Background. Research and clinical observation confirm a significant correlation between postpartum metabolic/obstetric diseases and the development of orthopedic pathology in highly productive cows. In particular, a study supported by the Russian Science Foundation (grant No. 24-26-00172) revealed a clear relationship between the development of purulent-necrotic lesions of the hooves and the occurrence of postpartum endometritis in high-yielding cows. In this study, the following groups of animals were formed by randomization: group 1 (n=28): cows with acute purulent-catarrhal postpartum endometritis; group 2 (n=25): animals with purulent-necrotic lesions of the hooves; group 3 (n=27): cows with a comorbid course of both pathologies; and the control group (n=23): clinically healthy animals. All individuals selected for the study had their jugular venous blood collected on an empty stomach into sterile tubes for subsequent immunological analysis. It was found that the comorbid course of these pathologies is significantly more severe than each of the diseases in isolation. This is evidenced by the immunological screening we conducted. In this regard, when an animal has multiple pathologies, an individual approach is necessary, which dictates the need for a comprehensive study of the clinical course of the main, concomitant, and even previous diseases, their comprehensive diagnosis, and rational treatment. Purpose. The purpose of the present paper is to study the immunological status of highly productive cows with comorbid obstetric-gynecological and orthopedic pathology. Materials and methods. To assess the dynamics of the clinical manifestation of hoof diseases, an orthopedic examination was conducted on a monthly basis throughout the year. During this examination, the following were evaluated: the degree and nature of hoof deformities; the intensity of corneal destruction; the presence of specific lesions such as pockets, delamination, and cracks; and the position of the thoracic and pelvic limbs. In cows with identified purulent-necrotic lesions during the postpartum period, a detailed obstetric and gynecological examination was additionally performed. It included a clinical examination, transrectal palpation, and ultrasound scanning, which was performed using the Scanner Falco device (8 MHz). Ultrasound was used to determine the size, echodensity, tissue homogeneity, and echogenicity of the reproductive structures. For immunological studies, blood was taken from the jugular vein of sick animals and cows from the control group (n=23) in the morning before feeding into sterile tubes. The total protein content in the blood serum of cows was determined using the biuret reaction, and the protein fractions were determined using the nephelometric method, and the A/G ratio was calculated. The amount of C-reactive protein was determined using the immunofluorescence method. The concentration of ceruloplasmin (CP) in the blood serum was determined by the standard method based on the oxidation of p-phenylenediamine with the participation of CP. The level of fibrinogen was determined spectrophotometrically, haptoglobin – by turbidimetric method. The level of circulating immune complexes (CIC) and their fractional composition were analyzed on the basis of determination of their molecular weight. The concentrations of interleukins (IL-1a and IL-8) and tumor necrosis factor-α (TNF-α) were measured by solid-phase enzyme immunoassay using commercial kits of monoclonal antibodies and reagents manufactured by Cytokin LLC, Saint Petersburg, Russia. Results. According to the Tukey multiple comparison analysis, the most significant increase in the level of CIC was observed in the group of animals with comorbid pathology. This indicates that the combined course of endometritis and hoof diseases leads to a more severe systemic inflammatory response compared to isolated forms of the diseases. This is evidenced by the indicators of protein metabolism, the level of acute phase proteins, the cytokine profile, and the amount of circulating immune complexes in the blood samples of the experimental animals. Conclusion. Currently, there is an urgent need for a comprehensive fundamental study of comorbidity in veterinary practice. The results of our research demonstrate that the comorbid course of orthopedic and obstetric-gynecological pathologies in high-yielding animals is characterized by a more severe clinical picture compared to isolated forms of monopathologies. This is evidenced by our immunological screening. In this regard, when an animal has multiple pathologies, an individual approach is necessary, which requires a comprehensive study of the clinical course of the main, concomitant, and even previous diseases, as well as their comprehensive diagnosis and rational treatment. In the future, it is necessary to create a universal tool that allows veterinarians to instantly and easily assess the structure, severity, and possible consequences of concomitant diseases in animals, conduct targeted diagnostics, and then prescribe the correct set of medical measures. Sponsorship information. The study was supported by the grant of the Russian Science Foundation No. 24-26-00172, https://rscf.ru/project/24-26-00172/. EDN: EQCWUC

