A transplantable TH‐MYCN transgenic tumor model in C57Bl/6 mice for preclinical immunological studies in neuroblastoma

Kroesen, Michiel; Nierkens, Stefan; Ansems, Marleen; Wassink, Melissa; Orentas, Rimas J.; Boon, Louis; Brok, Martijn H. den; Hoogerbrugge, Peter M.; Adema, Gosse J.

doi:10.1002/ijc.28463

Cited by 51 publications

(74 citation statements)

References 47 publications

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“…Several of the models predominantly use immunocompromised mice and thus are unsuitable for assessment of immunotherapies, which are emerging as effective targeted therapies in patients with neuroblastoma. To overcome this, cell lines established from the TH-MYCN transgenic model/or other murine neuroblastoma cell lines can be used to generate orthotopic or pseudometastatic syngeneic models of neuroblastoma in an immunocompetent background (7,8). For this, the genetic and functional characterisation of murine cell lines, including Trp53 status and pathway function, are very important and highly warranted, as existing data are limited.…”

Section: Discussionmentioning

confidence: 99%

“…Although micrometastases can occur, one major limitation of the TH-MYCN transgenic model in preclinical drug development studies is the very low incidence of clinically relevant metastases to sites such as bone marrow, thus limiting its usefulness as a model for high-risk metastatic neuroblastoma (6). To overcome this, TH-MYCN cell lines have been used to generate highly valuable orthotopic and pseudometastatic syngeneic models of neuroblastoma in an immunocompetent background (7,8). This is of particular importance as immunotherapies, such as anti-GD 2 antibody are currently standard of care for the treatment of children with high-risk neuroblastoma.…”

Section: Introductionmentioning

confidence: 99%

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Characterisation of the p53 pathway in cell lines established from TH-MYCN transgenic mouse tumours

et al. 2018

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Cell lines established from the TH-MYCN transgenic murine model of neuroblastoma are a valuable preclinical, immunocompetent, syngeneic model of neuroblastoma, for which knowledge of their p53 pathway status is important. In this study, the Trp53 status and functional response to Nutlin-3 and ionising radiation (IR) were determined in 6 adherent TH-MYCN transgenic cell lines using Sanger sequencing, western blot analysis and flow cytometry. Sensitivity to structurally diverse MDM2 inhibitors (Nutlin-3, MI-63, RG7388 and NDD0005) was determined using XTT proliferation assays. In total, 2/6 cell lines were Trp53 homozygous mutant (NHO2A and 844MYCN+/+) and 1/6 (282MYCN+/-) was Trp53 heterozygous mutant. For 1/6 cell lines (NHO2A), DNA from the corresponding primary tumour was found to be Trp53 wt. In all cases, the presence of a mutation was consistent with aberrant p53 signalling in response to Nutlin-3 and IR. In comparison to TP53 wt human neuroblastoma cells, Trp53 wt murine control and TH-MYCN cell lines were significantly less sensitive to growth inhibition mediated by MI-63 and RG7388. These murine Trp53 wt and mutant TH-MYCN cell lines are useful syngeneic, immunocompetent neuroblastoma models, the former to test p53-dependent therapies in combination with immunotherapies, such as anti-GD2, and the latter as models of chemoresistant relapsed neuroblastoma when aberrations in the p53 pathway are more common. The spontaneous development of Trp53 mutations in 3 cell lines from TH-MYCN mice may have arisen from MYCN oncogenic driven and/or ex vivo selection. The identified species-dependent selectivity of MI-63 and RG7388 should be considered when interpreting in vivo toxicity studies of MDM2 inhibitors.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterisation of the p53 pathway in cell lines established from TH-MYCN transgenic mouse tumours

et al. 2018

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“…However, due to abdominally located tumors in the TH-MYCN mice, it is challenging to determine an objective endpoint to monitor the longitudinal effects of the treatment. Facilitated by imaging tools, several well-established murine models of neuroblastoma should be used, in order to further explore the mechanistic insights and longitudinal impact of combination immunotherapy (51)(52)(53)(54). Taken together, we conclude that tumor-driven CSF-1R signaling regulates the induction of suppressive myeloid cells, which hampers antitumor effects of checkpoint inhibitors.…”

Section: Discussionmentioning

confidence: 99%

Targeting Suppressive Myeloid Cells Potentiates Checkpoint Inhibitors to Control Spontaneous Neuroblastoma

