Inhibition of exosome biogenesis affects cell motility in heterogeneous sub-populations of paediatric-type diffuse high-grade gliomas
Abstract Background Paediatric-type diffuse High-Grade Gliomas (PDHGG) are highly heterogeneous tumours which include distinct cell sub-populations co-existing within the same tumour mass. We have previously shown that primary patient-derived and optical barcoded single-cell-derived clones function...
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| Format: | Article |
| Language: | English |
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BMC
2023-11-01
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| Series: | Cell & Bioscience |
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| Online Access: | https://doi.org/10.1186/s13578-023-01166-5 |
| _version_ | 1797556190966710272 |
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| author | Giulia Pericoli Angela Galardi Alessandro Paolini Lucia Lisa Petrilli Gerardo Pepe Alessandro Palma Marta Colletti Roberta Ferretti Ezio Giorda Stefano Levi Mortera Anna Burford Andrea Carai Angela Mastronuzzi Alan Mackay Lorenza Putignani Chris Jones Luisa Pascucci Hector Peinado Manuela Helmer-Citterich Emmanuel de Billy Andrea Masotti Franco Locatelli Angela Di Giannatale Maria Vinci |
| author_facet | Giulia Pericoli Angela Galardi Alessandro Paolini Lucia Lisa Petrilli Gerardo Pepe Alessandro Palma Marta Colletti Roberta Ferretti Ezio Giorda Stefano Levi Mortera Anna Burford Andrea Carai Angela Mastronuzzi Alan Mackay Lorenza Putignani Chris Jones Luisa Pascucci Hector Peinado Manuela Helmer-Citterich Emmanuel de Billy Andrea Masotti Franco Locatelli Angela Di Giannatale Maria Vinci |
| author_sort | Giulia Pericoli |
| collection | DOAJ |
| description | Abstract Background Paediatric-type diffuse High-Grade Gliomas (PDHGG) are highly heterogeneous tumours which include distinct cell sub-populations co-existing within the same tumour mass. We have previously shown that primary patient-derived and optical barcoded single-cell-derived clones function as interconnected networks. Here, we investigated the role of exosomes as a route for inter-clonal communication mediating PDHGG migration and invasion. Results A comprehensive characterisation of seven optical barcoded single-cell-derived clones obtained from two patient-derived cell lines was performed. These analyses highlighted extensive intra-tumour heterogeneity in terms of genetic and transcriptional profiles between clones as well as marked phenotypic differences including distinctive motility patterns. Live single-cell tracking analysis of 3D migration and invasion assays showed that the single-cell-derived clones display a higher speed and longer travelled distance when in co-culture compared to mono-culture conditions. To determine the role of exosomes in PDHGG inter-clonal cross-talks, we isolated exosomes released by different clones and characterised them in terms of marker expression, size and concentration. We demonstrated that exosomes are actively internalized by the cells and that the inhibition of their biogenesis, using the phospholipase inhibitor GW4689, significantly reduced the cell motility in mono-culture and more prominently when the cells from the clones were in co-culture. Analysis of the exosomal miRNAs, performed with a miRNome PCR panel, identified clone-specific miRNAs and a set of miRNA target genes involved in the regulation of cell motility/invasion/migration. These genes were found differentially expressed in co-culture versus mono-culture conditions and their expression levels were significantly modulated upon inhibition of exosome biogenesis. Conclusions In conclusion, our study highlights for the first time a key role for exosomes in the inter-clonal communication in PDHGG and suggests that interfering with the exosome biogenesis pathway may be a valuable strategy to inhibit cell motility and dissemination for these specific diseases. |
| first_indexed | 2024-03-10T16:58:23Z |
| format | Article |
| id | doaj.art-d7785bfb848a4134a77dbd73c0fde68b |
| institution | Directory Open Access Journal |
| issn | 2045-3701 |
| language | English |
| last_indexed | 2024-03-10T16:58:23Z |
| publishDate | 2023-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell & Bioscience |
| spelling | doaj.art-d7785bfb848a4134a77dbd73c0fde68b2023-11-20T11:03:56ZengBMCCell & Bioscience2045-37012023-11-0113112410.1186/s13578-023-01166-5Inhibition of exosome biogenesis affects cell motility in heterogeneous sub-populations of paediatric-type diffuse high-grade gliomasGiulia Pericoli0Angela Galardi1Alessandro Paolini2Lucia Lisa Petrilli3Gerardo Pepe4Alessandro Palma5Marta Colletti6Roberta Ferretti7Ezio Giorda8Stefano Levi Mortera9Anna Burford10Andrea Carai11Angela Mastronuzzi12Alan Mackay13Lorenza Putignani14Chris Jones15Luisa Pascucci16Hector Peinado17Manuela Helmer-Citterich18Emmanuel de Billy19Andrea Masotti20Franco Locatelli21Angela Di Giannatale22Maria Vinci23Department of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSDepartment of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSMultifactorial and Complex Phenotype Research Area, Bambino Gesù Children’s Hospital-IRCCSDepartment of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSDepartment of Biology, University of Rome “Tor Vergata”Department of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSDepartment