FEBS Open Bio最新文献

{"title":"Advanced glycation end products promote the release of endothelial cell-derived mitocytosis","authors":"Rong Liu,&nbsp;Yuhao Zhang,&nbsp;Tiantian Ge,&nbsp;Hao Wu,&nbsp;Yongbin Ma,&nbsp;Honghua Ye,&nbsp;Fei Guo","doi":"10.1002/2211-5463.70035","DOIUrl":"10.1002/2211-5463.70035","url":null,"abstract":"<p>Accumulation of advanced glycation end products (AGEs) and endothelial dysfunction are major factors that contribute to the progression of vascular complications in diabetes. Migrasomes, a newly discovered organelle involved in mitocytosis, play an important role in the selective removal of damaged mitochondria. Our research shows that human umbilical vein endothelial cells (HUVECs) can release migrasomes and undergo mitocytosis. In addition, when exposed to oxidative stress from AGEs, mitochondrial damage worsens, leading to the activation of migrasome-mediated mitocytosis. We also found that migrasomes carrying mitochondria can be taken up by recipient cells. Understanding the connection between migrasome release, mitocytosis, and mitochondrial function in endothelial cells sheds light on the biological processes behind intercellular communication.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 7","pages":"1068-1078"},"PeriodicalIF":2.8,"publicationDate":"2025-04-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70035","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143802894","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A non-fluorescent immunohistochemistry method for measuring autophagy flux using MAP1LC3/LC3 and SQSTM1 as core markers","authors":"Shahla Shojaei,&nbsp;Amir Barzegar Behrooz,&nbsp;Marco Cordani,&nbsp;Mahmoud Aghaei,&nbsp;Negar Azarpira,&nbsp;Daniel J. Klionsky,&nbsp;Saeid Ghavami","doi":"10.1002/2211-5463.70014","DOIUrl":"10.1002/2211-5463.70014","url":null,"abstract":"<p>Macroautophagy/autophagy is a crucial cellular process for degrading and recycling damaged proteins and organelles, playing a significant role in diseases such as cancer and neurodegeneration. Evaluating autophagy flux, which tracks autophagosome formation, maturation, and degradation, is essential for understanding disease mechanisms. Current fluorescence-based methods are resource-intensive, requiring advanced equipment and expertise, limiting their use in clinical laboratories. Here, we introduce a non-fluorescent immunohistochemistry (IHC) method using MAP1LC3/LC3 and SQSTM1 as core markers for autophagy flux assessment. LC3 levels reflect autophagosome formation, whereas SQSTM1 degradation and a decrease in the number of its puncta indicate active flux (i.e., lysosomal turnover). We optimized chromogenic detection using diaminobenzidine (DAB) staining and developed a scoring system based on puncta number and the percentage of stained cells. This accessible, cost-effective method enables reliable autophagy quantification using a standard light microscope, bridging the gap between experimental research and clinical diagnostics. Our protocol allows accurate autophagy evaluation in fixed tissues, offering practical applications in biomedical research and clinical pathology assessment.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 6","pages":"898-905"},"PeriodicalIF":2.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70014","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143779531","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adenosine A3 receptor antagonists as anti-tumor treatment in human prostate cancer: an in vitro study","authors":"Maria Beatrice Morelli,&nbsp;Andrea Spinaci,&nbsp;Cui Chang,&nbsp;Rosaria Volpini,&nbsp;Catia Lambertucci,&nbsp;Matteo Landriscina,&nbsp;Vincenza Conteduca,&nbsp;Consuelo Amantini,&nbsp;Cristina Aguzzi,&nbsp;Laura Zeppa,&nbsp;Martina Giangrossi,&nbsp;Laura Soverchia,&nbsp;Matteo Santoni,&nbsp;Massimo Nabissi,&nbsp;Giorgio Santoni,&nbsp;Carlo Polidori","doi":"10.1002/2211-5463.70024","DOIUrl":"10.1002/2211-5463.70024","url":null,"abstract":"<p>Prostate cancer (PCa) is one of the most common cancers in men, and for patients with PCa that cannot be surgically resected or treated, androgen suppression therapy often results in significant adverse effects. Recent studies have shown that A3 adenosine receptors (A₃ARs) are overexpressed in prostate cancer (PCa), and several A₃AR agonists and antagonists have been investigated as potential anticancer drugs. In this study, we investigated the potential therapeutic effects of the A₃AR antagonists AR 292 and AR 357 in human PCa cell lines. LNCaP, DU-145, and PC3 cell lines were treated with AR 292 and AR 357 compounds, and their