising of 50 of football players before playing football (25 males and 25 females) and 50 of football players after playing football (25 males, 25 females) from Madonna University Nigeria, Elele Campus, Rivers State, Nigeria. The data obtained from the study were presented as Mean SD in tables and analysed using student t-test for parametric data using SPSS version 20. The level of significance was set at p<0.05. The results showed significant increase (p=0.004) in interleukin 6 (IL-6) of football players after playing compared to it before playing and no significant change (p=0.505) in the iron level of football players after playing compared to before playing respectively. The results also showed no significant change in interleukin 6 compared among football players based on gender and age groups and no significant change in the iron level of all the subjects. The study showed increase an in interleukin 6 (IL-6) of the football players after playing compared to the level before playing which shows that the physical activity increases the level of interleukin 6 and but has no effect on the iron level after football game.

The Effects of a Single Blood Donation on the Lipid Profile, Iron Storage and Enzymatic Antioxidants

Cardiovascular disease (CVD) is a global illness causing 31% of global mortality. Though many factors contribute to CVD, oxidative stress advances atherosclerosis through several complementary components, such as the initiation of lipid peroxidation by iron. Blood donation may decrease the risk of CVD due to reducing the iron level. Literature reported that blood donors have a lower risk of CVD, possibly due to the lower iron levels. Various effects of blood donation are involved in preventing type II diabetes. However, little is known of the exact mechanism of the benefits of blood donation. In this study, samples were collected from 33 healthy male participants pre- (1 day) and post-donation (1 day, 1, 2 and 3 weeks) and the effect of the blood donation on the iron, lipids and enzymatic antioxidants profiles were assessed. A repeated-measures ANOVA was used for comparing the quantitative variables between the visits. We found that the iron decreased significantly by week 1 (–25.3%). Ferritin decreased significantly at weeks 1, 2, and 3 (–26.3%, –40.3%, –36.7%, respectively). The superoxide dismutase increased significantly at post-donation day 1, weeks 1, 2, and 3 (17.9%,35.7%, 31.1%, 36.6%, respectively) and in correlation with time [r (165) = 0.50, P <0 .01]. Glutathione peroxide decreased significantly at week 1 (–25.0%). Glutathione reductase decreased significantly 1-day post donation (–5.7%) then increased over the next three weeks [r (165) = 0.3, P <0.01]. Finally, the lipids were significantly reduced 24 hours after the donation but not at week 1, 2 and 3. We conclude that blood donation, resulting in a lowered body iron concentration, is an effective way to increase superoxide dismutase and glutathione reductase, which prevent the initiation of lipid oxidation. Our results could be used to advocate for the benefits of blood donation. However, further studies are required to assess the role of blood donation in plaque formation and arteriosclerosis.

Haemoglobin, Serum Iron Level and Cardiovascular Status in Advanced Stages of Chronic Kidney Disease Patients admitted in A Tertiary Care Centre

Background: Anaemia due to advanced stages of chronic kidney disease increases morbidity of patients. Early detection and correction of anaemia may be helpful in preventing the progress of the disease & its cardiovascular outcomes. The objective of this study was to evaluate hemoglobin, serum iron level and cardiovascular status in advanced of CKD patients. Methods: This was a cross sectional observational study on 150 cases of diagnosed advanced stages (3B,4 &5) of CKD patients in indoor of department of medicine of Mitford Hospital, Dhaka from July 2019 to January 2020. Convenience sampling was done. Data were analyzed with SPSS 26. Results: Total number of patients were 150. Male were 93 (62%) and female were 57 (38%). The mean (±SD) age was 55.22 (±10.30) years (range 33 - 75 years). Among the study subjects 38% had history of blood transfusion, 60% had history of iron supplementation and 12% subjects received erythropoietin. Mean (±SD) haemoglobin level was 7.61 (±2.54) g/dl. Seventy percent of the study subjects had haemoglobin level < 9 g/dl and rest had ³9 g/dl. Significant difference was found in between these two groups (p = 0.036). In the present study, mean (±SD) serum iron level was 15.59 (±07.39) μmol/l. In 46% of the study subjects, iron level was 7.3μ mol/l and 52% had iron level between 7.3 to 23.6 μmole/ lit. Mean (± SD) ferritin level of the study subjects was 155.22 (±92.32) ng/ ml. In 58% of the study subjects ferritin level was < 100ng/ ml and 42% had >100 ng/ml. Significant difference was found in between these two groups (p = 0.041) (Table-IV). Ferritin level had significant positive relationship with blood transfusion, iron and erythropoietin supplementation on logistic regression analysis. Haemoglobin and serum ferritin level was positively correlated with eGFR of the study subjects. Statistical analysis showed significant relationship between eGFR with haemoglobin and serum ferritin. Forty six percent of the study subjects had Transferrin saturation (TSAT) level below 20%. Fifty four percent subjects had a TSAT level above 20%. Significant difference was found in between these two groups (p = 0.001). In correlation analysis, haemoglobin, serum ferritin and TSAT level in the study subjects had negative relationship with duration of CKD in years. Relationship of haemoglobin and TSAT level with duration of CKD was statistically significant. In correlation analysis, serum TIBC level had negative relationship with haemoglobin level which was statistically significant. Serum iron, ferritin and TSAT level in the study subjects were positively correlated with haemoglobin level. Relationship of haemoglobin with serum iron level and TSAT level was statistically significant. Twenty percent of the study subjects had peripheral vascular disease, 111 (74%) of the study subjects had hypertension, 66 (44%) had ischaemic heart disease, 27 (18%) had a history of acute myocardial infarction, 24 (16%) had chronic heart failure, 93 (62%) had dyslipidemia and 27 (18%) patients had history of stroke. Conclusion: In this study it was observed that TSAT appears to be a more useful indicator for measuring the frequency of iron deficiency than serum iron, TIBC and serum ferritin. The cardiovascular comorbidities plagued significant number of patients with advanced CKD. Bangladesh J Medicine July 2022; 33(1) : 34-39