Multiple sclerosis (MS) is a chronic neurodegenerative disorder for which dysregulated ferroptosis and necroptosis have demonstrated pathological associations but these lack causal validation in disease susceptibility. This study employed proteome-wide Mendelian randomization (MR) to investigate causal links between ferroptosis/necroptosis pathways, their upstream regulators, immune interactions, and MS risk. Transcriptomic validation utilized bulk RNA-seq and single-cell RNA-seq data. MR identified IFNA4 (OR = 0.24) and TNFAIP3 (OR = 2.0) as key causal ferroptosis/necroptosis-related proteins for MS risk. Analysis revealed 15 upstream regulators significantly associated with MS (FDR < 0.05; e.g., GZMA, CXCL3, APOE, CFB, CA6, KIR2DL2/3). Transcriptomic validation consistently identified ceruloplasmin (CP) as upregulated in MS microglia and lesions. Mediation analyses established two complete causal pathways: an IFNA4-mediated pathway wherein five upstream immune regulators (KIR2DL2, KIR2DL3, CFB, GZMA, and CA6) influence MS susceptibility through IFNA4 regulation, with all component effects statistically significant; and an APOE-driven pathway operating via TNFAIP3, demonstrating significant total effects and near-significant mediator-outcome effects on MS risk. While 59 immune traits were MS-associated, only TNFAIP3 showed a suggestive association with CD27⁺ memory B cells. This study establishes ferroptosis/necroptosis pathways as causal drivers of MS susceptibility, highlighting TNFAIP3, IFNA4, CP, and APOE as therapeutically actionable targets.

PurposeWilson’s Disease (WD), an autosomal recessive ATP7B mutations disorder causing copper accumulation, poses diagnostic challenges. This study used proteomics and single-cell transcriptomics to identify WD mechanisms and therapeutic targets.MethodsProteomic analysis was conducted on clinical samples from WD patients and the control group, followed by validation via ELISA. Subsequently, an integrated analysis was conducted by combining these data with single-cell RNA sequencing data from the database. Analytical content included differential expression, functional enrichment, drug target prediction, immune infiltration, and subtype-specific biomarker screening via LASSO/SVM-REF.ResultsProteomic analysis identified 420 differentially expressed proteins (266 upregulated, 154 downregulated) in WD patients compared with healthy controls, with significant enrichment in inflammatory pathways. Integration with DrugBank revealed eight hub proteins with high diagnostic accuracy (AUC > 0.9), among which Inter-alpha-trypsin inhibitor heavy chain 1 (ITIH1) and Transthyretin (TTR) may regulate the PI3K-Akt signaling pathway. Subsequently, ELISA validation confirmed significantly reduced levels of TTR, Ceruloplasmin (CP), and ITIH1 proteins in WD. Considering the heterogeneity of the WD microenvironment and single-cell diversity, further single-cell transcriptomic analysis was performed. The results revealed immune dysregulation, characterized by increased macrophage infiltration and reduced T/NK cell proportions, and PI3K-Akt-mTOR pathway enrichment in macrophages. For subtype-specific analysis, six key proteins were identified to distinguish hepatic and brain subtypes (AUC > 0.9).ConclusionsThe hub proteins and subtype-specific biomarkers identified in this study provide potential targets for the precise treatment of WD, while emphasizing the critical role of the PI3K-Akt pathway in WD.