of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSDepartment of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSCore Facilities research laboratories, Bambino Gesù Children’s Hospital-IRCCSMultimodal Laboratory Medicine Research Area, Bambino Gesù Children’s Hospital, IRCCSDepartment of Molecular Pathology, The Institute of Cancer ResearchOncological Neurosurgery Unit, Department of Neuroscience and Neurorehabilitation, Bambino Gesù Children’s Hospital-IRCCSDepartment of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSDepartment of Molecular Pathology, The Institute of Cancer ResearchMultimodal Laboratory Medicine Research Area, Bambino Gesù Children’s Hospital, IRCCSDepartment of Molecular Pathology, The Institute of Cancer ResearchDepartment of Veterinary Medicine, University of PerugiaMicroenvironment & Metastasis Group, Molecular Oncology Program, Spanish National Cancer Research Centre (CNIO)Department of Biology, University of Rome “Tor Vergata”Department of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSMultifactorial and Complex Phenotype Research Area, Bambino Gesù Children’s Hospital-IRCCSDepartment of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSDepartment of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSDepartment of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children’s Hospital-IRCCSAbstract Background Paediatric-type diffuse High-Grade Gliomas (PDHGG) are highly heterogeneous tumours which include distinct cell sub-populations co-existing within the same tumour mass. We have previously shown that primary patient-derived and optical barcoded single-cell-derived clones function as interconnected networks. Here, we investigated the role of exosomes as a route for inter-clonal communication mediating PDHGG migration and invasion. Results A comprehensive characterisation of seven optical barcoded single-cell-derived clones obtained from two patient-derived cell lines was performed. These analyses highlighted extensive intra-tumour heterogeneity in terms of genetic and transcriptional profiles between clones as well as marked phenotypic differences including distinctive motility patterns. Live single-cell tracking analysis of 3D migration and invasion assays showed that the single-cell-derived clones display a higher speed and longer travelled distance when in co-culture compared to mono-culture conditions. To determine the role of exosomes in PDHGG inter-clonal cross-talks, we isolated exosomes released by different clones and characterised them in terms of marker expression, size and concentration. We demonstrated that exosomes are actively internalized by the cells and that the inhibition of their biogenesis, using the phospholipase inhibitor GW4689, significantly reduced the cell motility in mono-culture and more prominently when the cells from the clones were in co-culture. Analysis of the exosomal miRNAs, performed with a miRNome PCR panel, identified clone-specific miRNAs and a set of miRNA target genes involved in the regulation of cell motility/invasion/migration. These genes were found differentially expressed in co-culture versus mono-culture conditions and their expression levels were significantly modulated upon inhibition of exosome biogenesis. Conclusions In conclusion, our study highlights for the first time a key role for exosomes in the inter-clonal communication in PDHGG and suggests that interfering with the exosome biogenesis pathway may be a valuable strategy to inhibit cell motility and dissemination for these specific diseases.https://doi.org/10.1186/s13578-023-01166-5Paediatric-type diffuse high-grade gliomaDIPGGBMHeterogeneityCell communicationExosome |
| spellingShingle | Giulia Pericoli Angela Galardi Alessandro Paolini Lucia Lisa Petrilli Gerardo Pepe Alessandro Palma Marta Colletti Roberta Ferretti Ezio Giorda Stefano Levi Mortera Anna Burford Andrea Carai Angela Mastronuzzi Alan Mackay Lorenza Putignani Chris Jones Luisa Pascucci Hector Peinado Manuela Helmer-Citterich Emmanuel de Billy Andrea Masotti Franco Locatelli Angela Di Giannatale Maria Vinci Inhibition of exosome biogenesis affects cell motility in heterogeneous sub-populations of paediatric-type diffuse high-grade gliomas Cell & Bioscience Paediatric-type diffuse high-grade glioma DIPG GBM Heterogeneity Cell communication Exosome |
| title | Inhibition of exosome biogenesis affects cell motility in heterogeneous sub-populations of paediatric-type diffuse high-grade gliomas |
| title_full | Inhibition of exosome biogenesis affects cell motility in heterogeneous sub-populations of paediatric-type diffuse high-grade gliomas |
| title_fullStr | Inhibition of exosome biogenesis affects cell motility in heterogeneous sub-populations of paediatric-type diffuse high-grade gliomas |
| title_full_unstemmed | Inhibition of exosome biogenesis affects cell motility in heterogeneous sub-populations of paediatric-type diffuse high-grade gliomas |
| title_short | Inhibition of exosome biogenesis affects cell motility in heterogeneous sub-populations of paediatric-type diffuse high-grade gliomas |
| title_sort | inhibition of exosome biogenesis affects cell motility in heterogeneous sub populations of paediatric type diffuse high grade gliomas |
| topic | Paediatric-type diffuse high-grade glioma DIPG GBM Heterogeneity Cell communication Exosome |
| url | https://doi.org/10.1186/s13578-023-01166-5 |
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