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HIF activation enhances FcγRIIb expression on mononuclear phagocytes impeding tumor targeting antibody immunotherapy

HIF activation enhances FcγRIIb expression on mononuclear phagocytes impeding tumor targeting antibody immunotherapy
HIF activation enhances FcγRIIb expression on mononuclear phagocytes impeding tumor targeting antibody immunotherapy
Background
Hypoxia is a hallmark of the tumor microenvironment (TME) and in addition to altering metabolism in cancer cells, it transforms tumor-associated stromal cells. Within the tumor stromal cell compartment, tumor-associated macrophages (TAMs) provide potent pro-tumoral support. However, TAMs can also be harnessed to destroy tumor cells by monoclonal antibody (mAb) immunotherapy, through antibody dependent cellular phagocytosis (ADCP). This is mediated via antibody-binding activating Fc gamma receptors (FcγR) and impaired by the single inhibitory FcγR, FcγRIIb.

Methods
We applied a multi-OMIC approach coupled with in vitro functional assays and murine tumor models to assess the effects of hypoxia inducible factor (HIF) activation on mAb mediated depletion of human and murine cancer cells. For mechanistic assessments, siRNA-mediated gene silencing, Western blotting and chromatin immune precipitation were utilized to assess the impact of identified regulators on FCGR2B gene transcription.

Results
We report that TAMs are FcγRIIbbright relative to healthy tissue counterparts and under hypoxic conditions, mononuclear phagocytes markedly upregulate FcγRIIb. This enhanced FcγRIIb expression is transcriptionally driven through HIFs and Activator protein 1 (AP-1). Importantly, this phenotype reduces the ability of macrophages to eliminate anti-CD20 monoclonal antibody (mAb) opsonized human chronic lymphocytic leukemia cells in vitro and EL4 lymphoma cells in vivo in human FcγRIIb+/+ transgenic mice. Furthermore, post-HIF activation, mAb mediated blockade of FcγRIIb can partially restore phagocytic function in human monocytes.

Conclusion
Our findings provide a detailed molecular and cellular basis for hypoxia driven resistance to antitumor mAb immunotherapy, unveiling a hitherto unexplored aspect of the TME. These findings provide a mechanistic rationale for the modulation of FcγRIIb expression or its blockade as a promising strategy to enhance approved and novel mAb immunotherapies.
0392-9078
Cragg, Mark
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Beers, Stephen
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Hussain, Khiyam
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Cleary, Kirstie
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Oldham, Robert
f43d1416-0b93-4dfd-a504-2850cb43e87e
Foxall, Russell
cfe3a818-a281-4bcb-8889-e1d0b591117c
James, Sonya
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Booth, Steven
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Longley, Jemma
f9f57245-1b80-44ee-8775-0893d1b383c3
Stopforth, Richard J
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Roghanian, Ali
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Carter, Matthew
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Cragg, Mark
ec97f80e-f3c8-49b7-a960-20dff648b78c
Beers, Stephen
a02548be-3ffd-41ab-9db8-d6e8c3b499a2
Hussain, Khiyam
9468f252-81d0-4251-b800-702433b610f8
Cleary, Kirstie
16e11432-9855-4b5b-8c3f-be86f0ef51f0
Oldham, Robert
f43d1416-0b93-4dfd-a504-2850cb43e87e
Foxall, Russell
cfe3a818-a281-4bcb-8889-e1d0b591117c
James, Sonya
764c80e3-5bea-4b34-a871-b43f87ef97b0
Booth, Steven
c2026d9d-ed93-4b1b-bce5-6b3efc8b8ca5
Longley, Jemma
f9f57245-1b80-44ee-8775-0893d1b383c3
Stopforth, Richard J
2d3e18ff-5563-4247-9150-f0f337fb585f
Roghanian, Ali
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Carter, Matthew
3baac102-d80c-42ba-ab4d-ad339a48169e

Cragg, Mark, Beers, Stephen, Hussain, Khiyam, Cleary, Kirstie, Oldham, Robert, Foxall, Russell, James, Sonya, Booth, Steven, Longley, Jemma, Stopforth, Richard J, Roghanian, Ali and Carter, Matthew (2022) HIF activation enhances FcγRIIb expression on mononuclear phagocytes impeding tumor targeting antibody immunotherapy. Journal of Experimental and Clinical Cancer Research, 41 (131). (doi:10.1186/s13046-022-02294-5).

