FEBS Open Bio最新文献

Co-expression of HSV-1 ICP34.5 enhances the expression of gene delivered by self-amplifying RNA and mitigates its immunogenicity. HSV-1 ICP34.5的共表达增强了自扩增RNA传递基因的表达，降低了其免疫原性。

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-04-09 DOI: 10.1002/2211-5463.70036

Xuemin Lu, Yabin Wu, Chunye Zhao, Jie Zheng, Shangwu Chen, Yigang Wang, Yulong Xia

{"title":"Co-expression of HSV-1 ICP34.5 enhances the expression of gene delivered by self-amplifying RNA and mitigates its immunogenicity.","authors":"Xuemin Lu, Yabin Wu, Chunye Zhao, Jie Zheng, Shangwu Chen, Yigang Wang, Yulong Xia","doi":"10.1002/2211-5463.70036","DOIUrl":"https://doi.org/10.1002/2211-5463.70036","url":null,"abstract":"Self-amplifying RNA (saRNA) vectors have garnered significant attention for their potential in transient recombinant protein expression and vaccination strategies. These vectors are notable for their safety and the ability to produce high levels of protein from minimal input templates, offering a promising avenue for gene therapy applications. Despite their advantages, saRNA vectors face a critical challenge in their propensity to trigger a robust innate immune response. The presence of double-stranded RNA intermediates during saRNA replication activates pattern recognition receptors (PRRs), leading to the activation of protein kinase R (PKR) and interferon (IFN) signaling, which can result in a general translational shutdown within the host cell. To mitigate the stimulatory effects on PRRs and enhance the translation efficiency of saRNA, this study employs the saRNA-encoding HSV-1 neurovirulence protein ICP34.5, which is known for its ability to counteract the effects of PKR activation, potentially improving the translation efficiency of saRNA. It was shown that saRNA-encoding ICP34.5 clearly mediated the eukaryotic initiation factor 2 alpha subunit (eIF2α) dephosphorylation and significant suppression of innate immune responses in vitro, leading to enhanced expression of saRNA-encoded genes. The application of ICP34.5 incorporating saRNA vectors offers a more efficient and cost-effective solution for the production of proteins and the development of vaccines. This strategy could revolutionize the fields where saRNA utilization is envisioned, particularly in neurotropic disease applications where HSV-1 proteins may offer additional benefits.","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":" ","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143810871","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}

引用次数: 0

Advanced glycation end products promote the release of endothelial cell-derived mitocytosis. 晚期糖基化终产物促进内皮细胞衍生的有丝分裂的释放。

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-04-08 DOI: 10.1002/2211-5463.70035

Rong Liu, Yuhao Zhang, Tiantian Ge, Hao Wu, Yongbin Ma, Honghua Ye, Fei Guo

引用次数: 0

A non-fluorescent immunohistochemistry method for measuring autophagy flux using MAP1LC3/LC3 and SQSTM1 as core markers. 以MAP1LC3/LC3和SQSTM1为核心标记物的非荧光免疫组织化学方法测定自噬通量。

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-04-03 DOI: 10.1002/2211-5463.70014

Shahla Shojaei, Amir Barzegar Behrooz, Marco Cordani, Mahmoud Aghaei, Negar Azarpira, Daniel J Klionsky, Saeid Ghavami

{"title":"A non-fluorescent immunohistochemistry method for measuring autophagy flux using MAP1LC3/LC3 and SQSTM1 as core markers.","authors":"Shahla Shojaei, Amir Barzegar Behrooz, Marco Cordani, Mahmoud Aghaei, Negar Azarpira, Daniel J Klionsky, Saeid Ghavami","doi":"10.1002/2211-5463.70014","DOIUrl":"https://doi.org/10.1002/2211-5463.70014","url":null,"abstract":"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.","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":" ","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","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":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Adenosine A₃ receptor antagonists as anti-tumor treatment in human prostate cancer: an in vitro study. 腺苷A3受体拮抗剂在人前列腺癌中的抗肿瘤治疗作用：体外研究。

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-04-03 DOI: 10.1002/2211-5463.70024

Maria Beatrice Morelli, Andrea Spinaci, Cui Chang, Rosaria Volpini, Catia Lambertucci, Matteo Landriscina, Vincenza Conteduca, Consuelo Amantini, Cristina Aguzzi, Laura Zeppa, Martina Giangrossi, Laura Soverchia, Matteo Santoni, Massimo Nabissi, Giorgio Santoni, Carlo Polidori

