Cell-Laden Hydrogel as a Clinical-Relevant 3D Model for Analyzing Neuroblastoma Growth, Immunophenotype, and Susceptibility to Therapies
High risk Neuroblastoma (NB) includes aggressive, metastatic solid tumors of childhood. The survival rate improved only modestly, despite the use of combination therapies including novel immunotherapies based on the antibody-mediated targeting of tumor-associated surface ligands. Treatment failures...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-08-01
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| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2019.01876/full |
| _version_ | 1811324391942782976 |
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| author | Alessandra Marrella Alessandra Dondero Maurizio Aiello Beatrice Casu Daniel Olive Stefano Regis Cristina Bottino Cristina Bottino Daniela Pende Raffaella Meazza Guido Caluori Guido Caluori Roberta Castriconi Roberta Castriconi Silvia Scaglione Silvia Scaglione |
| author_facet | Alessandra Marrella Alessandra Dondero Maurizio Aiello Beatrice Casu Daniel Olive Stefano Regis Cristina Bottino Cristina Bottino Daniela Pende Raffaella Meazza Guido Caluori Guido Caluori Roberta Castriconi Roberta Castriconi Silvia Scaglione Silvia Scaglione |
| author_sort | Alessandra Marrella |
| collection | DOAJ |
| description | High risk Neuroblastoma (NB) includes aggressive, metastatic solid tumors of childhood. The survival rate improved only modestly, despite the use of combination therapies including novel immunotherapies based on the antibody-mediated targeting of tumor-associated surface ligands. Treatment failures may be due to the lack of adequate in vitro models for studying, in a given patient, the efficacy of potential therapeutics, including those aimed to enhance anti-tumor immune responses. We here propose a 3D alginate-based hydrogel as extracellular microenvironment to evaluate the effects of the three-dimensionality on biological and immunological properties of NB cells. NB cell lines grown within the 3D alginate spheres presented spheroid morphology, optimal survival, and proliferation capabilities, and a reduced sensitivity to the cytotoxic effect of imatinib mesylate. 3D cultured NB cells were also evaluated for the constitutive and IFN-γ-induced expression of surface molecules capable of tuning the anti-tumor activity of NK cells including immune checkpoint ligands. In particular, IFN-γ induced de novo expression of high amounts of HLA-I molecules, which protected NB cells from the attack mediated by KIR/KIR-L matched NK cells. Moreover, in the 3D alginate spheres, the cytokine increased the expression of the immune checkpoint ligands PD-Ls and B7-H3 while virtually abrogating that of PVR, a ligand of DNAM-1 activating receptor, whose expression correlates with high susceptibility to NK-mediated killing. Our 3D model highlighted molecular features that more closely resemble the immunophenotypic variants occurring in vivo and not fully appreciated in classical 2D culture conditions. Thus, based on our results, 3D alginate-based hydrogels might represent a clinical-relevant cell culture platform where to test the efficacy of personalized therapeutic approaches aimed to optimize the current and innovative immune based therapies in a very systematic and reliable way. |
| first_indexed | 2024-04-13T14:13:26Z |
| format | Article |
| id | doaj.art-a38c4171e38547d9b6023846f4c5d30a |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-13T14:13:26Z |
| publishDate | 2019-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-a38c4171e38547d9b6023846f4c5d30a2022-12-22T02:43:43ZengFrontiers Media S.A.Frontiers in Immunology1664-32242019-08-011010.3389/fimmu.2019.01876457284Cell-Laden Hydrogel as a Clinical-Relevant 3D Model for Analyzing Neuroblastoma Growth, Immunophenotype, and Susceptibility to TherapiesAlessandra Marrella0Alessandra Dondero1Maurizio Aiello2Beatrice Casu3Daniel Olive4Stefano Regis5Cristina Bottino6Cristina Bottino7Daniela Pende8Raffaella Meazza9Guido Caluori10Guido Caluori11Roberta Castriconi12Roberta Castriconi13Silvia Scaglione14Silvia Scaglione15CNR—IEIIT Institute, National Research Council of Italy, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyReact4life S.r.l., Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyTumor Immunology Team, IBISA Immunomonitoring Platform, Cancer Research Center of Marseille, INSERM U1068, CNRS U7258, Institut Paoli-Calmettes, Aix-Marseille