The objective of the study was to determine the effects of prenatal auditory stimulation (PAS) on serum levels of heat shock protein 70 (HSP70), ceruloplasmin (CPN), alpha-1acid-glycoprotein (AGP), corticosterone (CORT), ovotransferrin (OVT), and dopamine (DA) in duck embryos and hatchlings. Fertilised Khaki Campbell duck eggs were subjected to the following auditory stimulation treatments: 1) no additional sound treatment other than the background sound of the incubator's compressors at 40 dB (CONTROL), 2) exposure to pre-recorded traffic noise at 90 dB (NOISE), and 3) exposure to Mozart's Sonata for Two Pianos in D Major, K 488 at 90 dB (MUSIC). The NOISE and MUSIC treatments were for 20 min/h for 24 h (a total of 8 h/d), starting from embryonic days (ED) 13 to hatching. Prenatal auditory stimulation did not influence hatchability rate, body weights at ED 21 and post-hatch day (PH) 1. The MUSIC and NOISE treatments elevated HSP70 at ED21 compared to the Control, while opposite results were observed at PH1. The AGP, CPN and CORT were not significantly (p > 0.05) affected by PAS at ED21. However, at PH1, the Control ducklings showed significantly higher OVT and AGP. The NOISE and MUSIC treatments attenuated DA activity at ED21 and PH1, respectively. PAS induces a physiological stress response in embryos but does not impact hatchability or hatchling weights. Exposure to MUSIC and NOISE effectively reduces stress during incubation and hatching, enhancing stress resilience in day-old ducklings.

Background/Objectives: The lifelong treatment of Wilson’s disease (WD) currently relies on copper chelators with relatively poor metal specificity, which frequently exhibit serious adverse effects. There is a real medical need for a specific copper chelator to regulate the copper excess efficiently, at lower doses than those used for penicillamine (DPA) or trientine (TETA), and with lower toxicity in long-term treatments. Methods: The efficiency of the specific Cu(II) chelator named TDMQ20 was evaluated by oral treatment of TX mice, used as a WD model, and compared with those of DPA, TETA, and also tetrathiomolybdate (bcTTM). We documented TDMQ20′s ability to (i) decrease the hepatic copper load, (ii) increase the amount and ferroxidase activity of ceruloplasmin (CP), and (iii) regulate liver proteins that are impaired in WD mice. Results: Compared to the other copper chelators, TDMQ20 was the only one that efficiently mediated excretion of Cu and restoration of active ceruloplasmin levels at doses 8 times lower than DPA. Such efficacy is related to the design of this chelator, which specifically coordinates Cu(II) as a discrete and soluble complex. Conversely, DPA, TETA, and bcTTM give rise to various complexes with copper ions, often with oligomeric or cluster structures that can be retained in blood circulation or sequestered by proteins. Conclusions: Taking into consideration all the advantages of TDMQ20 compared to other ligands, including its lack of toxicity during long-term administration in mice, the drug candidate TDMQ20 appears to be a first-class challenger to the currently used treatments, i.e., DPA, TETA, and bcTTM.

BackgroundArginase and ceruloplasmin are enzymes of redox balance involved in the metabolism of nitric oxide. Arginase competes with nitric oxide synthase (NOS) for L-arginine and hence plays a crucial role in the arginase/NOS balance of maintaining proteins and the appropriate nitric oxide (NO•) level in the serum. On the other hand, ceruloplasmin (CP) is an acute- phase protein responsible for the metabolic balance of copper and iron. For this study it was to investigate the serum concentrations of enzymes involved in the redox balance, namely arginase type I (Arg1) and CP, in a group of patients with and without sleep bruxism (SB), which was diagnosed by polysomnographic examination.Methods75 adults (35 women and 40 men, mean age 49.12) underwent a full- night of video polysomnography according to standards set out by the American Academy of Sleep Medicine. The concentration of Arg1 and CP in the serum, was determined using ELISA Kits.ResultsThe results showed that the concentration of Arg1 and CP was significantly lower in individuals with SB, irrespective of bruxism severity. Regression analysis revealed that only in the group of patients with higher Arg1 and CP concentrations, was there a negative linear relationship with the bruxism episode index (BEI).ConclusionThe results suggest that there is an oxidative imbalance in patients with SB, independent of the severity of bruxism. Higher plasma levels of Arg1 and CP were related to a lower BEI, potentially as the result of a protective biochemical balance against oxidative stress and inflammation in the SB.

