Reprogramming Cancer Cells to Antigen-presenting Cells
Ferreira, Alexandra G LU
; Zimmermannova, Olga
LU
; Kurochkin, Ilia
LU
; Ascic, Ervin
LU
; Åkerström, Fritiof
LU
and Pereira, Carlos-Filipe
LU
(2023)
In Bio-protocol
13(22).
p.1-25
- Abstract
Cancer cells evade the immune system by downregulating antigen presentation. Although immune checkpoint inhibitors (ICI) and adoptive T-cell therapies revolutionized cancer treatment, their efficacy relies on the intrinsic immunogenicity of tumor cells and antigen presentation by dendritic cells. Here, we describe a protocol to directly reprogram murine and human cancer cells into tumor-antigen-presenting cells (tumor-APCs), using the type 1 conventional dendritic cell (cDC1) transcription factors PU.1, IRF8, and BATF3 delivered by a lentiviral vector. Tumor-APCs acquire a cDC1 cell-like phenotype, transcriptional and epigenetic programs, and function within nine days (Zimmermannova et al., 2023). Tumor-APCs express the hematopoietic... (More)
Cancer cells evade the immune system by downregulating antigen presentation. Although immune checkpoint inhibitors (ICI) and adoptive T-cell therapies revolutionized cancer treatment, their efficacy relies on the intrinsic immunogenicity of tumor cells and antigen presentation by dendritic cells. Here, we describe a protocol to directly reprogram murine and human cancer cells into tumor-antigen-presenting cells (tumor-APCs), using the type 1 conventional dendritic cell (cDC1) transcription factors PU.1, IRF8, and BATF3 delivered by a lentiviral vector. Tumor-APCs acquire a cDC1 cell-like phenotype, transcriptional and epigenetic programs, and function within nine days (Zimmermannova et al., 2023). Tumor-APCs express the hematopoietic marker CD45 and acquire the antigen presentation complexes MHC class I and II as well as co-stimulatory molecules required for antigen presentation to T cells, but do not express high levels of negative immune checkpoint regulators. Enriched tumor-APCs present antigens to Naïve CD8
(Less)
+ and CD4
+ T cells, are targeted by activated cytotoxic T lymphocytes, and elicit anti-tumor responses in vivo. The tumor-APC reprogramming protocol described here provides a simple and robust method to revert tumor evasion mechanisms by increasing antigen presentation in cancer cells. This platform has the potential to prime antigen-specific T-cell expansion, which can be leveraged for developing new cancer vaccines, neoantigen discovery, and expansion of tumor-infiltrating lymphocytes. Key features • This protocol describes the generation of antigen-presenting cells from cancer cells by direct reprogramming using lineage-instructive transcription factors of conventional dendritic cells type I. • Verification of reprogramming efficiency by flow cytometry and functional assessment of tumor-APCs by antigen presentation assays.
- author
- Ferreira, Alexandra G
LU
; Zimmermannova, Olga
LU
; Kurochkin, Ilia
LU
; Ascic, Ervin
LU
; Åkerström, Fritiof
LU
and Pereira, Carlos-Filipe
LU
- organization
- publishing date
- 2023-11-20
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Bio-protocol
- volume
- 13
- issue
- 22
- article number
- e4881
- pages
- 1 - 25
- publisher
- Bio-protocol LLC
- external identifiers
-
- scopus:85179045893
- pmid:38023788
- ISSN
- 2331-8325
- DOI
- 10.21769/BioProtoc.4881
- language
- English
- LU publication?
- yes
- additional info
- ©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license.
- id
- 074eb51a-7ced-4ed1-99e3-be7ef594083d
- date added to LUP
- 2023-12-06 20:14:52
- date last changed
- 2024-04-12 09:04:39
@article{074eb51a-7ced-4ed1-99e3-be7ef594083d,
abstract = {{<p>Cancer cells evade the immune system by downregulating antigen presentation. Although immune checkpoint inhibitors (ICI) and adoptive T-cell therapies revolutionized cancer treatment, their efficacy relies on the intrinsic immunogenicity of tumor cells and antigen presentation by dendritic cells. Here, we describe a protocol to directly reprogram murine and human cancer cells into tumor-antigen-presenting cells (tumor-APCs), using the type 1 conventional dendritic cell (cDC1) transcription factors PU.1, IRF8, and BATF3 delivered by a lentiviral vector. Tumor-APCs acquire a cDC1 cell-like phenotype, transcriptional and epigenetic programs, and function within nine days (Zimmermannova et al., 2023). Tumor-APCs express the hematopoietic marker CD45 and acquire the antigen presentation complexes MHC class I and II as well as co-stimulatory molecules required for antigen presentation to T cells, but do not express high levels of negative immune checkpoint regulators. Enriched tumor-APCs present antigens to Naïve CD8<br>
+ and CD4<br>
+ T cells, are targeted by activated cytotoxic T lymphocytes, and elicit anti-tumor responses in vivo. The tumor-APC reprogramming protocol described here provides a simple and robust method to revert tumor evasion mechanisms by increasing antigen presentation in cancer cells. This platform has the potential to prime antigen-specific T-cell expansion, which can be leveraged for developing new cancer vaccines, neoantigen discovery, and expansion of tumor-infiltrating lymphocytes. Key features • This protocol describes the generation of antigen-presenting cells from cancer cells by direct reprogramming using lineage-instructive transcription factors of conventional dendritic cells type I. • Verification of reprogramming efficiency by flow cytometry and functional assessment of tumor-APCs by antigen presentation assays.<br>
</p>}},
author = {{Ferreira, Alexandra G and Zimmermannova, Olga and Kurochkin, Ilia and Ascic, Ervin and Åkerström, Fritiof and Pereira, Carlos-Filipe}},
issn = {{2331-8325}},
language = {{eng}},
month = {{11}},
number = {{22}},
pages = {{1--25}},
publisher = {{Bio-protocol LLC}},
series = {{Bio-protocol}},
title = {{Reprogramming Cancer Cells to Antigen-presenting Cells}},
url = {{http://dx.doi.org/10.21769/BioProtoc.4881}},
doi = {{10.21769/BioProtoc.4881}},
volume = {{13}},
year = {{2023}},
}