Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cells
Abstract Background It has been proposed that mesenchymal stromal cells (MSCs) promote tumor progression by interacting with tumor cells and other stroma cells in the complex network of the tumor microenvironment. We characterized MSCs isolated and expanded from tumor tissues of pediatric patients d...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-11-01
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| Series: | BMC Cancer |
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| Online Access: | http://link.springer.com/article/10.1186/s12885-018-5082-2 |
| _version_ | 1831683753171222528 |
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| author | Gloria Pelizzo Veronica Veschi Melissa Mantelli Stefania Croce Vincenzo Di Benedetto Paolo D’Angelo Alice Maltese Laura Catenacci Tiziana Apuzzo Emanuela Scavo Antonia Moretta Matilde Todaro Giorgio Stassi Maria Antonietta Avanzini Valeria Calcaterra |
| author_facet | Gloria Pelizzo Veronica Veschi Melissa Mantelli Stefania Croce Vincenzo Di Benedetto Paolo D’Angelo Alice Maltese Laura Catenacci Tiziana Apuzzo Emanuela Scavo Antonia Moretta Matilde Todaro Giorgio Stassi Maria Antonietta Avanzini Valeria Calcaterra |
| author_sort | Gloria Pelizzo |
| collection | DOAJ |
| description | Abstract Background It has been proposed that mesenchymal stromal cells (MSCs) promote tumor progression by interacting with tumor cells and other stroma cells in the complex network of the tumor microenvironment. We characterized MSCs isolated and expanded from tumor tissues of pediatric patients diagnosed with neuroblastomas (NB-MSCs) to define interactions with the tumor microenvironment. Methods Specimens were obtained from 7 pediatric patients diagnosed with neuroblastoma (NB). Morphology, immunophenotype, differentiation capacity, proliferative growth, expression of stemness and neural differentiation markers were evaluated. Moreover, the ability of cells to modulate the immune response, i.e. inhibition of phytohemagglutinin (PHA) activated peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cytotoxic function, was examined. Gene expression profiles, known to be related to tumor cell stemness, Wnt pathway activation, epithelial-mesenchymal transition (EMT) and tumor metastasis were also evaluated. Healthy donor bone marrow-derived MSCs (BM-MSC) were employed as controls. Results NB-MSCs presented the typical MSC morphology and phenotype. They showed a proliferative capacity superimposable to BM-MSCs. Stemness marker expression (Sox2, Nanog, Oct3/4) was comparable to BM-MSCs. NB-MSC in vitro osteogenic and chondrogenic differentiation was similar to BM-MSCs, but NB-MSCs lacked adipogenic differentiation capacity. NB-MSCs reached senescence phases at a median passage of P7 (range, P5-P13). NB-MSCs exhibited greater immunosuppressive capacity on activated T lymphocytes at a 1:2 (MSC: PBMC) ratio compared with BM-MSCs (p = 0.018). NK cytotoxic activity was not influenced by co-culture, either with BM-MSCs or NB-MSCs. Flow-cytometry cell cycle analysis showed that NB-MSCs had an increased number of cells in the G0-G1 phase compared to BM-MSCs. Transcriptomic profiling results indicated that NB-MSCs were enriched with EMT genes compared to BM-MSCs. Conclusions We characterized the biological features, the immunomodulatory capacity and the gene expression profile of NB-MSCs. The NB-MSC gene expression profile and their functional properties suggest a potential role in promoting tumor escape, invasiveness and metastatic traits of NB cancer cells. A better understanding of the complex mechanisms underlying the interactions between NB cells and NB-derived MSCs should shed new light on potential novel therapeutic approaches. |
| first_indexed | 2024-12-20T06:44:34Z |
| format | Article |
| id | doaj.art-cd655a35087247ba8ddcbb0d33749385 |
| institution | Directory Open Access Journal |
| issn | 1471-2407 |
| language | English |
| last_indexed | 2024-12-20T06:44:34Z |
| publishDate | 2018-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Cancer |
| spelling | doaj.art-cd655a35087247ba8ddcbb0d337493852022-12-21T19:49:47ZengBMCBMC Cancer1471-24072018-11-0118111210.1186/s12885-018-5082-2Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cellsGloria Pelizzo0Veronica Veschi1Melissa Mantelli2Stefania Croce3Vincenzo Di Benedetto4Paolo D’Angelo5Alice Maltese6Laura Catenacci7Tiziana Apuzzo8Emanuela Scavo9Antonia Moretta10Matilde Todaro11Giorgio Stassi12Maria Antonietta Avanzini13Valeria Calcaterra14Pediatric Surgery Department, Children’s Hospital G. Di Cristina, ARNAS Civico-Di Cristina-BenfratelliCellular and Molecular Pathophysiology Laboratory, Department of Surgical, Oncological and Stomatological Sciences, University of PalermoImmunology and Transplantation Laboratory, Cell Factory, Pediatric Hematology Oncology Unit, Department of Maternal and Children’s Health, Fondazione IRCCS Policlinico S. MatteoImmunology and Transplantation Laboratory, Cell Factory, Pediatric Hematology Oncology Unit, Department of Maternal and Children’s Health, Fondazione IRCCS Policlinico S. MatteoPediatric Surgery Unit and NICU Policlinico-Vittorio