ObjectiveIn this study, we retrospectively analyzed the relationship between PRL level and serum inflammatory and immune markers in patients with granulomatous lobular mastitis (GLM) and analyzed the effect of bromocriptine treatment on serum inflammatory and immune markers in patients with GLM. These analyses were conducted to illustrate that PRL is not only an endocrine hormone but also an immune factor, thereby providing evidence that GLM is an autoimmune disease.MethodsWe conducted a retrospective analysis of GLM cases admitted between 2023 and 2024. Clinical features were compared between patients with differential prolactin (PRL) levels using nonparametric tests, with concomitant documentation of prevalent clinical manifestations. Spearman’s rank correlation was employed to assess associations between PRL concentrations and clinical characteristics/serum biomarkers. To evaluate bromocriptine’s therapeutic efficacy, a propensity score-matched (PSM) cohort was established. Longitudinal serological changes were analyzed using nonparametric statistical methods for paired comparisons.ResultsElevated prolactin levels significantly correlated with lesion size (p<0.05). Patients with abnormal PRL exhibited lower 6-month cure rates compared to those with normal levels (93.1% vs. 100%, p=0.02). Baseline-PRL positively associated with neutrophil counts (NE#), Immunoglobulin E (IgE), and ceruloplasmin (CER) (all p<0.05). After treatment, baseline-PRL remained linked to elevated neutrophils, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), T4 lymphocyte percentage (CD4+ T cells), and IgE, but inversely correlated with lymphocytes (all p<0.05). Propensity-matched analysis (17 bromocriptine-treated vs. 13 no-treated) revealed reduced ESR, downregulated major histocompatibility complex class II (MHC-II) expression, and increased interleukin 4 (IL4), interleukin 5 (IL5), and regulatory T cell percentage (Treg%) levels in the treatment group (all p<0.05).ConclusionsOur findings suggest prolactin may act as an immunomodulatory factor in GLM, potentially influencing T/B-cell immunity and inflammatory cytokine recruitment. Additionally, the observed correlation between prolactin and ceruloplasmin positions ceruloplasmin as a candidate biomarker for GLM, though further validation in independent cohorts is required.

Metallothioneins (MTs) are a cysteine-rich protein that scavenges reactive oxygen species. Hypoxia is involved in the progression of diabetic nephropathy (DN) and aggravates oxidative stress. Hypoxia dramatically induced MT3, whereas induced around twofold increment in MT2, but inhibited MT1 in human renal proximal tubular epithelial cells (HRPTECs). Given that the role of MT3 in DN remains unclear, we explored the involvement of MT3 in DN. Microarray analysis also identified MT3-regulated candidate genes, including ceruloplasmin (CP) and cytochrome b reductase 1 (CYBRD1), as well as FGF-Klotho (KL)-FGFR complexes in HRPTECs. Hypoxia significantly induced MT3 expression through HIF-1-dependent mechanisms, and MT3 small interfering RNA (siRNA) decreased CP, CYBRD1, and KL expression under hypoxic conditions. In humanized MT3-BACTg mice, except HIF-1α, diabetes significantly increased the expression of MT3, CP, CYRBD1, FGFR2, and KL in the renal cortex in MT3-BACTg mice. Diabetic MT3-BACTg mice presented more severely damaged mitochondria in proximal tubules than their wild-type littermates did, accompanied with peritubular capillary obstruction by swollen endothelial cells. Moreover, the proximal tubules-specific overexpression of MT3 in mice (MT3Tg) represented no overlap in the protein expression between MT3 and HIF-1α in diabetic kidney. Accordingly, MT3 siRNA significantly augmented HIF-1α protein and HIF1A in HRPTECs. Finally, MT3 expression in the renal tubulointerstitium was positively correlated with the glomerular filtration rate (GFR) in DN subjects by data from Nephroseq. In conclusion, these results showed that there might be a unique interplay between MT3 and HIF-1α in diabetic kidney of to regulate hypoxia-induced HIF-1α expression.Supplementary InformationThe online version contains supplementary material available at 10.1007/s13340-025-00840-y.