cytotoxic effects were determined using viability assays, flow cytometry, and western blotting. Moreover, the drug transporter gene profile was evaluated using RT-PCR in untreated and A₃AR antagonist-treated PCa cells. Both AR 292 and AR 357 showed antiproliferative effects with significant cell cycle arrest and induced DNA damage leading to cell death. AR 292 and especially AR 357 modulated the expression of drug transporter genes involved in chemoresistance, ferroptosis, and the hypoxia response. Ferroptosis was induced in DU-145 cells treated with both compounds as well as in PC3 cells treated with AR 357. However, the treatment of PC3 cells with AR 292 and the treatment of LNCaP cells with both AR 292 and AR 357 resulted in necrotic cell death. In conclusion, our study showed that A₃AR ligands exert anticancer effects via different mechanisms on PCa cell lines through the activation of multiple molecular pathways.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 7","pages":"1159-1175"},"PeriodicalIF":2.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70024","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143779534","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correction to “HELQ deficiency impairs the induction of primordial germ cell-like cells”","authors":"","doi":"10.1002/2211-5463.70029","DOIUrl":"10.1002/2211-5463.70029","url":null,"abstract":"<p>Cong Wan, Yaping Huang, Xingguo Xue, Gang Chang, Mei Wang, Xiao-Yang Zhao, Fang Luo, Zhi-Zhong Tang. HELQ deficiency impairs the induction of primordial germ cell-like cells. <i>FEBS Open Bio</i>. 2024; 14(7): 1087–1100, https://doi.org/10.1002/2211-5463.13810</p><p>The original designation of first authorship was incorrect: Cong Wan was listed as the sole first author, while Yaping Huang was inadvertently omitted as a co-first author.</p><p>The authorship should have been amended to: Co-first authors: Cong Wan &amp; Yaping Huang.</p><p>We apologize for this error.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 5","pages":"885"},"PeriodicalIF":2.8,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70029","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143771628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improving experimental reproducibility through comprehensive research protocols","authors":"Ivana Novak","doi":"10.1002/2211-5463.70021","DOIUrl":"https://doi.org/10.1002/2211-5463.70021","url":null,"abstract":"<p>With this “In the Limelight: Research Protocols” special issue, <i>FEBS Open Bio</i> aims to highlight the critical importance of publishing reproducible and detailed scientific protocols that can be broadly adopted by laboratories working in molecular and cellular life sciences. This collection includes four protocols focused on sample preparation for structural analysis of macromolecules using X-ray crystallography, covering both <i>in vitro</i> and <i>in vivo</i> approaches. Two additional protocols demonstrate the use of cellular systems to screen the enzymatic activity of various proteins, with potential applications in high-throughput screening. Another protocol provides a comprehensive guide for the preparation and analysis of selective autophagy flux in cultured cells using flow cytometry. Finally, the issue concludes with a protocol integrating classical behavioral tests with cognitive components to assess both physical and cognitive dimensions of frailty.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 4","pages":"530-531"},"PeriodicalIF":2.8,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143749432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"On subcellular distribution of the zinc finger 469 protein (ZNF469) and observed discrepancy in the localization of endogenous and overexpressed ZNF469","authors":"Anne Elisabeth Christensen Mellgren,&nbsp;Ileana Cristea,&nbsp;Thomas Stevenson,&nbsp;Endy Spriet,&nbsp;Per Morten Knappskog,&nbsp;Stig Ove Bøe,&nbsp;Harald Kranz,&nbsp;Sushma N. Grellscheid,&nbsp;Eyvind Rødahl","doi":"10.1002/2211-5463.70034","DOIUrl":"10.1002/2211-5463.70034","url":null,"abstract":"<p>The zinc finger 469 gene (<i>ZNF469</i>) is a single-exon gene predicted to encode a protein of 3953 amino acids. Despite pathogenic <i>ZNF469</i> variants being associated with Brittle Cornea Syndrome (BCS), relatively little is known about ZNF469 beyond its participation in regulating the expression of genes encoding extracellular matrix proteins. In this study, we examined the expression and intracellular localization of ZNF469 in different cell lines. The level of ZNF469 mRNA varied from low levels in HEK293 cells to high levels in HeLa