European journal of cell biology最新文献

{"title":"Phosphatidylserine on sperm head interact with Annexin A5 on oviduct luminal cilia to form a sperm reservoir in pigs.","authors":"Lorraine Schmaltz, Elie Barakat, Renaud Fleurot, Rustem Uzbekov, Karine Reynaud, Ludivine Laffont, Guillaume Tsikis, Isabelle Mérour, Pascal Mermillod, Marie Saint-Dizier","doi":"10.1016/j.ejcb.2024.151471","DOIUrl":"https://doi.org/10.1016/j.ejcb.2024.151471","url":null,"abstract":"<p><p>After insemination, a subpopulation of sperm reaches the oviducts and binds to isthmic epithelial cells to form a \"sperm reservoir\". Our objective was to explore the role of annexin A5 (ANXA5), a protein that binds with high affinity to phosphatidylserine (PS), in the formation of the sperm reservoir in pigs. Phosphatidylserine was detected on the head of approximately 10 % of boar sperm at ejaculation. Porcine ANXA5 was immunodetected with a strong signal on luminal cilia in the isthmus and in derived isthmic epithelial spheroids (IES). Exogenous PS between 0.01 and 0.1 µg/mL and recombinant porcine ANXA5 (rpANXA5) above 0.1 µg/mL inhibited sperm binding to IES without reducing sperm motility. Pre-incubation of sperm, but not IES, with rpANXA5 inhibited sperm binding to IES. Under capacitating conditions, the proportion of live sperm with head PS exposure and the ability of sperm to bind to rpANXA5 and IES cilia increased within 30 min. Conversely, the acrosome reaction decreased the ability of sperm to bind rpANXA5 and prevented sperm binding to IES. In conclusion, sperm membrane remodelling during capacitation enhanced head PS exposure in motile sperm, resulting in increased interaction with ciliary ANXA5 on isthmic epithelial spheroids. These findings support a role for PS-ANXA5 interaction in the formation of the sperm reservoir in mammalian females.</p>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"104 1","pages":"151471"},"PeriodicalIF":4.5,"publicationDate":"2024-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142863752","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":"GsMTx-4 venom toxin antagonizes biophysical modulation of metastatic traits in human osteosarcoma cells.","authors":"Arianna Buglione, Giulia Alloisio, Chiara Ciaccio, David Becerril Rodriguez, Simone Dogali, Marco Luce, Stefano Marini, Antonio Cricenti, Magda Gioia","doi":"10.1016/j.ejcb.2024.151469","DOIUrl":"https://doi.org/10.1016/j.ejcb.2024.151469","url":null,"abstract":"<p><p>Despite their genetic diversity, metastatic cells converge on similar physical constraints during tumor progression. At the nanoscale, these forces can induce substantial molecular deformations, altering the structure and behavior of cancer cells. To address the challenges of osteosarcoma (OS), a highly aggressive cancer, we explored the mechanobiology of OS cells, in vitro. Using uniaxial-stretching technology, we examined the biophysical modulation of metastatic traits in SAOS-2, U-2 OS, and non-tumorigenic hFOB cells. Changes in cell morphology were quantified using confocal and fluorescence microscopy. To elucidate the molecular mechanisms that translate biomechanical alterations into biochemical responses, we employed Western blotting, real-time quantitative RT-PCR, reactive oxygen species ROS assay, and the mechanosensitive channel blocker Grammostola MechanoToxin4 (GsMTx-4). Our study reveals that mechanical stimulation uniquely affects OS cells, increasing nuclear size and altering the N/C ratio. We found that mechanosensitive (MS) channels are activated, leading to ROS accumulation, Src protein modulation, and histone H3 acetylation. These changes influence OS cell motility and adhesion but not proliferation. Importantly, mechanical preconditioning differentially impacts doxorubicin resistance, correlating with the Src-H3 acetylation axis. This study underscores the critical role of MS channels in OS cells and highlights the importance of mechanobiology in identifying molecular pathways that traditional biochemical approaches may not reveal. Notably, the GsMTx-4 venom peptide effectively countered mechanically induced responses, particularly by inhibiting OS cell migration, without harming healthy cells. Thus, suggesting its potential as a promising therapeutic agent for targeting osteosarcoma metastasis.</p>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"104 1","pages":"151469"},"PeriodicalIF":4.5,"publicationDate":"2024-12-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142822017","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":"Editorial overview: EJCB Special Issue – Cell host-pathogen interactions","authors":"Serge Mostowy ,&nbsp;Theresia E.B. Stradal","doi":"10.1016/j.ejcb.2024.151462","DOIUrl":"10.1016/j.ejcb.2024.151462","url":null,"abstract":"","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 4","pages":"Article 151462"},"PeriodicalIF":4.