ObjectiveExecutive dysfunction is a widespread and complex manifestation in schizophrenia, significantly impairing patients’ cognitive and functional outcomes. Despite extensive research, specific biomarkers associated with this dysfunction remain unidentified. Serum ceruloplasmin (Cp), a copper-binding protein involved in iron metabolism and oxidative stress regulation, has recently been implicated in neurological conditions. This study aims to investigate the relationship between reduced serum Cp levels and executive dysfunction in hospitalized schizophrenia patients, providing insights into potential biomarkers and therapeutic targets.MethodsA total of 95 schizophrenia inpatients treated at Shaoxing Seventh People’s Hospital from January 2023 to December 2024 were enrolled. Patients were divided into two subgroups based on serum Cp concentrations: 48 patients with reduced Cp levels (SC1 group, Cp < 200 mg/L) and 47 patients with normal Cp levels (SC2 group, 200-600 mg/L). Additionally, 42 age- and gender-matched healthy individuals served as the control group. Blood samples were collected for Cp measurement using an automated biochemical analyzer. The severity of psychiatric symptoms was assessed using the Positive and Negative Syndrome Scale (PANSS), while the Tower of London (TOL) test was employed to evaluate executive function. Statistical analyses included one-way ANOVA and Spearman’s correlation to examine group differences and relationships between Cp levels and cognitive performance.ResultsSignificant differences in serum Cp levels were observed among the SC1, SC2, and control groups (P < 0.01). Executive function, assessed via the TOL test, showed no significant difference between the SC1 and SC2 groups (P > 0.05); however, both groups exhibited significantly impaired performance compared to the control group (P < 0.001). A strong positive correlation was identified between Cp levels and TOL performance in the SC1 group (r = 0.890, P < 0.001), particularly in simpler and moderately complex tasks.ConclusionBased on the existing evidence of widespread cognitive impairment in schizophrenia patients, this study delves deeper into its potential causes. We found that regardless of whether schizophrenia patients had normal or low serum ceruloplasmin (CP) levels, their executive function was significantly lower than that of healthy individuals. Furthermore, a correlation analysis showed a significant link between CP levels and executive dysfunction in schizophrenia patients with decreased CP levels. Our study suggests that low CP levels may aggravate executive dysfunction, indicating that CP deficiency might be a biological marker of executive dysfunction in schizophrenia.

Growing evidence indicates that ferroptosis is pivotal in the development and progression of ovarian cancer (OC). However, the function of EGLN2 in OC remains unclear. In this study, we observed that EGLN2 is expressed at low levels in OC tissues and is associated with a favorable prognosis in early-stage patients based on data from TCGA and GTEx public databases. Compared with those in normal ovarian epithelial cell lines, EGLN2 mRNA and protein levels were significantly lower in OC cell lines. In vitro functional experiments revealed that EGLN2 overexpression reduced proliferation, increased the intracellular levels of iron ions, reactive oxygen species, lipid peroxidation, and mitochondrial oxidative phosphorylation, and inhibited the Warburg effect. Mechanistically, EGLN2 inhibited the expression of HIF-1α, which binds to the promoter of ceruloplasmin (CP), reducing the proliferation of and inducing ferroptosis in OC cells. A subcutaneous xenograft model in nude mice demonstrated that EGLN2 overexpression inhibited HIF-1α, promoted ferroptosis, and inhibited OC cell growth. In summary, EGLN2 suppressed CP transcription and increased ferroptosis in OC cells. These findings provide new insights into OC development and open avenues for innovative therapies.