University, Marseille, FranceLaboratory of Clinical and Experimental Immunology, IRCCS Giannina Gaslini, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyLaboratory of Clinical and Experimental Immunology, IRCCS Giannina Gaslini, Genoa, ItalyLaboratorio di Immunologia, IRCCS Ospedale Policlinico San Martino, Genoa, ItalyLaboratorio di Immunologia, IRCCS Ospedale Policlinico San Martino, Genoa, ItalyFNUSA-ICRC, Interventional Cardiac Electrophysiology, Brno, CzechiaNanobiotechnology, CEITEC Masaryk University, Brno, CzechiaDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyCentre of Excellence for Biomedical Research, CEBR, University of Genoa, Genoa, ItalyCNR—IEIIT Institute, National Research Council of Italy, Genoa, ItalyReact4life S.r.l., Genoa, ItalyHigh risk Neuroblastoma (NB) includes aggressive, metastatic solid tumors of childhood. The survival rate improved only modestly, despite the use of combination therapies including novel immunotherapies based on the antibody-mediated targeting of tumor-associated surface ligands. Treatment failures may be due to the lack of adequate in vitro models for studying, in a given patient, the efficacy of potential therapeutics, including those aimed to enhance anti-tumor immune responses. We here propose a 3D alginate-based hydrogel as extracellular microenvironment to evaluate the effects of the three-dimensionality on biological and immunological properties of NB cells. NB cell lines grown within the 3D alginate spheres presented spheroid morphology, optimal survival, and proliferation capabilities, and a reduced sensitivity to the cytotoxic effect of imatinib mesylate. 3D cultured NB cells were also evaluated for the constitutive and IFN-γ-induced expression of surface molecules capable of tuning the anti-tumor activity of NK cells including immune checkpoint ligands. In particular, IFN-γ induced de novo expression of high amounts of HLA-I molecules, which protected NB cells from the attack mediated by KIR/KIR-L matched NK cells. Moreover, in the 3D alginate spheres, the cytokine increased the expression of the immune checkpoint ligands PD-Ls and B7-H3 while virtually abrogating that of PVR, a ligand of DNAM-1 activating receptor, whose expression correlates with high susceptibility to NK-mediated killing. Our 3D model highlighted molecular features that more closely resemble the immunophenotypic variants occurring in vivo and not fully appreciated in classical 2D culture conditions. Thus, based on our results, 3D alginate-based hydrogels might represent a clinical-relevant cell culture platform where to test the efficacy of personalized therapeutic approaches aimed to optimize the current and innovative immune based therapies in a very systematic and reliable way.https://www.frontiersin.org/article/10.3389/fimmu.2019.01876/full3D cancer modelneuroblastomaNK cellsPVRKIRS/HLA-IPD-Ls |
| spellingShingle | Alessandra Marrella Alessandra Dondero Maurizio Aiello Beatrice Casu Daniel Olive Stefano Regis Cristina Bottino Cristina Bottino Daniela Pende Raffaella Meazza Guido Caluori Guido Caluori Roberta Castriconi Roberta Castriconi Silvia Scaglione Silvia Scaglione Cell-Laden Hydrogel as a Clinical-Relevant 3D Model for Analyzing Neuroblastoma Growth, Immunophenotype, and Susceptibility to Therapies Frontiers in Immunology 3D cancer model neuroblastoma NK cells PVR KIRS/HLA-I PD-Ls |
| title | Cell-Laden Hydrogel as a Clinical-Relevant 3D Model for Analyzing Neuroblastoma Growth, Immunophenotype, and Susceptibility to Therapies |
| title_full | Cell-Laden Hydrogel as a Clinical-Relevant 3D Model for Analyzing Neuroblastoma Growth, Immunophenotype, and Susceptibility to Therapies |
| title_fullStr | Cell-Laden Hydrogel as a Clinical-Relevant 3D Model for Analyzing Neuroblastoma Growth, Immunophenotype, and Susceptibility to Therapies |
| title_full_unstemmed | Cell-Laden Hydrogel as a Clinical-Relevant 3D Model for Analyzing Neuroblastoma Growth, Immunophenotype, and Susceptibility to Therapies |
| title_short | Cell-Laden Hydrogel as a Clinical-Relevant 3D Model for Analyzing Neuroblastoma Growth, Immunophenotype, and Susceptibility to Therapies |
| title_sort | cell laden hydrogel as a clinical relevant 3d model for analyzing neuroblastoma growth immunophenotype and susceptibility to therapies |
| topic | 3D cancer model neuroblastoma NK cells PVR KIRS/HLA-I PD-Ls |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2019.01876/full |
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