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142497505","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Increased mTOR activity and RICTOR copy number in small cell lung carcinoma progression","authors":"Dániel Sztankovics ,&nbsp;Fatime Szalai ,&nbsp;Dorottya Moldvai ,&nbsp;Titanilla Dankó ,&nbsp;Noémi Nagy ,&nbsp;Judit Pápay ,&nbsp;András Khoór ,&nbsp;Ildikó Krencz ,&nbsp;Anna Sebestyén","doi":"10.1016/j.ejcb.2024.151468","DOIUrl":"10.1016/j.ejcb.2024.151468","url":null,"abstract":"<div><div>Small cell lung carcinoma (SCLC) is a highly malignant cancer with early metastatic dissemination and poor clinical outcomes. Mutations in the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway, including the frequently occurring rapamycin-insensitive protein (<em>RICTOR</em>) amplification, have been described in these tumours. Moreover, the associated mTOR hyperactivity could be exploited for personalised treatment. Our aim was to study mTOR activity, <em>RICTOR</em> amplification, and their role during SCLC progression. <em>In situ</em> mTOR activity and Rictor expression were characterised by immunohistochemistry in 50 primary and 50 brain metastatic tumours, and 14 paired SCLC patient samples. <em>RICTOR</em> copy number changes were analysed by fluorescence <em>in situ</em> hybridisation of the paired SCLC patient samples and <em>in vivo</em> experiments. Additionally, <em>in vitro</em> sensitivity to cisplatin and mTOR inhibitors was evaluated in SCLC cell lines harbouring <em>RICTOR</em> amplification and other mTOR pathway mutations. High Rictor expression and mTOR complex 2 (mTORC2) hyperactivity were significantly associated with brain metastases and worse overall survival. An increase in <em>RICTOR</em> copy number was observed in paired samples during progression. The importance of these alterations was confirmed both by the sensitising effect of vistusertib <em>in vitro</em> and the <em>RICTOR</em> copy number/expression changes in xenografts. Our study highlights the role of mTORC2 in SCLC progression. Early detection of <em>RICTOR</em> amplification in primary tumours and targeting mTORC2 in these cases may represent a promising novel strategy to develop personalised therapy for metastasis prevention.</div></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 4","pages":"Article 151468"},"PeriodicalIF":4.5,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142686320","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Gq/G11 oncogenic mutations promote PD-L1 expression and suppress tumor immunity","authors":"Jingyan Dong ,&nbsp;Yue Xu ,&nbsp;Dawei Yu ,&nbsp;Xiaoling Zhang ,&nbsp;Anqi Wang ,&nbsp;Lei Lv ,&nbsp;Zhiqing Li","doi":"10.1016/j.ejcb.2024.151467","DOIUrl":"10.1016/j.ejcb.2024.151467","url":null,"abstract":"<div><div>Uveal melanoma (UM) is the predominant form of eye cancer. The genes GNAQ and GNA11, encoding Gq and G11 respectively, are most frequently mutated in UM and are considered the major drivers of UM carcinogenesis by activating YAP. However, the mechanisms by which metastatic UM evades the immune system remain poorly understood. In this study, we found that oncogenic mutations of Gq/G11 promoted YAP and PD-L1 expression, modifying the tumor microenvironment and promoting immune evasion of UM. Consistently, the levels of GNAQ/GNA11 and YAP positively correlated to PD-L1 expression in UM patients. Furthermore, silencing YAP or treating with its inhibitor, Verteporfin, attenuated PD-L1 expression induced by Gq/G11 mutations, thereby enhancing T cell activation and T cell-mediated cytotoxicity. Collectively, this study reveals a potential role of Gq/G11 mutations on immune evasion of UM, a new mechanism of Gq/11 mutations-induced tumorigenesis, highlighting Gq/G11 and YAP as potential immunotherapeutic targets and suggesting Verteporfin as an adjuvant for immunotherapy of UM patients with GNAQ or GNA11 mutations.</div></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 4","pages":"Article 151467"},"PeriodicalIF":4.5,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142647186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation of the development and evolution of the mammalian cerebrum using gyrencephalic ferrets","authors":"Masanori Imamura ,&nbsp;Mayuko Yoshino ,&nbsp;Hiroshi Kawasaki","doi":"10.1016/j.ejcb.2024.151466","DOIUrl":"10.1016/j.ejcb.2024.151466","url":null,"abstract":"<div><div>Mammalian brains have evolved a neocortex, which has diverged in size and morphology in different species over the course of evolution. In some mammals, a substantial increase in the number of neurons and glial cells resulted in the expansion and folding of the cerebrum, and it is believed that these evolutionary changes contributed to the acquisition of higher cognitive abilities in mammals. However, their underlying molecular and cellular mechanisms remain insufficiently elucidated. A major difficulty in addressing these mechanisms stemmed from the lack of appropriate animal models, as conventional experimental animals such as mice and rats have small brains without structurally obvious folds. Therefore, researchers including us have focused on using ferrets instead of mice and rats. Ferrets are domesticated carnivorous mammals with a gyrencephalic cerebrum, and, notably, they are amenable to genetic manipulations including <em>in utero</em> electroporation to knock out genes in the cerebrum. In this review, we highlight recent research into the mechanisms underlying the development and evolution of cortical folds using ferrets.