Emanuele HospitalPediatric Hematology Oncology Unit, Children’s Hospital G. Di Cristina, ARNAS Civico-Di Cristina-BenfratelliImmunology and Transplantation Laboratory, Cell Factory, Pediatric Hematology Oncology Unit, Department of Maternal and Children’s Health, Fondazione IRCCS Policlinico S. MatteoImmunology and Transplantation Laboratory, Cell Factory, Pediatric Hematology Oncology Unit, Department of Maternal and Children’s Health, Fondazione IRCCS Policlinico S. MatteoCellular and Molecular Pathophysiology Laboratory, Department of Surgical, Oncological and Stomatological Sciences, University of PalermoCellular and Molecular Pathophysiology Laboratory, Department of Surgical, Oncological and Stomatological Sciences, University of PalermoImmunology and Transplantation Laboratory, Cell Factory, Pediatric Hematology Oncology Unit, Department of Maternal and Children’s Health, Fondazione IRCCS Policlinico S. MatteoDepartment of DIBIMIS, University of PalermoCellular and Molecular Pathophysiology Laboratory, Department of Surgical, Oncological and Stomatological Sciences, University of PalermoImmunology and Transplantation Laboratory, Cell Factory, Pediatric Hematology Oncology Unit, Department of Maternal and Children’s Health, Fondazione IRCCS Policlinico S. MatteoPediatrics and Adolescentology Unit, Department of Internal Medicine, University of Pavia, Fondazione IRCCS Policlinico San MatteoAbstract Background It has been proposed that mesenchymal stromal cells (MSCs) promote tumor progression by interacting with tumor cells and other stroma cells in the complex network of the tumor microenvironment. We characterized MSCs isolated and expanded from tumor tissues of pediatric patients diagnosed with neuroblastomas (NB-MSCs) to define interactions with the tumor microenvironment. Methods Specimens were obtained from 7 pediatric patients diagnosed with neuroblastoma (NB). Morphology, immunophenotype, differentiation capacity, proliferative growth, expression of stemness and neural differentiation markers were evaluated. Moreover, the ability of cells to modulate the immune response, i.e. inhibition of phytohemagglutinin (PHA) activated peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cytotoxic function, was examined. Gene expression profiles, known to be related to tumor cell stemness, Wnt pathway activation, epithelial-mesenchymal transition (EMT) and tumor metastasis were also evaluated. Healthy donor bone marrow-derived MSCs (BM-MSC) were employed as controls. Results NB-MSCs presented the typical MSC morphology and phenotype. They showed a proliferative capacity superimposable to BM-MSCs. Stemness marker expression (Sox2, Nanog, Oct3/4) was comparable to BM-MSCs. NB-MSC in vitro osteogenic and chondrogenic differentiation was similar to BM-MSCs, but NB-MSCs lacked adipogenic differentiation capacity. NB-MSCs reached senescence phases at a median passage of P7 (range, P5-P13). NB-MSCs exhibited greater immunosuppressive capacity on activated T lymphocytes at a 1:2 (MSC: PBMC) ratio compared with BM-MSCs (p = 0.018). NK cytotoxic activity was not influenced by co-culture, either with BM-MSCs or NB-MSCs. Flow-cytometry cell cycle analysis showed that NB-MSCs had an increased number of cells in the G0-G1 phase compared to BM-MSCs. Transcriptomic profiling results indicated that NB-MSCs were enriched with EMT genes compared to BM-MSCs. Conclusions We characterized the biological features, the immunomodulatory capacity and the gene expression profile of NB-MSCs. The NB-MSC gene expression profile and their functional properties suggest a potential role in promoting tumor escape, invasiveness and metastatic traits of NB cancer cells. A better understanding of the complex mechanisms underlying the interactions between NB cells and NB-derived MSCs should shed new light on potential novel therapeutic approaches.http://link.springer.com/article/10.1186/s12885-018-5082-2NeuroblastomaMesenchymal stromal cellsMicroenvironmentStemnessEMTChildren |
| spellingShingle | Gloria Pelizzo Veronica Veschi Melissa Mantelli Stefania Croce Vincenzo Di Benedetto Paolo D’Angelo Alice Maltese Laura Catenacci Tiziana Apuzzo Emanuela Scavo Antonia Moretta Matilde Todaro Giorgio Stassi Maria Antonietta Avanzini Valeria Calcaterra Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cells BMC Cancer Neuroblastoma Mesenchymal stromal cells Microenvironment Stemness EMT Children |
| title | Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cells |
| title_full | Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cells |
| title_fullStr | Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cells |
| title_full_unstemmed | Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cells |
| title_short | Microenvironment in neuroblastoma: isolation and characterization of tumor-derived mesenchymal stromal cells |
| title_sort | microenvironment in neuroblastoma isolation and characterization of tumor derived mesenchymal stromal cells |
| topic | Neuroblastoma Mesenchymal stromal cells Microenvironment Stemness EMT Children |
| url | http://link.springer.com/article/10.1186/s12885-018-5082-2 |
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