Biochimica et biophysica acta. General subjects最新文献

{"title":"Nanobody-based imaging: Advancing precision in molecular diagnostics","authors":"Zohre Eftekhari,&nbsp;Fatemeh Kazemi-Lomedasht","doi":"10.1016/j.bbagen.2025.130884","DOIUrl":"10.1016/j.bbagen.2025.130884","url":null,"abstract":"<div><div>Molecular imaging is a cornerstone of modern medicine, enabling non-invasive visualization of biological processes at the molecular level. The emergence of nanobodies (Nbs), small single-domain antibody fragments derived from camelids, has transformed this field due to their superior tissue penetration, rapid clearance, and high target specificity compared to conventional antibodies. This review focuses on the integration of Nbs with positron emission tomography (PET) and single-photon emission computed tomography (SPECT) two complementary molecular imaging modalities known for their high sensitivity, quantitative potential, and clinical relevance. Nb-based PET and SPECT imaging probes are emerging as powerful tools for detecting disease-associated molecular targets with exceptional precision. Their unique properties, including high affinity, specificity, and stability, make them ideal candidates for developing advanced radiotracers that enable early disease detection, monitoring of therapeutic responses, and evaluation of novel treatment strategies. Despite these advantages, several challenges remain, such as scalable Nb production, reduction of immunogenicity in clinical applications, and optimization of radiolabeling methods that preserve Nb integrity and function. This review highlights recent advances in Nb engineering, radiolabeling strategies, and preclinical and clinical applications of Nb-based PET and SPECT imaging, while outlining critical directions for future research. By addressing current limitations, Nb-based molecular imaging holds great promise for improving diagnostic accuracy, advancing personalized medicine, and expanding its clinical impact across diverse disease areas.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1870 1","pages":"Article 130884"},"PeriodicalIF":2.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145487720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selection and evaluation of new sites for splitting beta-lactamase to modulate auto-complementation enzyme activity","authors":"Morgan C. Marsh ,&nbsp;Shawn C. Owen","doi":"10.1016/j.bbagen.2025.130867","DOIUrl":"10.1016/j.bbagen.2025.130867","url":null,"abstract":"<div><div>Split protein systems, including split fluorescent proteins and split enzymes, have predominantly been applied to monitor protein-protein interactions but have recently gained popularity as components in induced complementation diagnostic platforms. Many of these systems are limited by self-affinity which limits the dynamics or causes a false-positive high background signal. To overcome these challenges, it is necessary to either modify the interacting residues of established split fragments or by exploring new split-sites. Using the computational modeling tool SPELL to identify stable candidates, we developed five unique split sites for TEM-1 β-lactamase to compare structure and activity versus the original split site. We monitored structural changes through MMS IR spectroscopy as well as functional activity of each split enzyme. Expected structural changes of each fragment and changes in self-affinity driven complementation were observed. By utilizing specific binding proteins attached to the split enzyme, we monitored enzyme activity with forced complementation when bound to the target compared to auto-complementation when the enzymes were auto-complexed in solution. Of the five new split sites, one maintained high binding-mediated complementation activity with significant reduction in background auto-complementation. Changing the split site is a relatively simple approach to reduce background activity while maintaining on-target activity. The ability to tune split-systems has the potential to enable more sensitive and complex studies that previously were limited by the high background from self-affinity.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1870 1","pages":"Article 130867"},"PeriodicalIF":2.