Non-suicidal intentional ingestion of iron tablets and severe intoxication: the result of adolescent boy’s impulsive risky behaviour in the school

Acute iron poisoning is a potentially fatal intoxication in children. As the Iron preparations are commonly administered to pregnant women, lactating mothers, toddlers, it is easily available at home. So younger children are prone to consume it accidently. Although iron is a therapeutic drug in recommended dosages, excessive iron in the free state can produce toxicity by affecting multiple cellular processes by catalysing redox reactions with lipid peroxidation and free radical formation. The severity of intoxication depends on the amount of elemental iron ingested. Serious toxicity is usually associated with a dose of >40 mg/kg of elemental iron. Levels more than 100 mg/kg are almost always fatal. We report a case where a 12-year male child intentionally taken 60 tablets of iron (ferrous fumarate) at his school as a part of competition or bet to other schoolmate and presented with acute iron poisoning with hepatic encephalopathy to us. Important initial laboratory parameters were AST-4,879 U/L, Prothrombin time-60 sec and Iron level-213 microgram/dl. With timely specific management i.e., deferoxamine infusion along with all required intensive care supportive management in PICU the patient was discharged successfully. We chose to report this case to highlight the risky behaviour of adolescence who usually grows physically and emotionally earlier but their prefrontal lobes are yet immature to take proper and correct decision. Thus, impulse activity may prove fatal for them.

Iron, Heme Synthesis and Erythropoietic Porphyrias: A Complex Interplay

Erythropoietic porphyrias are caused by enzymatic dysfunctions in the heme biosynthetic pathway, resulting in porphyrins accumulation in red blood cells. The porphyrins deposition in tissues, including the skin, leads to photosensitivity that is present in all erythropoietic porphyrias. In the bone marrow, heme synthesis is mainly controlled by intracellular labile iron by post-transcriptional regulation: translation of ALAS2 mRNA, the first and rate-limiting enzyme of the pathway, is inhibited when iron availability is low. Moreover, it has been shown that the expression of ferrochelatase (FECH, an iron-sulfur cluster enzyme that inserts iron into protoporphyrin IX to form heme), is regulated by intracellular iron level. Accordingly, there is accumulating evidence that iron status can mitigate disease expression in patients with erythropoietic porphyrias. This article will review the available clinical data on how iron status can modify the symptoms of erythropoietic porphyrias. We will then review the modulation of heme biosynthesis pathway by iron availability in the erythron and its role in erythropoietic porphyrias physiopathology. Finally, we will summarize what is known of FECH interactions with other proteins involved in iron metabolism in the mitochondria.

A low serum iron level is a potential predictor of poor renal function in patients following laparoscopic sleeve gastrectomy: a retrospective study

This study aimed to assess the association of serum iron level (Iron) with the estimated glomerular filtration rate (eGFR) after bariatric surgery (BS). We reviewed 210 patients with mean age of 39.1 ± 10.6 years (body mass index, 41.4 ± 5.5 kg/m2) undergoing BS. The primary outcome was the relationship between Iron and eGFR at 12-month after surgery. Multiple linear regression analyses were performed using postoperative eGFR as dependent variables and using Iron and other variables (i.e., age) as independent variables. At 12-month follow-up, 94 patients were analyzed. BMI significantly decreased, whereas serum iron level significantly increased. Although the percentage of patients with eGFR of < 90 mL/min/1.73 m2 increased during the study period, no significant difference was found in postoperative 12-month eGFR. No correlations were noted between Iron and eGFR at baseline and postoperative 1 and 6 months, whereas a significant relationship was observed between Iron and postoperative 12-month eGFR. Multiple linear regression analyses revealed that Iron and presence of diabetes were the independent predictors of postoperative 12-month eGFR. This pilot study showed a positive association of postoperative serum iron level with renal function in this patient population. Further large-scale trials are needed to confirm the findings.