Record type: Article

Abstract

Background
Hypoxia is a hallmark of the tumor microenvironment (TME) and in addition to altering metabolism in cancer cells, it transforms tumor-associated stromal cells. Within the tumor stromal cell compartment, tumor-associated macrophages (TAMs) provide potent pro-tumoral support. However, TAMs can also be harnessed to destroy tumor cells by monoclonal antibody (mAb) immunotherapy, through antibody dependent cellular phagocytosis (ADCP). This is mediated via antibody-binding activating Fc gamma receptors (FcγR) and impaired by the single inhibitory FcγR, FcγRIIb.

Methods
We applied a multi-OMIC approach coupled with in vitro functional assays and murine tumor models to assess the effects of hypoxia inducible factor (HIF) activation on mAb mediated depletion of human and murine cancer cells. For mechanistic assessments, siRNA-mediated gene silencing, Western blotting and chromatin immune precipitation were utilized to assess the impact of identified regulators on FCGR2B gene transcription.

Results
We report that TAMs are FcγRIIbbright relative to healthy tissue counterparts and under hypoxic conditions, mononuclear phagocytes markedly upregulate FcγRIIb. This enhanced FcγRIIb expression is transcriptionally driven through HIFs and Activator protein 1 (AP-1). Importantly, this phenotype reduces the ability of macrophages to eliminate anti-CD20 monoclonal antibody (mAb) opsonized human chronic lymphocytic leukemia cells in vitro and EL4 lymphoma cells in vivo in human FcγRIIb+/+ transgenic mice. Furthermore, post-HIF activation, mAb mediated blockade of FcγRIIb can partially restore phagocytic function in human monocytes.

Conclusion
Our findings provide a detailed molecular and cellular basis for hypoxia driven resistance to antitumor mAb immunotherapy, unveiling a hitherto unexplored aspect of the TME. These findings provide a mechanistic rationale for the modulation of FcγRIIb expression or its blockade as a promising strategy to enhance approved and novel mAb immunotherapies.

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More information

Accepted/In Press date: 20 February 2022
Published date: 7 April 2022

Identifiers

Local EPrints ID: 456881
URI: http://eprints.soton.ac.uk/id/eprint/456881
ISSN: 0392-9078
PURE UUID: 3df779fe-a7e4-4ac7-b4e2-fa259a7f40b1
ORCID for Mark Cragg: ORCID iD orcid.org/0000-0003-2077-089X
ORCID for Stephen Beers: ORCID iD orcid.org/0000-0002-3765-3342
ORCID for Kirstie Cleary: ORCID iD orcid.org/0000-0001-6200-4945
ORCID for Richard J Stopforth: ORCID iD orcid.org/0000-0002-0054-7503
ORCID for Ali Roghanian: ORCID iD orcid.org/0000-0003-1316-4218

Catalogue record

Date deposited: 16 May 2022 16:30
Last modified: 17 May 2022 01:46

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Contributors

Author: Mark Cragg ORCID iD
Author: Stephen Beers ORCID iD
Author: Khiyam Hussain
Author: Kirstie Cleary ORCID iD
Author: Robert Oldham
Author: Russell Foxall
Author: Sonya James
Author: Steven Booth
Author: Jemma Longley
Author: Richard J Stopforth ORCID iD
Author: Ali Roghanian ORCID iD
Author: Matthew Carter

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