Mao

Eißler

Blanc

et al. 2016

Clinical Cancer Research

128

105

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Purpose: Neuroblastoma is the most common extracranial solid cancer type in childhood, and high-risk patients have poor prognosis despite aggressive multimodal treatment. Neuroblastoma-driven inflammation contributes to the induction of suppressive myeloid cells that hamper efficient antitumor immune responses. Therefore, we sought to enhance antitumor immunity by removing immunosuppression mediated by myeloid cells.Experimental

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“…Neuro-2a cells were obtained from American Type Culture Collection (ATCC) (Manassas, Virginia). 9464D cells were derived from spontaneous TH-MYCN tumors 70 and obtained from Dr. Yonghzi Cui (National Cancer Institute). Cells were maintained in Dulbecco's modified Eagle's medium (GIBCO) supplemented with 10% fetal bovine serum (FBS; Invitrogen, Carlsbad, California) and 1% penicillin/streptomycin (GIBCO).…”

Section: Analysis Of Published Data Setsmentioning

confidence: 99%

Potent STING activation stimulates immunogenic cell death to enhance antitumor immunity in neuroblastoma

Wang-Bishop

Wehbe

Shae

et al. 2020

J Immunother Cancer

119

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BackgroundNeuroblastoma (NB) is a childhood cancer for which new treatment options are needed. The success of immune checkpoint blockade in the treatment of adult solid tumors has prompted the exploration of immunotherapy in NB; however, clinical evidence indicates that the vast majority of NB patients do not respond to single-agent checkpoint inhibitors. This motivates a need for therapeutic strategies to increase NB tumor immunogenicity. The goal of this study was to evaluate a new immunotherapeutic strategy for NB based on potent activation of the stimulator of interferon genes (STING) pathway.MethodsTo promote STING activation in NB cells and tumors, we utilized STING-activating nanoparticles (STING-NPs) that are designed to mediate efficient cytosolic delivery of the endogenous STING ligand, 2’3’-cGAMP. We investigated tumor-intrinsic responses to STING activation in both MYCN-amplified and non-amplified NB cell lines, evaluating effects on STING signaling, apoptosis, and the induction of immunogenic cell death. The effects of intratumoral administration of STING-NPs on CD8+T cell infiltration, tumor growth, and response to response to PD-L1 checkpoint blockade were evaluated in syngeneic models of MYCN-amplified and non-amplified NB.ResultsThe efficient cytosolic delivery of 2’3’-cGAMP enabled by STING-NPs triggered tumor-intrinsic STING signaling effects in both MYCN-amplified and non-amplified NB cell lines, resulting in increased expression of interferon-stimulated genes and pro-inflammatory cytokines as well as NB cell death at concentrations 2000-fold to 10000-fold lower than free 2’3’-cGAMP. STING-mediated cell death in NB was associated with release or expression of several danger associated molecular patterns that are hallmarks of immunogenic cell death, which was further validated via cell-based vaccination and tumor challenge studies. Intratumoral administration of STING-NPs enhanced STING activation relative to free 2’3’-cGAMP in NB tumor models, converting poorly immunogenic tumors into tumoricidal and T cell-inflamed microenvironments and resulting in inhibition of tumor growth, increased survival, and induction of immunological memory that protected against tumor re-challenge. In a model of MYCN-amplified NB, STING-NPs generated an abscopal response that inhibited distal tumor growth and improved response to PD-L1 immune checkpoint blockade.ConclusionsWe have demonstrated that activation of the STING pathway, here enabled by a nanomedicine approach, stimulates immunogenic cell death and remodels the tumor immune microenvironment to inhibit NB tumor growth and improve responses to immune checkpoint blockade, providing a multifaceted immunotherapeutic approach with potential to enhance immunotherapy outcomes in NB.

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A transplantable TH‐MYCN transgenic tumor model in C57Bl/6 mice for preclinical immunological studies in neuroblastoma

Cited by 51 publications

References 47 publications

Characterisation of the p53 pathway in cell lines established from TH-MYCN transgenic mouse tumours

Characterisation of the p53 pathway in cell lines established from TH-MYCN transgenic mouse tumours

Targeting Suppressive Myeloid Cells Potentiates Checkpoint Inhibitors to Control Spontaneous Neuroblastoma

Potent STING activation stimulates immunogenic cell death to enhance antitumor immunity in neuroblastoma

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