Evaluation of Serum and Salivary Iron and Ferritin Levels in Children with Dental Caries: A Meta-Analysis and Trial Sequential Analysis

Background and objective: Dental caries appears to be related to iron deficiency anemia and to low ferritin levels. In the present meta-analysis, we report salivary and serum iron and ferritin levels in children with dental caries, compared to healthy controls. Materials and methods: We searched in Web of Science, Cochrane Library, Scopus, and PubMed/Medline databases to extract studies published until 25 July 2021. We calculated mean differences (MD) and 95% confidence intervals (CI) of salivary and serum iron and ferritin levels in children with dental caries, always compared to healthy controls. In addition, we applied a trial sequential analysis (TSA). Results: A total of twelve articles covering thirteen studies were included in the meta-analysis. The pooled MD for salivary iron level was −5.76 µg/dL (p = 0.57), and −27.70 µg/dL (p < 0.00001) for serum iron level: compared to healthy controls, children with dental caries did not show different salivary iron levels, while children with caries had significantly lower serum iron levels. The pooled MD of salivary ferritin level was 34.84 µg/dL (p = 0.28), and the pooled MD of serum ferritin level was −8.95 µg/L (p = 0.04): compared to healthy controls, children with dental caries did not have different salivary iron levels, but significantly lower serum ferritin levels. Conclusions: The findings of the present meta-analysis showed that salivary levels of iron and ferritin did not differ between children with and without caries, though compared to healthy controls, children with caries had significantly lower salivary and serum iron and ferritin levels. The results are of practical and clinical importance: Possibly, iron and ferritin supplementation might prevent or attenuate dental caries in children at risk. Further, children with caries might suffer from further iron- and ferritin-related health issues. Lastly, serum blood samples, but not saliva samples inform accurately about the current iron and ferritin concentrations in children with or without caries.

Treating Iron Overload Disorders with a Novel Therapeutic Antibody Targeting TMPRSS6

Iron is an essential element for almost all living organisms as it participates in a wide variety of metabolic processes. Disorders of iron metabolism are among the most prevalent human diseases, ranging from anemia to hemochromatosis. Excessive iron accumulations in major organs of iron overload patients can lead to high mortality. Hepcidin, a HAMP-encoded liver hormone, is the master regulator of iron homeostasis. By binding to the sole iron exporter ferroportin and causing internalization and degradation of the complex, hepcidin inhibits cellular iron efflux, thereby lowers plasma iron levels. Inappropriately suppressed/low hepcidin production is central to iron overload. Transmembrane protease serine-6 (TMPRSS6), a type II transmembrane serine protease primarily expressed in liver, downregulates hepcidin expression through BMP-SMAD pathway. TMPRSS6 deficiencies have been shown to cause hepcidin overexpression in both TMPRSS6-mutant mice and in patients with iron-refractory iron deficiency anemia (IRIDA). Therefore, TMPRSS6 is a viable therapeutic target for iron overload disorders. Here we report the generation of an anti-TMPRSS6 antibody through a hybridoma campaign using a DNA-based immunization approach, followed by humanization and sequence optimization. Lead antibody, hzMWTx-003 selectively binds human TMPRSS6 with low nanomolar affinity (KD: 7.6nM), and is cross-reactive to rodent (mouse and rat) and monkey (cynomolgus and rhesus) TMPRSS6. Single-dose injection of hzMWTx-003 was able to significantly elevate serum hepcidin and liver HAMP RNA levels in wildtype mice, resulting in significantly reduced serum iron level. The Hbb th3/+ mouse model of β-thalassemia, like its human counterpart, is characterized by iron overload, ineffective erythropoiesis and splenomegaly. Treatment of Hbb th3/+mice with MWTx-003 effectively increased hepcidin expression at both protein and RNA levels, leading to significantly reduced serum iron and liver non-heme iron content. MWTx-003 also dramatically improved anemia and ineffective erythropoiesis, and alleviated splenomegaly in these mice. CMC development of hzMWTx-003 confirms outstanding biophysical properties. Preliminary studies in cynomolgus monkey using GLP-grade material demonstrated good pharmacokinetics of hzMWTx-003 and expected pharmacodynamic response where reduction of serum iron could be sustained for 21 days after single dose administration. A single dose toxicology study in cynomolgus monkey revealed no safety concerns, and no production of anti-idiotype antibodies was detected. In summary, anti-TMPRSS6 antibody MWTx-003 represents a promising therapy for iron overload disorders such as β-thalassemia, and potentially other diseases where iron restriction is beneficial. Disclosures Chen: Mabwell Therapeutics Inc: Current Employment. Wang: Mabwell Therapeutics Inc: Current Employment. Zheng: Mabwell (Shanghai) Bioscience Co. Ltd: Current Employment. Huang: Mabwell Therapeutics Inc: Current Employment. Mao: Mabwell (Shanghai) Bioscience Co. Ltd.: Current Employment. Ouyang: Mabwell (Shanghai) Bioscience Co. Ltd.: Current Employment. Du: Mabwell Therapeutics Inc: Current Employment.