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145428825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rationally designed peptide induces apoptosis and cell cycle modulation in resistant melanoma","authors":"Layza Sá Rocha ,&nbsp;Ana Cristina Jacobowski ,&nbsp;Eduarda Thiburcio ,&nbsp;Rafael Araujo Pereira ,&nbsp;Claudiane Vilharroel Almeida ,&nbsp;Camila De Oliveira Gutierrez ,&nbsp;Thaís de Andrade Farias Rodrigues ,&nbsp;Rodrigo Juliano Oliveira ,&nbsp;Gabriel B. Taveira ,&nbsp;Priscila Aiko Hiane ,&nbsp;Ana Paula de Araújo Boleti ,&nbsp;Octávio Luiz Franco ,&nbsp;Marlon Henrique Cardoso ,&nbsp;Maria Ligia Rodrigues Macedo","doi":"10.1016/j.bbagen.2025.130877","DOIUrl":"10.1016/j.bbagen.2025.130877","url":null,"abstract":"<div><div>The ineffectiveness of conventional therapies against melanoma necessitates the development of novel approaches characterized by high selectivity and low systemic toxicity. Using computational optimization, KW18, a synthetic α-helical peptide, was rationally engineered. It incorporates conserved bioactive motifs from patented peptides. The Joker algorithm was used to refine the peptide. This process enhanced its positive charge, hydrophobicity, and α-helical stability. In vitro, KW18 displayed potent cytotoxicity against melanoma cells (IC₅₀ = 11.41–21.75 μM) with 84–91 % inhibition at 128 μM, minimal toxicity to FN1 fibroblasts, hemolysis below 5 %, and stability exceeding 80 %. Functional assays revealed mitochondrial damage and caspase-dependent apoptosis (86.5 % late apoptosis at 24 h). Cell cycle analysis in B16F10-Nex2 cells revealed a moderate accumulation in G0/G1 phase (62.9 %) alongside a substantial S-phase population (25.8 %), with minimal Sub-G0 (5.2 %) and reduced G2/M (6.1 %). This profile suggests a cytostatic-like effect through partial cell cycle slowdown. Structural characterization via helical wheel projections and circular dichroism confirmed its α-helical conformation, consistent with membrane-targeting activity. In vivo, <em>Galleria mellonella</em> larvae exposed to KW18 exhibited 100 % survival, supporting a favorable safety profile. Collectively, KW18 combines apoptosis induction with cell cycle modulation, offering a dual mechanism against melanoma while sparing healthy cells. These findings designate KW18 as a stable, selective, and safe therapeutic option for drug-resistant melanoma and a beneficial element for combined treatments.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1870 1","pages":"Article 130877"},"PeriodicalIF":2.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145443628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The CEBPA/FDX1 axis elevates sensitivity to cuproptosis in lung adenocarcinoma cells","authors":"Yongsheng Zhao ,&nbsp;Renyan Zheng ,&nbsp;Kexin Luo ,&nbsp;Haiyang Zhao ,&nbsp;Wanping Xiang","doi":"10.1016/j.bbagen.2025.130872","DOIUrl":"10.1016/j.bbagen.2025.130872","url":null,"abstract":"<div><h3>Background</h3><div>The recent introduction of “cuproptosis” into the oncological lexicon has opened up new horizons for cancer therapy, yet our understanding of how this process operates in lung adenocarcinoma (LUAD) is still limited. In this work we have investigated the core genetic factors and provided theoretical support for the application of cuproptosis in the treatment of LUAD. Through bioinformatics analysis, we found that FDX1 was significantly downregulated in LUAD and highly correlated with cuproptosis markers. Therefore, we chose FDX1 as the research object.</div></div><div><h3>Methods</h3><div>Biochemical and bioinformatic analyses were assessed FDX1 and CEBPA expression in LUAD. The binding relationship between CEBPA and FDX1 was validated <em>via</em> CHIP and dual-luciferase assays. FDX1 expression in LUAD cells was evaluated by qRT-PCR and Western blot. The impact of FDX1 overexpression on LUAD progression was examined using CCK-8 and Transwell assays. Experiments involving CCK-8, copper ion measurement, cuproptosis-related protein detection, and DLAT immunofluorescence confirmed the successful LUAD cuproptosis model.</div></div><div><h3>Results</h3><div>FDX1 was downregulated in LUAD tissues and cells, and its overexpression inhibited LUAD cell migration, invasion, and proliferation. Cuproptosis significantly reduced LUAD cell viability and the protein levels of Lipoy-DLAT, DLAT, and FDX1, while increasing HSP70 expression, DLAT aggregation, and intracellular copper ion levels. CEBPA, a transcriptional activator of FDX1, positively correlated with and bound to it. Overexpressed FDX1 enhanced cuproptosis in LUAD cells, an effect partially reversible by CEBPA suppression.</div></div><div><h3>Conclusion</h3><div>These analyses were performed to construct a proposed cellular model of cuproptosis in LUAD.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1870 1","pages":"Article 130872"},"PeriodicalIF":2.