Reflection of neuroblastoma intratumor heterogeneity in the new OHC‐NB1 disease model

Abstract

Accurate modeling of intratumor heterogeneity presents a bottleneck against drug testing. Flexibility in a preclinical platform is also desirable to support assessment of different endpoints. We established the model system, OHC‐NB1, from a bone marrow metastasis from a patient diagnosed with MYCN‐amplified neuroblastoma and performed whole‐exome sequencing on the source metastasis and the different models and passages during model development (monolayer cell line, 3D spheroid culture and subcutaneous xenograft tumors propagated in mice). OHC‐NB1 harbors a MYCN amplification in double minutes, 1p deletion, 17q gain and diploid karyotype, which persisted in all models. A total of 80–540 single‐nucleotide variants (SNVs) was detected in each sample, and comparisons between the source metastasis and models identified 34 of 80 somatic SNVs to be propagated in the models. Clonal reconstruction using the combined copy number and SNV data revealed marked clonal heterogeneity in the originating metastasis, with four clones being reflected in the model systems. The set of OHC‐NB1 models represents 43% of somatic SNVs and 23% of the cellularity in the originating metastasis with varying clonal compositions, indicating that heterogeneity is partially preserved in our model system.

Perfusion-Based Bioreactor Culture and Isothermal Microcalorimetry for Preclinical Drug Testing with the Carbonic Anhydrase Inhibitor SLC-0111 in Patient-Derived Neuroblastoma

“…ECM support can be provided, e.g., in the form of scaffolds. Tissue-derived tumor spheres (TDTSs) isolated from patient tumor cells are used in other models, which are more often derived from metastatic bone marrow aspirate cultured in Matrigel [ 48 , 49 , 50 , 51 ]. These have successfully been applied to neuroblastoma.…”

Emerging Neuroblastoma 3D In Vitro Models for Pre-Clinical Assessments

“…Screening for additional antigens specific for neuroblastoma is, however, necessary for more accurate selection of tumor cells with stem features that are often responsible for drug resistance development and disease recurrence. Another TDTS model resembling neuroblastoma intratumoral heterogeneity has been reported by Thole and colleagues (30). The primary neuroblastoma TDTSs culture derived from a bone marrow aspirate with 80% tumor cell infiltration can be cultured in Matrigel.…”

“…Neuroblastoma cells grown as 3D spheroids maintain the tumorigenic capability in a xenotransplantation mouse model through five passages. Importantly, these TDTSs model systems partly reflect the genetic and clonal heterogeneity of the initial biopsy (30). Altogether, the reported neuroblastoma TDTSs represent essential initial steps toward more sophisticated 3D neuroblastoma modeling suitable for pre-clinical testing.…”

“…TDTSs are obtained by tumor tissue mechanical dissociation (26). Due to the origin of primary cells, this in vitro model system more closely reflects the genetic and clonal heterogeneity of the native tumor, thus providing a more accurate pre-clinical platform (30). Despite the fact that these tailored models can lead to an improved level of prediction, their development and application are still a challenge as sample collection and size, protocol standardization and data reproducibility are critical issues for neuroblastoma.…”

“…Despite the fact that these tailored models can lead to an improved level of prediction, their development and application are still a challenge as sample collection and size, protocol standardization and data reproducibility are critical issues for neuroblastoma. More easily established are neuroblastomaderived spheroids generated from the bone marrow aspirates of patients diagnosed with stage 4 metastatic disease (30). In some cases, surgically removed tumor resections also allow in vitro reproduction of neuroblastoma.…”