Immune characterization of pre-clinical murine models of neuroblastoma
Immune characterization of pre-clinical murine models of neuroblastoma
Immunotherapy offers a potentially less toxic, more tumor-specific treatment for neuroblastoma than conventional cytotoxic therapies. Accurate and reproducible immune competent preclinical models are key to understanding mechanisms of action, interactions with other therapies and mechanisms of resistance to immunotherapy. Here we characterized the tumor and splenic microenvironment of two syngeneic subcutaneous (NXS2 and 9464D), and a spontaneous transgenic (TH-MYCN) murine model of neuroblastoma, comparing histological features and immune infiltrates to previously published data on human neuroblastoma. Histological sections of frozen tissues were stained by immunohistochemistry and immunofluorescence for immune cell markers and tumor architecture. Tissues were dissociated by enzymatic digestion, stained with panels of antibodies to detect and quantify cancer cells, along with lymphocytic and myeloid infiltration by flow cytometry. Finally, we tested TH-MYCN mice as a feasible model for immunotherapy, using prior treatment with cyclophosphamide to create a therapeutic window of minimal residual disease to favor host immune development. Immune infiltration differed significantly between all the models. TH-MYCN tumors were found to resemble immune infiltration in human tumors more closely than the subcutaneous models, alongside similar GD2 and MHC class I expression. Finally, TH-MYCN transgenic mice were administered cyclophosphamide alone or in combination with an anti-GD2 or anti-4-1BB monoclonal antibody, which resulted in increase in survival in both combination therapies. The TH-MYCN transgenic mouse is a promising in vivo model for testing immunotherapy compounds and combination therapy in a preclinical setting.
Webb, Emily
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Lanati, Silvia
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Wareham, Carol
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Easton, Alistair
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Dunn, Stuart N.
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Inzhelevskaya, Tatyana
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Sadler, Freja M.
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James, Sonya
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Ashton-Key, Margaret
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Cragg, Mark
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Beers, Stephen
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Gray, Juliet
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7 October 2020
Webb, Emily
013d6243-21c9-4d92-aab5-a06c838b256b
Lanati, Silvia
a614cb92-0753-4633-b7c7-acf67c512343
Wareham, Carol
ae0b8cb2-7887-4541-ac48-60a5bdcd4241
Easton, Alistair
d3eccbc8-2f4d-41af-bc9d-9d8c29130d7d
Dunn, Stuart N.
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Inzhelevskaya, Tatyana
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Sadler, Freja M.
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James, Sonya
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Ashton-Key, Margaret
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Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Beers, Stephen
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Gray, Juliet
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Webb, Emily, Lanati, Silvia, Wareham, Carol, Easton, Alistair, Dunn, Stuart N., Inzhelevskaya, Tatyana, Sadler, Freja M., James, Sonya, Ashton-Key, Margaret, Cragg, Mark, Beers, Stephen and Gray, Juliet
(2020)
Immune characterization of pre-clinical murine models of neuroblastoma.
Scientific Reports, 10 (1), [16695].
(doi:10.1038/s41598-020-73695-9).
Abstract
Immunotherapy offers a potentially less toxic, more tumor-specific treatment for neuroblastoma than conventional cytotoxic therapies. Accurate and reproducible immune competent preclinical models are key to understanding mechanisms of action, interactions with other therapies and mechanisms of resistance to immunotherapy. Here we characterized the tumor and splenic microenvironment of two syngeneic subcutaneous (NXS2 and 9464D), and a spontaneous transgenic (TH-MYCN) murine model of neuroblastoma, comparing histological features and immune infiltrates to previously published data on human neuroblastoma. Histological sections of frozen tissues were stained by immunohistochemistry and immunofluorescence for immune cell markers and tumor architecture. Tissues were dissociated by enzymatic digestion, stained with panels of antibodies to detect and quantify cancer cells, along with lymphocytic and myeloid infiltration by flow cytometry. Finally, we tested TH-MYCN mice as a feasible model for immunotherapy, using prior treatment with cyclophosphamide to create a therapeutic window of minimal residual disease to favor host immune development. Immune infiltration differed significantly between all the models. TH-MYCN tumors were found to resemble immune infiltration in human tumors more closely than the subcutaneous models, alongside similar GD2 and MHC class I expression. Finally, TH-MYCN transgenic mice were administered cyclophosphamide alone or in combination with an anti-GD2 or anti-4-1BB monoclonal antibody, which resulted in increase in survival in both combination therapies. The TH-MYCN transgenic mouse is a promising in vivo model for testing immunotherapy compounds and combination therapy in a preclinical setting.
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Accepted/In Press date: 18 September 2020
e-pub ahead of print date: 7 October 2020
Published date: 7 October 2020
Additional Information:
Funding Information:
E.R.W was funded by Neuroblastoma UK and a University of Southampton Vice Chancellor’s Scholarship. S.L was funded by Sparks Children’s Medical Research Charity. C.W was funded by The Dubois Children’s Cancer Fund. This work was also funded by Cancer Research UK (C1477/A10834, C8574/A11781 and C34999A/ A18087). No funders had any role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript.
Publisher Copyright:
© 2020, The Author(s).
Identifiers
Local EPrints ID: 444286
URI: http://eprints.soton.ac.uk/id/eprint/444286
ISSN: 2045-2322
PURE UUID: 9fd2d348-5a63-4bb0-a0de-e0d77ba23996
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Date deposited: 08 Oct 2020 16:51
Last modified: 17 Mar 2024 02:57
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Contributors
Author:
Emily Webb
Author:
Carol Wareham
Author:
Alistair Easton
Author:
Stuart N. Dunn
Author:
Tatyana Inzhelevskaya
Author:
Freja M. Sadler
Author:
Sonya James
Author:
Margaret Ashton-Key
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