This pilot study explored whether the ceruloplasmin (CP) and ferritin (FT) levels in ocular fluids could serve as biomarkers for early neurodegenerative diseases (Alzheimer's, Parkinson's, and other dementias). CP and FT are known to modulate neurodegenerative tissue responses. We analysed aqueous and vitreous samples from 26 patients (8M/18F, aged 60-85) who were undergoing elective vitreoretinal (VR) surgery. Of these, 14 had idiopathic epiretinal membranes (ERMs), 6 had idiopathic macular holes (MH), and 6 were patients with Alzheimer's disease (AD) who presented with VR disorders (VRDs). CP, FT, and selected neuroinflammatory mediators such as interferon γ (IFN-γ), interleukin (IL-6), vascular endothelial growth factor (VEGF), nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) were quantified. Odds ratio analysis was applied to assess the CP/FT ratio's association with subretinal drusen. We found distinct CP and FT profiles in VRD samples. In aqueous fluid, the CP increased and the FT decreased in early-stage ERM, which reduced the CP/FT ratio. Similar patterns were observed in vitreous fluid. The CP levels correlated with the VEGF (aqueous), IL-4 (vitreous), NGF, and BDNF levels; FT correlated with IL-6 and NGF. A higher CP/FT ratio was associated with increased risk for neurodegenerative conditions. Our findings support the quantification of CP and FT in ocular fluids as a promising approach for identifying early neurodegenerative changes and suggest that the CP/FT ratio may be linked to drusen imaging and clinical neurodegenerative history.

Determination of the concentration of acute phase proteins in blood plasma is a promising approach to assessing the degree of inflammation. The aim of the work was to study the levels of C-reactive protein (CRP), α1-acid glycoprotein (AGP), and ceruloplasmin (CP) in patients with thermal trauma. The concentration of CRP, AGP, and CP was measured in 27 patients with thermal injury (mean age 49.2 ± 5.3 years) on the 1st and 7th days of treatment. It was shown that at the early stage of burn injury, the concentration of CRP increased significantly (by 6.9 times), while the levels of AGP and CP were slightly elevated. A positive correlation was found between CRP concentration and burn area (r = 0.54, p &lt; 0.05). On day 7, CRP and AGP remained elevated, while CP levels were normalized. The most informative indicator for assessing the dynamics of inflammation in burn trauma was CRP, while AGP may be useful for monitoring the prolonged inflammatory response. Thus, the analysis of acute phase proteins can be useful for monitoring inflammation and the effectiveness of treatment in patients with burns.

Background and aim of the studyCompliance is the most challenging aspect of long-term therapy in Wilson’s disease (WD). Evidence is presented that butyrylcholinesterase (CHE) can be used as a sensitive biomarker to detect compliance problems in long-term treated WD-patients.MethodsFor the present retrospective, monocentric study demographical and treatment related data of 108 WD-patients (who had been treated at the Clinic of Neurology of the university hospital in Düsseldorf (Germany) between 2/2005 and 1/2021) were extracted from the charts. These patients underwent 2003 therapy control visits. The present study focuses on the analysis of three parameters of copper metabolism (serum levels of ceruloplasmin (CER-S), copper (CU-S) and the 24 h-urinary copper excretion (24 h-UCU)) and the serum levels of CHE (CHE-S). A patient was classified to be non-compliant when in his charts at least 8 24 h-UCU-values were found, and all his 24 h-UCU-values were larger than 60 μg/d (N-COM8-group). A patient was classified to be compliant when at least one of at least 8 24 h-UCU-values was lower than or equal to 60 μg/d (COM8-group).ResultsCHE-S was significantly (p < 0.05) different between thus defined compliant or non-compliant patients. Neither CU-S nor CER-S nor calculated free (non-ceruloplasmin-bound) serum copper levels (NCC-S) were significantly different between the NCOM8- and COM8-group. Analysis of the area under the curve and sensitivity and specificity by means of ROC-curves underlined the sensitivity of CHE-S in contrast to the insensitivity of CU-S and CER-S to detect patients who had been classified as compliant/non-compliant on the basis of their 24 h-UCU-values.ConclusionWhen compliance of WD-patients is classified on the basis of their 24 h-urinary copper excretion CHE-S is more sensitive to detect problems of non-compliance than serum levels of copper, of non-ceruloplasmin bound free copper or ceruloplasmin. Therefore, CHE-S may be used as an easy to determine further biomarker for compliance assessment in long-term treatment of WD in addition to 24 h-UCU.

Absence of astrocytic ceruloplasmin reverses the senescence process with aging of learning and memory abilities.