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145367392","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Screening of biomarkers in autoimmune thyroid diseases (AITDs): preliminary study based on Raman spectroscopy and MALDI-ToF mass spectrometry","authors":"Sara Trzos ,&nbsp;Sylwia Orzechowska ,&nbsp;Paweł Link-Lenczowski ,&nbsp;Grzegorz Sokołowski ,&nbsp;Malgorzata Baranska ,&nbsp;Ewa Pocheć","doi":"10.1016/j.bbagen.2025.130883","DOIUrl":"10.1016/j.bbagen.2025.130883","url":null,"abstract":"<div><div>Hashimoto's thyroiditis (HT) and Graves' disease (GD), autoimmune thyroid diseases (AITDs), are among the most commonly reported autoimmune disorders. Early and unambiguous diagnosis and monitoring of the disease development are crucial to obtain the most effective treatment results. Matrix-assisted laser desorption/ionization with a time-of-flight analyzer mass spectrometry (MALDI-ToF MS) and Raman spectroscopy are promising analytical methods in this regard. These non-invasive and sensitive techniques provide information on specific features of proteins, including their post-translational modifications and the level of a wide range of biomolecules in the bloodstream, which are useful for assessing the health status of patients. This study screened for potential biomarkers in AITDs using both methods. Sera from HT patients at two stages of disease progression and GD patients before and after normalization of thyrotropic hormone following immunosuppressive treatment were used in the study. Serum biomolecule changes were analyzed using Raman spectroscopy. <em>N</em>-glycans released from serum glycoproteins were detected by MALDI-ToF mass spectrometry. We observed that the levels of phenylalanine, carotenoids, and phospholipids in HT sera correlate with increased inflammation accompanying this disease. The GD group showed a lower amount of serum collagen compared to the HT patients. Quantitative comparison of <em>N</em>-glycans revealed several differences between the study groups and healthy donors. The higher galactosylation and α2,6-sialylation of serum proteins in HT2 relative to GD patients are the main differences in <em>N</em>-glycan profiles of AITDs. The obtained preliminary results demonstrate that these analytical techniques have potential in diagnosing and monitoring AITDs, and can be a good alternative to the currently used methods. Analysis of a larger number of samples, as well as a more detailed MS methodology for precise decoding of glycan structures, is necessary for further research.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1870 1","pages":"Article 130883"},"PeriodicalIF":2.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145470321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hyperglycemia accelerated the metastasis of triple-negative breast cancer via promoting TNFα/Gli-1 axis in endothelial cells","authors":"Xiyu Mei ,&nbsp;Chuang Ke ,&nbsp;Ziyun Gao ,&nbsp;Fan Yang ,&nbsp;Zhenlin Huang ,&nbsp;Bin Lu ,&nbsp;Lili Ji","doi":"10.1016/j.bbagen.2025.130875","DOIUrl":"10.1016/j.bbagen.2025.130875","url":null,"abstract":"<div><div>Diabetes mellitus (DM) is associated with a poor prognosis of aggressive breast cancer. Vascular dysfunction is commonly found during the development of both cancer and diabetes. We previously reported that the disruption of vascular endothelial phenotype induced by tumor necrosis factor-<em>α</em> (TNF<em>α</em>) accelerated the trans-endothelial metastasis of triple-negative breast cancer (TNBC). Herein, we explored the role of vascular endothelial cells in diabetes-induced TNBC metastasis<em>.</em> Both type 2 DM (T2DM) and type 1 DM (T1DM) enhanced the metastasis of TNBC <em>in vivo</em>. T2DM increased the expression of endothelial phenotype vascular endothelial cadherin (VE-cadherin), platelet-endothelial cell adhesion molecule (PECAM-1/CD31), and mesenchymal markers including vimentin and fibroblast specific protein-1 (FSP-1/S100A4) in tumor vessels. T1DM increased the expression of vimentin and FSP-1, but suppressed the expression of VE-cadherin in tumor vessels. Hyperglycemia elevated the production of TNF<em>α in vivo</em> and <em>in vitro</em>. TNF<em>α</em> reduced the trans-endothelial electrical resistance (TEER) value of both human mammary microvascular endothelial cells (HMMECs) and human umbilical vein endothelial cells (HUVECs). Expressions of vimentin and <em>α</em>-smooth muscle actin (<em>α</em>-SMA) were also increased in TNF<em>α</em>-treated both HMMECs and HUVECs. The number of trans-endothelial migrated MDA-MB-231 cells through TNF<em>α</em>-treated HMMECs or HUVECs monolayer was elevated. Moreover, glioma-associated oncogene 1 (Gli-1) was remarkably accumulated in the nucleus of TNF<em>α</em>-stimulated HMMECs and DM-induced tumor vessels. Both Gli-1 siRNA and GANT61 (an inhibitor of Gli-1) could abrogate the increased TNBC trans-endothelial migration through TNF<em>α</em>-treated ECs. We demonstrated that DM might promote TNBC metastasis via activating the TNF<em>α</em>/Gli-1 axis initiated vascular endothelial mesenchymal-like phenotype.