cells and primary fibroblasts. Antibodies against ZNF469 reacted among others with a protein of approximately 400 kDa in immunoblot analysis, which was mainly present in the insoluble fraction of the cytoplasm. Immunofluorescence analysis of interphase cells showed small cytoplasmic puncta and weak nuclear staining. In dividing HeLa cells, the antibodies recognized foci that also stained for proteasomes. In transfected cells, ZNF469 was observed mainly in foci resembling nuclear speckles in interphase and at the midbody during mitosis. The nuclear foci showed overlapping staining with proteasomes. In live cell imaging, liquid-like properties of the nuclear foci were recorded as they changed shape and position and occasionally fused with each other. During stress granule formation, cytoplasmic foci showed overlapping staining with G3BP1. Finally, <i>in silico</i> analysis revealed large intrinsically disordered regions with multiple low complexity domains in ZNF469. Our data indicate that ZNF469 forms aggregates possibly as biomolecular condensates when overexpressed. However, care must be taken when analyzing the intracellular distribution of ZNF469 due to the discrepancy in the localization of endogenous ZNF469 and overexpressed ZNF469 in transfected cells.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 7","pages":"1054-1067"},"PeriodicalIF":2.8,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70034","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143742613","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Intracellular protein crystallization in living insect cells","authors":"Robert Schönherr,&nbsp;Nina Eichler,&nbsp;Fatama A. Sornaly,&nbsp;Juliane Boger,&nbsp;Anne M. Frevert,&nbsp;Janine Mia Lahey-Rudolph,&nbsp;Hannah Meyer,&nbsp;Lisa Weymar,&nbsp;Lars Redecke","doi":"10.1002/2211-5463.70020","DOIUrl":"10.1002/2211-5463.70020","url":null,"abstract":"<p>Crystallization of recombinant proteins in living cells is an emerging approach complementing conventional crystallization techniques. Homogeneous microcrystals well suited for serial diffraction experiments at X-ray free-electron lasers and synchrotron sources can be produced in a quasi-native environment, without the need for target protein purification. Several protein structures have already been solved; however, exploiting the full potential of this approach requires a systematic and versatile screening strategy for intracellular crystal growth. Recently, we published InCellCryst, a streamlined pipeline for producing microcrystals within living insect cells. Here, we present the detailed protocol, including optimized target gene expression using a baculovirus vector system, crystal formation, detection, and serial X-ray diffraction directly in the cells. The specific environment within the different cellular compartments acts as a screening parameter to maximize the probability of crystal growth. If successful, diffraction data can be collected 24 days after the start of target gene cloning.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 4","pages":"551-562"},"PeriodicalIF":2.8,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70020","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143735660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Soman induces endoplasmic reticulum stress and apoptosis of cerebral organoids via the GRP78-ATF6-CHOP signaling pathway","authors":"Yue Wei,&nbsp;Zhanbiao Liu,&nbsp;Jingjing Shi,&nbsp;Qian Jin,&nbsp;Wenqian Chen,&nbsp;Xuejun Chen,&nbsp;Liqin Li,&nbsp;Hui Chen","doi":"10.1002/2211-5463.70027","DOIUrl":"10.1002/2211-5463.70027","url":null,"abstract":"<p>Soman is an organophosphorus compound that induces neurotoxicity. In addition to its direct toxic effects resulting from acetylcholine accumulation, neurotoxicity may also be exacerbated by inducing endoplasmic reticulum (ER) stress. In light of the current scarcity of appropriate <i>in vitro</i> assessment models, in the present study, we used cerebral organoids derived from human pluripotent stem cells, a new tool for investigating the mechanisms of neurotoxicity, to investigate soman-induced ER stress. The results demonstrated that soman significantly suppressed acetylcholinesterase activity and activated the GRP78-ATF6-CHOP (i.e. glucose-regulated protein 78-activating transcription factor 6-C/EBP homologous protein) ER stress cascade, driving apoptosis in cerebral organoids. Pharmacological inhibition of ER stress by pre-treating cerebral organoids with the ER stress inhibitor 4-phenylbutyric acid prior to soman exposure attenuated apoptotic signaling and downregulated GRP78, ATF6 and CHOP expression. Parallel <i>in vivo</i> validation utilized a rat model with subcutaneous soman exposure, focusing on hippocampal and striatal ER stress markers. Consistent with the <i>in vitro</i> findings, soman-exposed rats exhibited marked ER stress activation in brain regions critical for neurotoxicity. This study establishes ER stress as a key contributor to soman-induced neurotoxicity and highlights cerebral organoids as a physiologically relevant model for organophosphorus compound research. We propose ER stress modulation as a potential therapeutic strategy to mitigate neurotoxic outcomes.