Abstract Cancer-associated fibroblasts (CAFs) are key contributors to promote cancer progression in the lung tumor microenvironment via interactions with cancer cells and other cell types. In a 3-D culture system, we classified CAFs into two subgroups based on their motility, motile and static CAFs, and isolated these subgroups using the photoconvertible fluorescent protein Dendra2. RNA sequencing and subsequent analyses revealed the upregulation of hypoxia- and glycolysis-related genes in motile CAFs. Activation of HIF-1α increased CAF motility in both mono- and co-culture systems. Knockdown of HIF-1α downstream genes, ALDOA and LDHA, reduced CAF motility. Interestingly, HIF-1α activity and CAF motility were heightened when co-cultured with mesenchymal-like lung cancer cells. Mass spectrometry analysis identified ceruloplasmin (CP) as a secretory protein derived from mesenchymal-like lung cancer cells. Knockdown of CP in mesenchymal-like lung cancer cells attenuated CAF motility in co-culture systems and lung tumor metastasis in a mouse model. Collectively, these findings demonstrated that CAF motility is regulated by HIF-1α signaling, which is modulated by lung cancer cell-derived CP. Citation Format: Jihye Park, Sieun Lee, Jonathan M. Kurie, Young-Ho Ahn. Enhanced motility of cancer-associated fibroblasts is mediated by HIF-1α signaling in the lung tumor microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 201.

Acute respiratory distress syndrome (ARDS), a common cause of acute fatal respiratory, is characterized by severe inflammatory lung injury as well as hallmarks of increased pulmonary vascular permeability, neutrophil infiltration, and macrophage accumulation. Tree shrew, a squirrel-like small animal model, has been confirmed to have more similar traits to human ARDS with one-hit intratracheal instillation of LPS in our previous study. In this study, we characterized protein profile changes induced by intranasal LPS challenge in the tree shrew model through tandem mass tag (TMT)-based quantitative proteomics and type II alveolar epithelial cells through pathological analysis. In total, 4070 proteins (p < 0.05) were identified from lung tissues of the LPS-induced group and PBS group. Among the differential expression proteins (DEPs) detected by t-test (≥|1.5-fold|), 529 DEPs were identified, of which 304 were upregulated, and 225 were downregulated. The most important pathways involved in the process of ARDS had been identified by enrichment analysis: oxidative stress, apoptosis, inflammatory responses, and vascular endothelial injury. In addition, proteins have been reported in animal models or clinical patients also detail investigated for further analysis, such as ceruloplasmin (CP), hemopexin (HPX), sphingosine kinase 1 (SphK1), lactotransferrin (LTF), and myeloperoxidase (MPO) were upregulated in induced tissues and confirmed by western blot analysis. Overall, this study not only reveals a comprehensive proteomic analysis of the ARDS tree shrew model but also provides novel insights into multi-pathways responses induced by the LPS challenge of tree shrews. We highlight shared and unique proteomic changes in the lungs of ARDS tree shrews and identify novel pathways for acute lung injury, which may promote the model into basic research and translational research.

Currently, research on ferroptosis-related prognostic models for gastric cancer is limited, whereas traditional predictive models often have a narrow perspective and low accuracy. In this study, we systematically analyzed the expression patterns of ferroptosis-related genes in patients with gastric adenocarcinoma and evaluated their prognostic value. Using data from The Cancer Genome Atlas (TCGA) and the FerrDb database, we developed a ferroptosis-related prognostic risk model based on four genes: hydroxycarboxylic acid receptor 1 (HCAR1), branched-chain amino acid transaminase 1 (BCAT1), ceruloplasmin (CP), and dickkopf-1 (DKK1). This model demonstrated strong prognostic value and potential clinical relevance in stratifying gastric cancer patients by overall survival outcomes. ferroptosis-related prognostic risk model. Compared to traditional clinicopathological features, the risk score derived from this model exhibited superior predictive accuracy for overall survival in patients with gastric cancer and served as an independent prognostic factor. Functional enrichment analysis revealed that the risk score was primarily enriched for extracellular matrix-related pathways. Additionally, the risk score was significantly correlated with TME signature genes, immune checkpoint expression, and immune cell infiltration in stomach adenocarcinoma (STAD). Mechanistic studies revealed that HCAR1 is abnormally overexpressed in gastric cancer tissues and is associated with a poor prognosis. It exerted its effects by regulating the GPX4/SLC7A11 axis to inhibit lipid peroxidation and malondialdehyde accumulation, thereby obstructing ferroptosis. Experimental validation demonstrated that the downregulation of HCAR1 promoted ferroptosis and suppressed malignant tumor phenotypes, suggesting that both the gene and its associated risk model hold significant clinical value as potential therapeutic targets and prognostic biomarkers.

