Abstract
The MYCN oncogene is a strong genetic marker associated with poor prognosis in neuroblastoma (NB). Therefore, MYCN gene amplification and subsequent overexpression provide a possible target for new treatment approaches in NB. We first identified an inverse correlation of MYCN expression with CD45 mRNA in 101 NB tumor samples. KEGG map** further revealed that MYCN expression was associated with immune-suppressive pathways characterized by a down-regulation of T cell activation and up-regulation of T cell inhibitory gene transcripts. We then aimed to investigate whether DNA vaccination against MYCN is effective to induce an antigen-specific and T cell-mediated immune response. For this purpose, we generated a MYCN-expressing syngeneic mouse model by MYCN gene transfer to NXS2 cells. MYCN-DNA vaccines were engineered based on the pCMV-F3Ub plasmid backbone to drive ubiquitinated full-length MYCN-cDNA and minigene expression. Vaccines were delivered orally with attenuated S. typhimurium strain SL7207 as a carrier. Immunization with both MYCN-DNA vaccines significantly reduced primary tumor growth of MYCN-expressing NB cells in contrast to negative controls. The immune response was mediated by tumor-infiltrating T cells in vivo, which revealed MYCN-specific and MHC class I-restricted lysis of inducible MYCN-expressing NB target cells in vitro. Finally, these antigen-specific T cells also killed MYCN-negative mammary carcinoma cells pulsed with MYCN peptides in contrast to controls. In summary, we demonstrate proof of concept that MYCN can be targeted by DNA vaccination, which may provide an approach to overcoming MYCN immune-suppressive activities in patients with MYCN-amplified disease.
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Abbreviations
- BET:
-
Extraterminal bromodomain Barone
- CAR:
-
Chimeric antigen receptors
- CD:
-
Cluster of differentiation
- cDNA:
-
Copy DNA
- CHO:
-
Chinese hamster ovary
- CTLs:
-
Cytotoxic T cells
- DOX:
-
Doxycycline
- ELISA:
-
Enzyme-linked immunosorbent assay
- FACS:
-
Fluorescence-activated cell sorting
- FCS:
-
Fetal calf serum
- gDNA:
-
Genomic DNA
- H:
-
Hour
- IFN:
-
Interferon
- KEGG:
-
Kyoto encyclopedia of genes and genomes
- LPS:
-
Lipopolysaccharide
- MHC:
-
Major histocompatibility complex
- MYCN:
-
Neuroblastoma-derived myelocytomatosis viral-related oncogene
- MNA:
-
MYCN amplification
- NB:
-
Neuroblastoma
- PCR:
-
Polymerase chain reaction
- RT:
-
Room temperature
- SD:
-
Standard deviation
- TAA:
-
Tumor-associated antigen
- TBS:
-
Tris-buffered saline
- TH:
-
Tyrosine hydroxylase
- TMA:
-
Tissue microarray
- wt:
-
Wild type
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Acknowledgments
This work was supported by NGFNplus (Bundesministerium für Bildung und Forschung, ENGINE).
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We have no conflict of interest to disclose.
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All applicable international, national and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.
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Stermann, A., Huebener, N., Seidel, D. et al. Targeting of MYCN by means of DNA vaccination is effective against neuroblastoma in mice. Cancer Immunol Immunother 64, 1215–1227 (2015). https://doi.org/10.1007/s00262-015-1733-1
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DOI: https://doi.org/10.1007/s00262-015-1733-1