Developing a 3D B cell Lymphoma culture system to model antibody therapy
Developing a 3D B cell Lymphoma culture system to model antibody therapy
Diffuse large cell B cell lymphoma (DLBCL) accounts for approximately 30%–40% of all non-Hodgkin lymphoma (NHL) cases. Current first line DLBCL treatment results in long-term remission in more than 60% of cases. However, those patients with primary refractory disease or early relapse exhibit poor prognosis, highlighting a requirement for alternative therapies. Our aim was to develop a novel model of DLBCL that facilitates in vitro testing of current and novel therapies by replicating key components of the tumor microenvironment (TME) in a three-dimensional (3D) culture system that would enable primary DLBCL cell survival and study ex vivo. The TME is a complex ecosystem, comprising malignant and non-malignant cells, including cancer-associated fibroblasts (CAF) and tumor-associated macrophages (TAM) whose reciprocal crosstalk drives tumor initiation and growth while fostering an immunosuppressive milieu enabling its persistence. The requirement to recapitulate, at least to some degree, this complex, interactive network is exemplified by the rapid cell death of primary DLBCL cells removed from their TME and cultured alone in vitro. Building on previously described methodologies to generate lymphoid-like fibroblasts from adipocyte derived stem cells (ADSC), we confirmed lymphocytes, specifically B cells, interacted with this ADSC-derived stroma, in the presence or absence of monocyte-derived macrophages (MDM), in both two-dimensional (2D) cultures and a 3D collagen-based spheroid system. Furthermore, we demonstrated that DLBCL cells cultured in this system interact with its constituent components, resulting in their improved viability as compared to ex-vivo 2D monocultures. We then assessed the utility of this system as a platform to study therapeutics in the context of antibody-directed phagocytosis, using rituximab as a model immunotherapeutic antibody. Overall, we describe a novel 3D spheroid co-culture system comprising key components of the DLBCL TME with the potential to serve as a testbed for novel therapeutics, targeting key cellular constituents of the TME, such as CAF and/or TAM.
3D co-culture model, adipocyte derived stem cell, antibody therapy, cancer associated fibroblast, diffuse large B cell lymphoma, tumor associated macrophage
Foxall, Russell
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Narang, Priyanka
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Glaysher, Bridget
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Hub, Elin
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Teal, Emma Louise
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Coles, Mark C.
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Ashton-Key, Margaret
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Beers, Stephen
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Cragg, Mark
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8 February 2021
Foxall, Russell
cfe3a818-a281-4bcb-8889-e1d0b591117c
Narang, Priyanka
b2e8fcbd-c75a-444b-b95f-39dea0bc5d3d
Glaysher, Bridget
a5186296-148c-476c-8bb9-965047c9271c
Hub, Elin
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Teal, Emma Louise
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Coles, Mark C.
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Ashton-Key, Margaret
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Beers, Stephen
a02548be-3ffd-41ab-9db8-d6e8c3b499a2
Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Foxall, Russell, Narang, Priyanka, Glaysher, Bridget, Hub, Elin, Teal, Emma Louise, Coles, Mark C., Ashton-Key, Margaret, Beers, Stephen and Cragg, Mark
(2021)
Developing a 3D B cell Lymphoma culture system to model antibody therapy.
Frontiers in Immunology, 11, [605231].
(doi:10.3389/fimmu.2020.605231).
Abstract
Diffuse large cell B cell lymphoma (DLBCL) accounts for approximately 30%–40% of all non-Hodgkin lymphoma (NHL) cases. Current first line DLBCL treatment results in long-term remission in more than 60% of cases. However, those patients with primary refractory disease or early relapse exhibit poor prognosis, highlighting a requirement for alternative therapies. Our aim was to develop a novel model of DLBCL that facilitates in vitro testing of current and novel therapies by replicating key components of the tumor microenvironment (TME) in a three-dimensional (3D) culture system that would enable primary DLBCL cell survival and study ex vivo. The TME is a complex ecosystem, comprising malignant and non-malignant cells, including cancer-associated fibroblasts (CAF) and tumor-associated macrophages (TAM) whose reciprocal crosstalk drives tumor initiation and growth while fostering an immunosuppressive milieu enabling its persistence. The requirement to recapitulate, at least to some degree, this complex, interactive network is exemplified by the rapid cell death of primary DLBCL cells removed from their TME and cultured alone in vitro. Building on previously described methodologies to generate lymphoid-like fibroblasts from adipocyte derived stem cells (ADSC), we confirmed lymphocytes, specifically B cells, interacted with this ADSC-derived stroma, in the presence or absence of monocyte-derived macrophages (MDM), in both two-dimensional (2D) cultures and a 3D collagen-based spheroid system. Furthermore, we demonstrated that DLBCL cells cultured in this system interact with its constituent components, resulting in their improved viability as compared to ex-vivo 2D monocultures. We then assessed the utility of this system as a platform to study therapeutics in the context of antibody-directed phagocytosis, using rituximab as a model immunotherapeutic antibody. Overall, we describe a novel 3D spheroid co-culture system comprising key components of the DLBCL TME with the potential to serve as a testbed for novel therapeutics, targeting key cellular constituents of the TME, such as CAF and/or TAM.
Text
2020-0312720-3
- Accepted Manuscript
More information
Accepted/In Press date: 16 December 2020
e-pub ahead of print date: 8 February 2021
Published date: 8 February 2021
Additional Information:
Funding Information:
We are grateful to Jenny Norman in the Histochemistry Research Unit, University Hospital Southampton, Maria Fernandes, in the Research Pathology, University Hospital Southampton and Dave Johnston in the Biomedical Imaging Unit, University Hospital Southampton for the assistance they provided.
Publisher Copyright:
© Copyright © 2021 Foxall, Narang, Glaysher, Hub, Teal, Coles, Ashton-Key, Beers and Cragg.
Keywords:
3D co-culture model, adipocyte derived stem cell, antibody therapy, cancer associated fibroblast, diffuse large B cell lymphoma, tumor associated macrophage
Identifiers
Local EPrints ID: 446842
URI: http://eprints.soton.ac.uk/id/eprint/446842
ISSN: 1664-3224
PURE UUID: f1f93cb9-e650-46a1-89b4-4b77852a332d
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Date deposited: 24 Feb 2021 17:31
Last modified: 17 Mar 2024 02:52
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Contributors
Author:
Russell Foxall
Author:
Priyanka Narang
Author:
Bridget Glaysher
Author:
Elin Hub
Author:
Emma Louise Teal
Author:
Mark C. Coles
Author:
Margaret Ashton-Key
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