</div></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 4","pages":"Article 151466"},"PeriodicalIF":4.5,"publicationDate":"2024-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142638701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring optimal protocols for generating and preserving glucose-responsive insulin-secreting progenitor cells derived from human pluripotent stem cells","authors":"Patrizia Tornabene ,&nbsp;James M. Wells","doi":"10.1016/j.ejcb.2024.151464","DOIUrl":"10.1016/j.ejcb.2024.151464","url":null,"abstract":"<div><div>Human pluripotent stem cells (hPSCs) represent an unlimited source of β-like cells for both disease modeling and cellular therapy for diabetes. Numerous protocols have been published describing the differentiation of hPSCs into β-like cells that secret insulin in response to a glucose challenge. However, among the most widely used protocols it is not clear which yield the most functional cells with reproducible glucose-stimulated insulin-secretion (GSIS). Moreover, the technical challenges in culturing and differentiating hPSCs is a barrier for many researchers. In this study, we performed a side-by-side functional comparison based on three widely used methods to generate insulin expressing cells and identified optimal stages and conditions for cryopreserving and reconstituting stem cell (SC)-derived islets with a robust GSIS. Despite the fact that each protocol yields SC-islets consisting of insulin and glucagon-expressing cells, the SC-islets obtained from the two more recent revised protocols were more functional as measured by robust and reproducible GSIS. Moreover, we demonstrate that pancreatic progenitors and differentiated endocrine cells that have been cryopreserved for up to 10 months, can be reconstituted into glucose responsive SC-islets. These findings should enable the use of human PSC-derived β-like cells technologies even by groups with minimal PSC culture experience.</div></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 4","pages":"Article 151464"},"PeriodicalIF":4.5,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142561071","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An unexpected role of IL10 in mesoderm induction and differentiation from pluripotent stem cells: Implications in zebrafish angiogenic sprouting, vascular organoid development, and therapeutic angiogenesis","authors":"Kaiyuan Niu ,&nbsp;Chengxin Zhang ,&nbsp;Chenxin Liu ,&nbsp;Wei Wu ,&nbsp;Yi Yan ,&nbsp;Ancheng Zheng ,&nbsp;Silin Liu ,&nbsp;Zhenning Shi ,&nbsp;Mei Yang ,&nbsp;Wen Wang ,&nbsp;Qingzhong Xiao","doi":"10.1016/j.ejcb.2024.151465","DOIUrl":"10.1016/j.ejcb.2024.151465","url":null,"abstract":"<div><div>Mesoderm induction is a crucial step for vascular cell specification, vascular development and vasculogenesis. However, the cellular and molecular mechanisms underlying mesoderm induction remain elusive. In the present study, a chemically-defined differentiation protocol was used to induce mesoderm formation and generate functional vascular cells including smooth muscle cells (SMCs) and endothelial cells (ECs) from human induced pluripotent stem cells (hiPSCs). Zebrafish larvae were used to detect an in vivo function of interleukin 10 (IL10) in mesoderm formation and vascular development. A three dimensional approach was used to create hiPSC-derived blood vessel organoid (BVO) and explore a potential impact of IL10 on BVO formation. A murine model hind limb ischemia was applied to investigate a therapeutic potential of hiPSC-derived cells treated with or without IL10 during differentiation. We found that IL10 was significantly and specifically up-regulated during mesoderm stage of vascular differentiation. IL10 addition in mesoderm induction media dramatically increased mesoderm induction and vascular cell generation from hiPSCs, whereas an opposite effect was observed with IL10 inhibition. Mechanistic studies revealed that IL10 promotes mesoderm formation and vascular cell differentiation by activating signal transducer and activator of transcription 3 signal pathway. Functional studies with an <em>in vivo</em> model system confirmed that knockdown of IL10 using morpholino antisense oligonucleotides in zebrafish larvae caused defective mesoderm formation, angiogenic sprouting and vascular development. Additionally, our data also show IL10 promotes blood vessel organoid development and enhances vasculogenesis and angiogenesis. Importantly, we demonstrate that IL10 treatment during mesoderm induction stage enhances blood flow perfusion recovery and increases vasculogenesis and therapeutic angiogenesis after hind limb ischemia. Our data, therefore, demonstrate a regulatory role for IL10 in mesoderm formation from hiPSCs and during zebrafish vascular development, providing novel insights into mesoderm induction and vascular cell specifications.