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Immune characterization of pre-clinical murine models of neuroblastoma

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.
2045-2322
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
2d84ec97-18c9-4406-a96b-206c21eae7d5
Sadler, Freja M.
James, Sonya
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Ashton-Key, Margaret
5111ac18-7d4f-4ef0-9c71-0a44c37aaed4
Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Beers, Stephen
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Gray, Juliet
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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
2d84ec97-18c9-4406-a96b-206c21eae7d5
Sadler, Freja M.
James, Sonya
764c80e3-5bea-4b34-a871-b43f87ef97b0
Ashton-Key, Margaret
5111ac18-7d4f-4ef0-9c71-0a44c37aaed4
Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Beers, Stephen
a02548be-3ffd-41ab-9db8-d6e8c3b499a2
Gray, Juliet
12d5e17c-97bb-4d6d-8fc4-3914b730ed42

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, [16695]. (doi:10.1038/s41598-020-73695-9).

Record type: Article

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

Identifiers

Local EPrints ID: 444286
URI: http://eprints.soton.ac.uk/id/eprint/444286
ISSN: 2045-2322
PURE UUID: 9fd2d348-5a63-4bb0-a0de-e0d77ba23996
ORCID for Mark Cragg: ORCID iD orcid.org/0000-0003-2077-089X

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Date deposited: 08 Oct 2020 16:51
Last modified: 08 Oct 2020 16:51

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Contributors

Author: Emily Webb
Author: Silvia Lanati
Author: Carol Wareham
Author: Alistair Easton
Author: Stuart N. Dunn
Author: Tatyana Inzhelevskaya
Author: Freja M. Sadler
Author: Sonya James
Author: Margaret Ashton-Key
Author: Mark Cragg ORCID iD
Author: Stephen Beers
Author: Juliet Gray

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