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Targeting of MYCN by means of DNA vaccination is effective against neuroblastoma in mice

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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 mapping 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).

Conflict of interest

We have no conflict of interest to disclose.

Ethical approval

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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Authors and Affiliations

  1. Pediatric Hematology and Oncology, University Medicine Greifswald, Ferdinand-Sauerbruch-Str 1, 17475, Greifswald, Germany

    Alexander Stermann, Diana Seidel & Holger N. Lode

  2. Genetics of Metabolic and Reproductive Disorders, Max Delbrück Center for Molecular Medicine Berlin, Berlin, Germany

    Nicole Huebener

  3. Department of Pediatrics, Otto-von-Guericke University, Magdeburg, Germany

    Stefan Fest

  4. Department and Clinic of Pediatric Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany

    Georg Eschenburg

  5. Department of Pediatric Oncology and Hematology, University Children’s Hospital Essen, Essen, Germany

    Michael Stauder & Alexander Schramm

  6. Pediatric Oncology and Hematology, Charité University Medicine Berlin, Berlin, Germany

    Angelika Eggert

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  1. Alexander Stermann
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Correspondence to Holger N. Lode.

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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

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