</div></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 4","pages":"Article 151465"},"PeriodicalIF":4.5,"publicationDate":"2024-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142544577","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"dCas9-HDAC8-EGFP fusion enables epigenetic editing of breast cancer cells by H3K9 deacetylation","authors":"Mohammad Mijanur Rahman ,&nbsp;Trygve O. Tollefsbol","doi":"10.1016/j.ejcb.2024.151463","DOIUrl":"10.1016/j.ejcb.2024.151463","url":null,"abstract":"<div><div>Epigenetic editing is thriving as a robust tool for manipulating transcriptional regulation and cell fate. Despite its regulatory role in gene downregulation, epigenetic editing with histone deacetylation has been sparsely studied, especially in the context of cancer. In this current study, we have reconstructed a dCas9-HDAC8-EGFP fusion to perform histone deacetylation on the promoter of the <em>ESR1</em>, <em>TERT</em> and <em>CDKN1C</em> genes for the first time in breast cancer cell lines MCF-7 and MDA-MB-231 as well as in HEK293T cells. Our results demonstrated that dCas9-HDAC8-EGFP in combination with appropriate gRNAs were able to downregulate the expression of the <em>ESR1</em>, <em>TERT</em> and <em>CDKN1C</em> genes transcriptionally by specifically depleting the H3K9ac level on the recruitment loci. The addition of histone deacetylase inhibitors was found to neutralize the outcomes of dCas9-HDAC8-EGFP-induced epigenetic editing. Furthermore, we observed a significant downregulation of full length ERα expression in epigenetically edited MCF-7 cells with consequential alteration in cellular response toward estradiol and tamoxifen treatment due to dCas9-HDAC8-EGFP mediated epigenetic editing of the <em>ESR1</em> gene. Overall, dCas9-HDAC8-EGFP is a novel circuit that enabled downregulation of crucial genes with cellular outcome in breast cancer cells by preferentially inducing H3K9 deacetylation of specific promoter regions.</div></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 4","pages":"Article 151463"},"PeriodicalIF":4.5,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142497504","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The double deficiency of the SNARE proteins vti1a and vti1b affects neurite outgrowth and signaling in N1E-115 neuroblastoma cells.","authors":"Katharina Kotschnew ,&nbsp;Denise Winkler ,&nbsp;Jonas Reckmann ,&nbsp;Charlotte Mann ,&nbsp;Alina Schweigert ,&nbsp;Greta Tellkamp ,&nbsp;Kristian M. Müller ,&nbsp;Gabriele Fischer von Mollard","doi":"10.1016/j.ejcb.2024.151461","DOIUrl":"10.1016/j.ejcb.2024.151461","url":null,"abstract":"<div><div>During intracellular trafficking N-ethylmaleimide-sensitive-factor attachment receptor (SNARE) proteins catalyze the membrane fusion by assembling into a four-helix complex. In the mouse model, loss of the endosomal SNAREs vti1a and vti1b results in a perinatal lethal phenotype and neuronal defects including decreased neurite outgrowth in cultured primary neurons.</div><div>We used a CRISPR/Cas9 system to generate a <em>Vti1a Vti1b</em> double knockout (DKO) in the neuroblastoma cell line N1E-115. Three different DKO cell lines were obtained and examined at genome and protein level. The double deficiency impaired proper differentiation based on lower levels of synaptic proteins as well as reduced neurite formation and elongation compared to wild type cells in differentiation medium. Neurite elongation can be induced by a variety of extracellular signals via different signaling cascades. Treatment with the Rho kinase inhibitor Y27632, which stimulates enlargeosome exocytosis, or with neurotrophic factors (BDNF, NGF and NT3) resulted in reduced stimulation of all DKO clones and in significantly shorter neurites compared to wild type cells. The loss of vti1a and vti1b disrupted Akt signaling during enlargeosome-mediated and Erk signaling during BDNF-induced neurite outgrowth.</div></div>","PeriodicalId":12010,"journal":{"name":"European journal of cell biology","volume":"103 4","pages":"Article 151461"},"PeriodicalIF":4.5,"publicationDate":"2024-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142425283","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}