Abstract
Purpose
In a previous study, protein tyrosine phosphatase non-receptor type (PTPN) 3 was identified as an immune checkpoint molecule in lymphocytes, and its potential as a novel target for cancer immunotherapy was anticipated. However, evaluation of dendritic cell (DC) function as antigen-presenting cells is critical for the development of immunotherapy. In this study, we aimed to analyze the biological effect of PTPN3 on DCs induced from human peripheral blood monocytes obtained from healthy individuals.
Methods
We used short-interfering RNA to knock down PTP3 in DCs. For DC maturation, we added cancer cell lysate and tumor necrosis factor-α/interferon-α to immature DCs. In the cytotoxic assay, the target cancer cells were SBC5, unmatched with DCs from healthy human leukocyte antigen (HLA)-A24, or Sq-1, matched with DCs. Enzyme-linked immunosorbent assay was used to determine the amount of cytokines. To examine the intracellular signaling system, intracellular staining was used.
Results
PTPN3 knockdown significantly increased the number of DCs, expression of CD80 and chemokine receptor (CCR)7, and production of interleukin-12p40/p70 in mature DCs. In the HLA-A24-restricted DC and human lung squamous cell carcinoma cell cytotoxic assay, inhibition of PTPN3 expression in mature DCs induced cytotoxic T lymphocytes with increased production of INF-γ and granzyme B, and enhanced toxicity against cancer cells and migration to cancer. Furthermore, inhibition of PTPN3 expression activated the mitogen-activated protein kinase pathway in DCs.
Conclusion
Based on our findings, inhibition of PTPN3 expression could contribute to the development of novel cancer immunotherapies that activate not only lymphocytes but also DCs.
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Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- PTPN:
-
Protein tyrosine phosphatase non-receptor type
- DC:
-
Dendritic cell
- siRNA:
-
Short interfering RNA
- IL:
-
Interleukin
- HLA:
-
Human leukocyte antigen
- IFN:
-
Interferon
- MAPK:
-
Mitogen-activated protein kinase
- PD-1:
-
Programmed cell death receptor-1
- PD-L1:
-
Programmed cell death ligand-1
- CTLA-4:
-
Cytotoxic T-lymphocyte associated antigen-4
- LAG3:
-
Lymphocyte activation gene 3
- irAE:
-
Immune-related adverse events
- APC:
-
Antigen-presenting cell
- GM-CSF:
-
Granulocyte macrophage colony-stimulating factor
- PBMC:
-
Peripheral blood mononuclear cell
- TNF:
-
Tumor necrosis factor
- CCL:
-
Chemokine ligand
- FACS:
-
Fluorescence‑activated cell sorting
- FITC:
-
Fluorescein isothiocyanate
- PE:
-
Phycoerythrin
- PCR:
-
Polymerase chain reaction
- CCR:
-
Chemokine receptor
- CTLs:
-
Cytotoxic T lymphocytes
- NK cell:
-
Natural killer cell
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Acknowledgements
We would like to thank Ms. Emi Onishi for technical assistance. We also thank Editage for the English language review. This study was supported by the Japan Society for the Promotion of Science (KAKENHI Grant Number FAG21K08672).
Funding
This study was supported by the Japan Society for the Promotion of Science (KAKENHI Grant Number JP21K08672, JP21K09583, JP22H03163, JP22K08715).
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NI carried out the analysis of all experiments. SM, AI, KS, SM, SN, and SK were involved in the acquisition and analysis of data. HO, KO, MU, TM, and MN participated in the design of study. All the authors read and approved the final manuscript.
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The authors have no relevant financial or non-financial interests to disclose.
Ethical approval
All the procedures involving human participants performed in this study were in accordance with the ethical standards of the Kyushu University Ethics Committee (study approval numbers 29-251) and the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. All animal procedures were performed in accordance with the ethical standards of the Animal Care and Use Committee of Kyushu University (study approval number: A22-278-0).
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Written informed consent was obtained from all the participants included in the study for the use of their PBMC and tumor specimen for research and for publication before blood collection.
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Written informed consent was obtained from all participants included in the study for the use of their PBMC and tumor specimen for research and for publication before blood collection.
Animal source
All mice were obtained from Charles River Laboratories Japan (Yokohama, Japan).
Cell line authentication
SBC5 cell line was obtained from the Japanese Collection of Research Bioresources bank. The Sq-1 cell line was purchased from Riken Bio Resource Research Center. All cell lines were cultured for no more than 2–3 weeks after thawing, routinely checked for mycoplasma infection, and showed consistent phenotypes by microscopy prior to the in vitro and in vivo experiments.
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Iwamoto, N., Onishi, H., Masuda, S. et al. PTPN3 inhibition contributes to the activation of the dendritic cell function to be a promising new immunotherapy target. J Cancer Res Clin Oncol 149, 14619–14630 (2023). https://doi.org/10.1007/s00432-023-05250-8
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DOI: https://doi.org/10.1007/s00432-023-05250-8