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Next-generation humanized patient-derived xenograft mouse model for pre-clinical antibody studies in neuroblastoma

Cancer Immunology, Immunotherapy volume 70, pages 721–732 (2021)Cite this article

Abstract

Faithful tumor mouse models are fundamental research tools to advance the field of immuno-oncology (IO). This is particularly relevant in diseases with low incidence, as in the case of pediatric malignancies, that rely on pre-clinical therapeutic development. However, conventional syngeneic and genetically engineered mouse models fail to recapitulate the tumor heterogeneity and microenvironmental complexity of human pathology that are essential determinants of cancer-directed immunity. Here, we characterize a novel mouse model that supports human natural killer (NK) cell development and engraftment of neuroblastoma orthotopic patient-derived xenograft (O-PDX) for pre-clinical antibody and cytokine testing. Using cytotoxicity assays, single-cell RNA-sequencing, and multi-color flow cytometry, we demonstrate that NK cells that develop in the humanized mice are fully licensed to execute NK cell cytotoxicity, permit human tumor engraftment, but can be therapeutically redirected to induce antibody-dependent cell-mediated cytotoxicity (ADCC). Although these cells share phenotypic and molecular features with healthy controls, we noted that they lacked an NK cell subset, termed activated NK cells, that is characterized by differentially expressed genes that are induced by cytokine activation. Because this subset of genes is also downregulated in patients with neuroblastoma compared to healthy controls, we hypothesize that this finding could be due to tumor-mediated suppressive effects. Thus, despite its technical complexity, this humanized patient-derived xenograft mouse model could serve as a faithful system for future testing of IO applications and studies of underlying immunologic processes.

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Fig. 1
Fig. 2

source of effector cells. Transplanted MISTRG mice (red) are compared with MISTRG (blue) and NSG (grey) mice that received ACT of NK cells from one donor. Among mice receiving chemo-immunotherapy, transplanted MISTRG mice (N = 7) had more significant tumor growth suppression than did MISTRG and NSG mice that received adoptive NK cells with therapy (N = 5 and 6; P = 0.003; one-way ANOVA). No significant differences occurred between these groups treated with chemotherapy (each MISTRG groups N = 6, NSG group N = 5)

Fig. 3
Fig. 4

Abbreviations

ACT:

Adoptive cell transfer

ADCC:

Antibody-dependent cell-mediated cytotoxicity

CAR:

Chimeric antigen receptor

E.T :

Effector-to-target

FACS:

Fluorescence-activated cell sorting

GD2:

Disialoganglioside

GEMM:

Genetically engineered mouse model

GM-CSF:

Granulocyte–macrophage colony-stimulating factor

HLA:

Human leukocyte antigen

HPCs:

Hematopoietic progenitor cells

IL:

Interleukin

IO:

Immuno-oncology

KIR:

Killer-cell immunoglobulin-like receptors

MISTRG:

M-CSFH/hIL-3/GM-CSFh/hSIRPAh/mTPOh/hRag2null/Il2Rγcnull

MITRG:

M-CSFH/hIL-3/GM-CSFh/hTPOh/hRag2null/Il2Rγcnull

NK:

Natural killer

NSG:

NOD/scid/Il2RγcNull

O-PDX:

Orthotopic patient-derived xenograft

scRNA-seq:

Single-cell RNA-sequencing

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Acknowledgements

We thank Asa Karlstrom, Ph.D., for regulatory support; Nisha Badders, Ph.D., ELS, for editing the manuscript; Mitchell J. Weiss, MD Ph.D., for providing HPCs; William E. Janssen, Ph.D., for providing purity data on HPCs; Jennifer Peters, Ph.D., for acquiring images of the stained tumor slides; Thomas Confer for assisting with the ultrasound studies; Merck Sorono and Children's GMP, LLC for providing hu14.18K322A for our studies; and the NCI Biological Resource Branch for providing IL-2 and IL-15. We thank Adeline Crinier, Bertrand Escalier, and Eric Vivier, Ph.D., for providing the biocomputational datasets of healthy controls. This work was supported, in part, by Cancer Center Support (CA21765) from the National Cancer Institute and grants to RN from the Conquer Cancer ASCO Foundation (12822), to WJA from the National Institutes of Health (R50CA211481), and to MAD from the National Institutes of Health (EY014867 and EY018599 and CA168875). This research was supported by Howard Hughes Medical Institute.

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Author notes

  1. Rosa Nguyen and Anand G. Patel have contributed equally.

Authors and Affiliations

  1. Department of Oncology, St. Jude Children’s Research Hospital, Memphis, TN, USA

    Rosa Nguyen, Anand G. Patel, Elizabeth A. Stewart & Wayne L. Furman

  2. Pediatric Oncology Branch, National Cancer Institute, Bethesda, MD, USA

    Rosa Nguyen

  3. Department of Developmental Neurobiology, St. Jude Children’s Research Hospital, Memphis, TN, USA

    Anand G. Patel, Lyra M. Griffiths, Jason Dapper, Elizabeth A. Stewart, Jim Houston & Michael A. Dyer

  4. Center for In Vivo Imaging and Therapeutics (CIVIT), St. Jude Children’s Research Hospital, Memphis, TN, USA

    Melissa Johnson & Walter J. Akers

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  1. Rosa Nguyen
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Contributions

RN designed and conducted all experiments, analyzed and interpreted all data, and drafted and reviewed the manuscript; AGP analyzed and interpreted the scRNA-seq data and reviewed the manuscript; LMG maintained the MISTRG colony and supplied the mice and reviewed the manuscript; EAS maintained the MISTRG colony, conducted murine experiments requested in revisions, reviewed the manuscript; JD maintained the MISTRG colony, conducted murine experiments requested in revisions, reviewed the manuscript; JH performed flow cytometry analysis and reviewed the manuscript; MJ and WJA performed tumor injections and ultrasound-based tumor measurements and reviewed the manuscript; WLF: provided patient samples and clinical data and reviewed the manuscript; MAD generated the hypothesis, designed, funded, supervised the study, interpreted data, and reviewed and edited the draft.

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Correspondence to Rosa Nguyen.

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Nguyen, R., Patel, A.G., Griffiths, L.M. et al. Next-generation humanized patient-derived xenograft mouse model for pre-clinical antibody studies in neuroblastoma. Cancer Immunol Immunother 70, 721–732 (2021). https://doi.org/10.1007/s00262-020-02713-6

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Keywords

  • Pediatric oncology
  • Natural killer cells
  • Neuroblastoma
  • Antibody therapy