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 7","pages":"1041-1053"},"PeriodicalIF":2.8,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70027","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143729445","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Bioengineering facets of the tumor microenvironment in 3D tumor models: insights into cellular, biophysical and biochemical interactions.","authors":"Salma T Rafik, Deniz Bakkalci, Alexander J MacRobert, Umber Cheema","doi":"10.1002/2211-5463.70018","DOIUrl":"https://doi.org/10.1002/2211-5463.70018","url":null,"abstract":"<p><p>The hallmarks of cancer extend beyond genetic anomalies to encompass a sophisticated tumor microenvironment, involving interactions between cancer and non-cancer cells within a dynamic biophysical setting, influencing cancer progression. The tumor microenvironment is multifaceted, and it is increasingly clear that the interaction and interdependence of these different facets need to be better understood. Tissue engineering of 3D in vitro models of the tumor microenvironment provides an opportunity to study these interactions and their interdependence on cancer progression. Cancer metastasis still poses a major challenge, accounting for 90% of cancer-related deaths. This accentuates the critical need to establish patient-specific model systems that replicate tumor complexity at all stages of progression. Herein, we outline the latest advancements of in vitro 3D models of the tumor microenvironment and the different tools utilized to analyze such models. Henceforth, the interaction of the multifaceted tumor microenvironment can be elucidated using such sophisticated in vitro tools.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":" ","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143729440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Positive feedback loop between MAPK and aquaporin 7 regulates autophagy and apoptosis induced by palmitate in RIN-m5f cells","authors":"Maoqi Wang,&nbsp;Jiang Tan,&nbsp;Xueting He,&nbsp;Yuqin Chen,&nbsp;Guoping Qiu,&nbsp;Mei Yang","doi":"10.1002/2211-5463.70011","DOIUrl":"10.1002/2211-5463.70011","url":null,"abstract":"<p>Type 2 diabetes mellitus (T2DM) is characterized by peripheral blood insulin resistance and progressive pancreatic β-cell dysfunction which is closely related to apoptosis of β-cells. Aquaporin 7 (AQP7) is the only aquaglyceroporin protein expressed in pancreatic β-cells. However, the relationship between AQP7 and autophagy remains unexplored, with limited studies investigating its link to islet β-cell apoptosis. In our study, we utilized an <i>in vitro</i> model involving palmitate-treated rat pancreatic β-cells (RIN-m5f) to examine these relationships. Our aim was to investigate the effects of AQP7 on autophagy and apoptosis by examining LC3 lipidation levels and p62 expression in pancreatic islet β-cells, thereby elucidating potential underlying mechanisms. Our results showed that phosphorylation of p38 and c-Jun-terminal kinase (JNK) increased in response to palmitate treatment, indicating the activation of these signaling pathways. Conversely, AQP7 expression decreased, reduced autophagy, and promoted apoptosis. AQP7 knockdown activated the p38 and JNK signaling pathways, inhibited autophagy (as evidenced by LC3 lipidation and p62 expression), and increased apoptosis. Furthermore, AQP7 overexpression repressed palmitate-induced apoptosis and alleviated autophagy inhibition by suppressing the p38 and JNK mitogen-activated protein kinase (MAPK) signaling pathways. Our results suggest a positive feedback loop between MAPK signaling and AQP7 that regulates autophagy and apoptosis in RIN-m5f cells under high-fat conditions.</p>","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":"15 6","pages":"972-984"},"PeriodicalIF":2.8,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/2211-5463.70011","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143700110","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}