Macrophage Plasticity and Function in the Lung Tumour Microenvironment Revealed in 3D Heterotypic Spheroid and Explant Models
In non-small cell lung cancer (NSCLC), stroma-resident and tumour-infiltrating macrophages may facilitate an immunosuppressive tumour microenvironment (TME) and hamper immunotherapeutic responses. Analysis of tumour-associated macrophage (TAM) plasticity in NSCLC is largely lacking. We established a...
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| Format: | Article |
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MDPI AG
2021-03-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/9/3/302 |
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| author | Lauren Evans Kate Milward Richard Attanoos Aled Clayton Rachel Errington Zsuzsanna Tabi |
| author_facet | Lauren Evans Kate Milward Richard Attanoos Aled Clayton Rachel Errington Zsuzsanna Tabi |
| author_sort | Lauren Evans |
| collection | DOAJ |
| description | In non-small cell lung cancer (NSCLC), stroma-resident and tumour-infiltrating macrophages may facilitate an immunosuppressive tumour microenvironment (TME) and hamper immunotherapeutic responses. Analysis of tumour-associated macrophage (TAM) plasticity in NSCLC is largely lacking. We established a novel, multi-marker, dual analysis approach for assessing monocyte-derived macrophage (Mφ) polarisation and M1/M2 phenotypic plasticity. We developed a flow cytometry-based, two-marker analysis (CD64 and CD206) of CD14<sup>+</sup> cells. The phenotype and immune function of in vitro-induced TAMs was studied in a heterotypic spheroid and tumour-derived explant model of NSCLC. Heterotypic spheroids and NSCLC explants skewed Mφs from an M1- (CD206<sup>lo</sup>CD64<sup>hi</sup>) to M2-like (CD206<sup>hi</sup>CD64<sup>lo</sup>) phenotype. Lipopolysaccharide (LPS) and IFNγ treatment reversed M2-like Mφ polarisation, indicating the plasticity of Mφs. Importantly, antigen-specific CD8<sup>+</sup> T cell responses were reduced in the presence of tumour explant-conditioned Mφs, but not spheroid-conditioned Mφs, suggesting explants are likely a more relevant model of the immune TME than cell line-derived spheroids. Our data indicates the importance of multi-marker, functional analyses within Mφ subsets and the advantages of the ex vivo NSCLC explant model in immunomodulation studies. We highlight the plasticity of the M1/M2 phenotype using the explant model and provide a tool for studying therapeutic interventions designed to reprogram M2-like Mφ-induced immunosuppression. |
| first_indexed | 2024-03-10T13:13:39Z |
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| language | English |
| last_indexed | 2024-03-10T13:13:39Z |
| publishDate | 2021-03-01 |
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| series | Biomedicines |
| spelling | doaj.art-94b8b71357ef4920a206ae84680e22882023-11-21T10:36:04ZengMDPI AGBiomedicines2227-90592021-03-019330210.3390/biomedicines9030302Macrophage Plasticity and Function in the Lung Tumour Microenvironment Revealed in 3D Heterotypic Spheroid and Explant ModelsLauren Evans0Kate Milward1Richard Attanoos2Aled Clayton3Rachel Errington4Zsuzsanna Tabi5Tissue MicroEnvironment Group, Division of Cancer and Genetics, School of Medicine, Cardiff University, Tenovus Building, University Hospital of Wales, Heath Park, Cardiff CF14 4XN, UKTissue MicroEnvironment Group, Division of Cancer and Genetics, School of Medicine, Cardiff University, Tenovus Building, University Hospital of Wales, Heath Park, Cardiff CF14 4XN, UKSchool of Medicine, Cardiff University, University Hospital of Wales, Heath Park, Cardiff CF14 4XN, UKTissue MicroEnvironment Group, Division of Cancer and Genetics, School of Medicine, Cardiff University, Tenovus Building, University Hospital of Wales, Heath Park, Cardiff CF14 4XN, UKTissue MicroEnvironment Group, Division of Cancer and Genetics, School of Medicine, Cardiff University, Tenovus Building, University Hospital of Wales, Heath Park, Cardiff CF14 4XN, UKTissue MicroEnvironment Group, Division of Cancer and Genetics, School of Medicine, Cardiff University, Tenovus Building, University Hospital of Wales, Heath Park, Cardiff CF14 4XN, UKIn non-small cell lung cancer (NSCLC), stroma-resident and tumour-infiltrating macrophages may facilitate an immunosuppressive tumour microenvironment (TME) and hamper immunotherapeutic responses. Analysis of tumour-associated macrophage (TAM) plasticity in NSCLC is largely lacking. We established a novel, multi-marker, dual analysis approach for assessing monocyte-derived macrophage (Mφ) polarisation and M1/M2 phenotypic plasticity. We developed a flow cytometry-based, two-marker analysis (CD64 and CD206) of CD14<sup>+</sup> cells. The phenotype and immune function of in vitro-induced TAMs was studied in a heterotypic spheroid and tumour-derived explant model of NSCLC. Heterotypic spheroids and NSCLC explants skewed Mφs from an M1- (CD206<sup>lo</sup>CD64<sup>hi</sup>) to M2-like (CD206<sup>hi</sup>CD64<sup>lo</sup>) phenotype. Lipopolysaccharide (LPS) and IFNγ treatment reversed M2-like Mφ polarisation, indicating the plasticity of Mφs. Importantly, antigen-specific CD8<sup>+</sup> T cell responses were reduced in the presence of tumour explant-conditioned Mφs, but not spheroid-conditioned Mφs, suggesting explants are likely a more relevant model of the immune TME than cell line-derived spheroids. Our data indicates the importance of multi-marker, functional analyses within Mφ subsets and the advantages of the ex vivo NSCLC explant model in immunomodulation studies. We highlight the plasticity of the M1/M2 phenotype using the explant model and provide a tool for studying therapeutic interventions designed to reprogram M2-like Mφ-induced immunosuppression.https://www.mdpi.com/2227-9059/9/3/302heterotypic spheroidsmyeloid cellsNSCLCtumour explantstumour microenvironment |
| spellingShingle | Lauren Evans Kate Milward Richard Attanoos Aled Clayton Rachel Errington Zsuzsanna Tabi Macrophage Plasticity and Function in the Lung Tumour Microenvironment Revealed in 3D Heterotypic Spheroid and Explant Models Biomedicines heterotypic spheroids myeloid cells NSCLC tumour explants tumour microenvironment |
| title | Macrophage Plasticity and Function in the Lung Tumour Microenvironment Revealed in 3D Heterotypic Spheroid and Explant Models |
| title_full | Macrophage Plasticity and Function in the Lung Tumour Microenvironment Revealed in 3D Heterotypic Spheroid and Explant Models |
| title_fullStr | Macrophage Plasticity and Function in the Lung Tumour Microenvironment Revealed in 3D Heterotypic Spheroid and Explant Models |
| title_full_unstemmed | Macrophage Plasticity and Function in the Lung Tumour Microenvironment Revealed in 3D Heterotypic Spheroid and Explant Models |
| title_short | Macrophage Plasticity and Function in the Lung Tumour Microenvironment Revealed in 3D Heterotypic Spheroid and Explant Models |
| title_sort | macrophage plasticity and function in the lung tumour microenvironment revealed in 3d heterotypic spheroid and explant models |
| topic | heterotypic spheroids myeloid cells NSCLC tumour explants tumour microenvironment |
| url | https://www.mdpi.com/2227-9059/9/3/302 |
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