{"title":"Adenosine A3 receptor antagonists as anti-tumor treatment in human prostate cancer: an in vitro study.","authors":"Maria Beatrice Morelli, Andrea Spinaci, Cui Chang, Rosaria Volpini, Catia Lambertucci, Matteo Landriscina, Vincenza Conteduca, Consuelo Amantini, Cristina Aguzzi, Laura Zeppa, Martina Giangrossi, Laura Soverchia, Matteo Santoni, Massimo Nabissi, Giorgio Santoni, Carlo Polidori","doi":"10.1002/2211-5463.70024","DOIUrl":"https://doi.org/10.1002/2211-5463.70024","url":null,"abstract":"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.","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":" ","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-04-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","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":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Correction to "HELQ deficiency impairs the induction of primordial germ cell-like cells". 更正“HELQ缺乏损害原始生殖细胞样细胞的诱导”。

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-04-02 DOI: 10.1002/2211-5463.70029

引用次数: 0

Improving experimental reproducibility through comprehensive research protocols 通过全面的研究方案提高实验的可重复性

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-04-01 DOI: 10.1002/2211-5463.70021

Ivana Novak

引用次数: 0

On subcellular distribution of the zinc finger 469 protein (ZNF469) and observed discrepancy in the localization of endogenous and overexpressed ZNF469. 锌指469蛋白（ZNF469）的亚细胞分布及内源性和过表达ZNF469的定位差异

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-03-29 DOI: 10.1002/2211-5463.70034

Anne Elisabeth Christensen Mellgren, Ileana Cristea, Thomas Stevenson, Endy Spriet, Per Morten Knappskog, Stig Ove Bøe, Harald Kranz, Sushma N Grellscheid, Eyvind Rødahl

{"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, Ileana Cristea, Thomas Stevenson, Endy Spriet, Per Morten Knappskog, Stig Ove Bøe, Harald Kranz, Sushma N Grellscheid, Eyvind Rødahl","doi":"10.1002/2211-5463.70034","DOIUrl":"https://doi.org/10.1002/2211-5463.70034","url":null,"abstract":"The zinc finger 469 gene (ZNF469) is a single-exon gene predicted to encode a protein of 3953 amino acids. Despite pathogenic ZNF469 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, in silico 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.","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":" ","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","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":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Intracellular protein crystallization in living insect cells 活昆虫细胞内蛋白质结晶。

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-03-28 DOI: 10.1002/2211-5463.70020

Robert Schönherr, Nina Eichler, Fatama A. Sornaly, Juliane Boger, Anne M. Frevert, Janine Mia Lahey-Rudolph, Hannah Meyer, Lisa Weymar, Lars Redecke

{"title":"Intracellular protein crystallization in living insect cells","authors":"Robert Schönherr, Nina Eichler, Fatama A. Sornaly, Juliane Boger, Anne M. Frevert, Janine Mia Lahey-Rudolph, Hannah Meyer, Lisa Weymar, Lars Redecke","doi":"10.1002/2211-5463.70020","DOIUrl":"10.1002/2211-5463.70020","url":null,"abstract":"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.","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}

引用次数: 0

Soman induces endoplasmic reticulum stress and apoptosis of cerebral organoids via the GRP78-ATF6-CHOP signaling pathway. Soman通过GRP78-ATF6-CHOP信号通路诱导脑类器官内质网应激和凋亡。

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-03-28 DOI: 10.1002/2211-5463.70027

Yue Wei, Zhanbiao Liu, Jingjing Shi, Qian Jin, Wenqian Chen, Xuejun Chen, Liqin Li, Hui Chen

{"title":"Soman induces endoplasmic reticulum stress and apoptosis of cerebral organoids via the GRP78-ATF6-CHOP signaling pathway.","authors":"Yue Wei, Zhanbiao Liu, Jingjing Shi, Qian Jin, Wenqian Chen, Xuejun Chen, Liqin Li, Hui Chen","doi":"10.1002/2211-5463.70027","DOIUrl":"https://doi.org/10.1002/2211-5463.70027","url":null,"abstract":"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 in vitro 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 in vivo validation utilized a rat model with subcutaneous soman exposure, focusing on hippocampal and striatal ER stress markers. Consistent with the in vitro 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.","PeriodicalId":12187,"journal":{"name":"FEBS Open Bio","volume":" ","pages":""},"PeriodicalIF":2.8,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","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":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Bioengineering facets of the tumor microenvironment in 3D tumor models: insights into cellular, biophysical and biochemical interactions. 三维肿瘤模型中肿瘤微环境的生物工程方面：对细胞，生物物理和生化相互作用的见解。

IF 2.8 4区生物学

FEBS Open Bio Pub Date : 2025-03-27 DOI: 10.1002/2211-5463.70018

Salma T Rafik, Deniz Bakkalci, Alexander J MacRobert, Umber Cheema

{"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":"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.","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}

引用次数: 0