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1870 1","pages":"Article 130875"},"PeriodicalIF":2.2,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145437054","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"m1A methylation-mediated upregulation of RILsPL1 promotes colorectal cancer progression via the CaMKII/CREB signaling pathway","authors":"Songbo Yang ,&nbsp;Xianghuai Long ,&nbsp;Wuliang Diao ,&nbsp;Jiaxing Yang ,&nbsp;Anyuan Jiang ,&nbsp;Hongfei Wu","doi":"10.1016/j.bbagen.2025.130856","DOIUrl":"10.1016/j.bbagen.2025.130856","url":null,"abstract":"<div><div>Colorectal cancer (CRC) remains one of the most lethal malignancies globally, driven by complex molecular mechanisms that contribute to its progression and metastasis. This study focuses on the role of N1-methyladenosine (m¹A) RNA methylation in CRC, particularly its effect on Rab Interacting Lysosomal Protein-Like 1 (RILPL1) expression and the downstream activation of the CaMKII/CREB signaling pathway. Bioinformatics analysis identified RILPL1 as a key gene associated with poor CRC prognosis, exhibiting increased expression levels in cancerous tissues, with further elevation in metastatic samples. Functional assays demonstrated that m¹A methylation enhances the stability of RILPL1 mRNA, a process dynamically regulated by the opposing actions of the demethylase ALKBH1 and the methyltransferase TRMT6. Loss-of-function and gain-of-function studies showed that RILPL1 promotes CRC cell viability, invasion, and migration, highlighting its oncogenic role. <em>In vivo</em>, RILPL1 knockdown markedly suppressed tumor growth in a nude mouse xenograft model. Furthermore, the CaMKII/CREB signaling pathway was identified as a critical mediator, with RILPL1 expression levels directly correlating with the phosphorylation of CaMKII and CREB both <em>in vitro</em> and <em>in vivo</em> xenograft models. Pharmacological rescue experiments confirmed this dependency, as a CaMKII activator reversed the effects of RILPL1 knockdown, while a specific inhibitor blocked this rescue. These findings suggest that dynamic m¹A methylation-driven upregulation of RILPL1 contributes to CRC progression through the activation of the CaMKII/CREB signaling pathway, offering potential therapeutic targets for CRC treatment.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 12","pages":"Article 130856"},"PeriodicalIF":2.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145013811","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Seasonal and COVID-19 lockdown variations in PM2.5 composition to apoptosis pathways and chemical toxicity in lung cells","authors":"Ting-Hsuan Wu ,&nbsp;Yu-Cheng Chen ,&nbsp;Hong-Lin Chan ,&nbsp;Hsiu-Chuan Chou","doi":"10.1016/j.bbagen.2025.130864","DOIUrl":"10.1016/j.bbagen.2025.130864","url":null,"abstract":"<div><h3>Objective</h3><div>To investigate seasonal and COVID-19-lockdown variations in PM2.5 chemical composition in Taipei and the effects of those compositional differences on lung cell toxicity.</div></div><div><h3>Methods</h3><div>PM2.5 was collected in warm (2021/07–2021/08) and cold (2021/11–2022/02) seasons and during LV2/LV3 COVID-19 alert periods. Composition (metals, PAHs) was analyzed and A549 and PC9 lung cells were exposed to equal mass concentrations (0–100 μg/mL) of extracted PM2.5. Cell viability, ROS, apoptosis, mitochondrial membrane potential and GSH/GPX4 were measured.</div></div><div><h3>Results</h3><div>At equivalent mass concentrations (50 μg/mL), warm-season PM2.5 induced higher ROS and greater reduction in viability than cold-season PM2.5. LV2-period PM2.5 contained higher metal content and caused more severe cellular damage than LV3. Spearman correlation plus single-compound assays identified cadmium and Dibenzo[<em>a</em>,<em>e</em>]pyrene as components strongly associated with cytotoxicity.</div></div><div><h3>Conclusions</h3><div>Seasonal and human-activity driven compositional changes—rather than mass concentration alone—affect PM2.5 toxicity in vitro; these results underscore the importance of controlling specific chemical emissions.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 12","pages":"Article 130864"},"PeriodicalIF":2.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145237919","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Untargeted metabolomics and proteomics reveal the versatile effects of myeloperoxidase-oxidized LDL on endothelial cells","authors":"Cecilia Tangeten ,&nbsp;Axelle Bourez ,&nbsp;Alexandre Rousseau ,&nbsp;Virginie Imbault ,&nbsp;Jianru Stahl-Zeng ,&nbsp;Florence Souard ,&nbsp;Xavier Bisteau ,&nbsp;Cedric Delporte ,&nbsp;Karim Zouaoui Boudjeltia ,&nbsp;Pierre Van Antwerpen","doi":"10.1016/j.bbagen.2025.130865","DOIUrl":"10.1016/j.bbagen.2025.130865","url":null,"abstract":"<div><h3>Background</h3><div>Myeloperoxidase-oxidized LDLs (Mox-LDLs) trigger endothelial cells and contribute to the development of atherosclerosis. However, the mechanisms underlying Mox-LDL-induced stimulation remain elusive. In this study, we applied untargeted metabolomics and proteomics approaches to investigate human umbilical vein endothelial cells (HUVECs) following exposure to Mox-LDLs.