Aceruloplasminemia (ACP) is a rare recessive disease caused by loss of ceruloplasmin activity due to pathogenic variants in the ceruloplasmin (CP) gene. ACP causes iron accumulation in various organs, leading to neurodegeneration, anaemia, and diabetes. Estimating ACP prevalence is challenging, particularly as missense variants are not readily identified as pathogenic. Heterozygous missense variants likely to impact function were mapped in gnomAD and representative examples analysed for effects on CP activity. This knowledge was complemented by prediction of destabilizing effects of potentially pathogenic missense variants and integrated with loss-of-function mutations. Global ACP prevalence was predicted and compared with a more traditional method. Several as yet uncharacterised missense CP variants of pathogenic interest were identified by structure-function in-silico analysis. A representative subset was functionally validated, together with known ACP missense variants. Insights on the relative importance of copper ions coordinating centres in CP and its substrate specificity were discovered. Overall, a destabilizing effect was predicted for 130 missense CP variants. This information, integrated with known ACP missense and loss-of-function CP variants in gnomAD, allowed an estimation of ACP prevalence of 12.6/106. An alternative analysis based on minor allele frequency ≤0.01 resulted in an ACP prevalence as high as 8/106. These prevalence estimates for ACP are 20-25-fold higher than previously estimated and underscore the applicability of structure-function based analyses of real-world genetic variability to provide an alternative method for representing the frequency of rare disease variants. REACT-EU PON 2014-2021, Kedrion S.p.A.

IntroductionCoronary artery ectasia (CAE) is a vascular anomaly characterized by abnormal coronary artery dilation, often associated with endothelial dysfunction and inflammation. While CAE shares features with coronary artery disease (CAD), its independent pathophysiology remains unclear, particularly in cases without concurrent CAD.AimTo evaluate oxidative and antioxidant biomarker levels in patients with isolated CAE to understand their role in its pathogenesis.Material and methodsOur study was conducted involving 48 patients with isolated CAE and 32 controls with normal coronary angiograms. Oxidative stress markers, including total oxidative status (TOS), oxidative stress index (OSI), and lipid hydroperoxide (LOOH), were measured, alongside antioxidant markers such as paraoxonase-1 (PON1), ceruloplasmin (CP), free sulfhydryl (SH) groups, and total antioxidant status (TAS).ResultsCAE patients exhibited significantly higher levels of TOS (30.14 ±8.81 vs. 23.88 ±4.74 mmol H2O2 equiv./l, p = 0.004), OSI (3.21 ±1.12 vs. 2.43 ±0.53 arbitrary units, p < 0.001), and LOOH (11.95 ±2.88 vs. 10.13 ±1.66 µmol H2O2 equiv./l, p = 0.003). No significant differences were found in TAS, PON1, CP, or SH levels between groups (p > 0.05 for all). Logistic regression identified smoking, TOS, and high sensitivity C-reactive protein (hsCRP) as independent predictors of CAE.ConclusionsElevated oxidative stress markers, particularly TOS, OSI, and LOOH, indicate a heightened pro-oxidant state in CAE, while antioxidant defenses remain largely unaltered. These findings suggest that oxidative stress may contribute to CAE pathogenesis, emphasizing the need for therapies targeting oxidative imbalance.

Benzyl butyl phthalate promotes ferroptosis in Sertoli cells via disrupting ceruloplasmin-mediated iron balance.