Comparative single-cell transcriptomics of human neuroblastoma and preclinical models reveal conservation of adrenergic cell state

Transgenic mice and organoid models, such as three-dimensional tumoroid cultures, have emerged as powerful tools for investigating cancer development and targeted therapies. Yet, the extent to which these preclinical models recapitulate the cellular identity of heterogeneous malignancies, such as neuroblastoma (NB), remains to be validated. Here we characterize the transcriptional landscape of TH-MYCN tumors by single-cell RNA sequencing (scRNA-seq) and developed novel ex vivo tumoroids. We provide a comparative analysis with murine fetal adrenal gland and human NB, to decipher the heterogeneity, cell-cell interactions, and clinical relevance of these models. Integrated analysis with fetal adrenal samples confirmed that both TH-MYCN tumors and tumoroids closely mirror the cellular profiles of normal embryonic sympathoblasts and chromaffin cells. Comprehensive comparison between tumors from NB patients and TH-MYCN mice demonstrated similarities in adrenergic tumor cell composition. Through ex vivo tumoroid cultures, we demonstrated histological resemblance, and shared transcriptional profiles with originating TH-MYCN tumors and human NB. Importantly, we identified subpopulations within tumoroids that exhibit gene expression associated with poor NB patient survival. Notably, recurrent observations of a low-proliferative chromaffin phenotype connected to the highly proliferative sympathetic phenotype may suggest that pushing sympathoblasts into a chromaffin-like state may offer an interesting therapeutic strategy for NB. Together, this study not only deepens our understanding of the widely used transgenic mouse model but also introduces a novel ex vivo model that maintains critical adrenergic cell state identity, thereby enhancing its translational potential for NB research. Tumors and ex vivo cultures from TH-MYCN transgenic mice were disaggregated and profiled using the Chromium Single Cell 3’ Library and Gel Bead Kit v2 and v3 (10X Genomics) .