</div></div><div><h3>Methods</h3><div>HUVECs were exposed for 24 h to Mox-LDLs (0 or 100 μg/ml) with or without native LDLs (0 or 1 mg/ml). Supernatants and cell lysates were analyzed by liquid chromatography coupled to mass spectrometry. Using Workflow4Metabolomics work environment, MZmine, SIRIUS and MetGem, we selected and identified key metabolites influenced by Mox-LDL treatment. For the proteomics analysis, we used DIA-NN and the FragPipe-Analyst application to detect proteins differentially expressed after Mox-LDL treatment.</div></div><div><h3>Results</h3><div>Metabolomics analysis revealed increased levels of sphingolipids, phospholipids and oxidized-cholesterol derived compounds in HUVECs following Mox-LDL exposure. We also detected an increase in small peptides, likely reflecting Mox-LDLs catabolism. Mox-LDL treatment of HUVECs also altered the expression of proteins involved in hemostasis, cell adhesion, angiogenesis, inflammation and stress responses. In addition, proteins from the mitochondrial respiratory chain were upregulated after Mox-LDL treatment. Finally, a trihydroxy-unsaturated fatty acid was secreted by HUVECs exposed to Mox-LDLs and could serve as a biomarker of Mox-LDL exposure.</div></div><div><h3>Conclusions</h3><div>Our findings suggest that Mox-LDLs are internalized and degraded by HUVECs. They seem to induce increased mitochondrial activity and oxidative stress, likely mediated by reactive oxygen species. We believe that HUVECs activate cytoprotective antioxidant coping mechanisms (glutathione synthesis, heme oxygenase-1…) to survive. Mox-LDLs may also modulate hemostasis and inflammatory responses.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 12","pages":"Article 130865"},"PeriodicalIF":2.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145249494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Calcium oxalate alters lipid profiles and annexin A2 enrichment in membrane fractions of murine inner medullary collecting duct cells","authors":"Zenab Shahzad ,&nbsp;Ramish Rafay ,&nbsp;Saeed Khan ,&nbsp;Amir Kazory ,&nbsp;Nancy D. Denslow ,&nbsp;Abdel A. Alli","doi":"10.1016/j.bbagen.2025.130868","DOIUrl":"10.1016/j.bbagen.2025.130868","url":null,"abstract":"<div><div>The mechanism underlying calcium oxalate (CaOx) stone formation remains unclear, although recurrent kidney stone formation has been linked to injury of kidney tubular cells in response to their exposure to higher oxalate and CaOx crystals. We hypothesized that CaOx induces the externalization and membrane enrichment of the negatively charged phospholipid phosphatidylserine (PS), while simultaneously enhancing the expression of calcium-binding proteins as a protective mechanism against cell death in inner medullary collecting duct (IMCD) cells. Annexin A2, a calcium-dependent phospholipid-binding protein, may play a protective role in CaOx stone formation. Mouse IMCD (mIMCD3) cells were treated with CaOx before performing a targeted lipidomic analysis of membrane fractions. Additionally, mIMCD3 cells were challenged with methyl-beta-cyclodextrin (MBCD) PS liposomes, Annexin A2 was evaluated by Western blotting and densitometric analysis. Lipidomic analysis revealed an enrichment of multiple PS lipid species in membrane fractions of mIMCD3 cells treated with CaOx. Western blotting and densitometric analysis further demonstrated a significant increase in Annexin A2 protein expression in the membrane fractions of mIMCD3 cells treated with MBCD PS liposomes. CaOx and MBCD PS liposomes were found to differentially affect membrane fluidity in mIMCD3 cells. Importantly, CaOx treatment caused an increase in ACSL4 and caspase 9 protein expression, indicating initiation of ferroptosis and apoptosis, respectively. The siRNA mediated knockdown of annexin A2 further augmented the upregulation of caspase 9 after CaOx treatment. Collectively, these findings suggest that Annexin A2, a calcium-binding and plasma membrane repair protein, is upregulated in response to CaOx-induced PS enrichment in mIMCD3 cells.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1869 12","pages":"Article 130868"},"PeriodicalIF":2.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145285543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}