Neuroblastoma (NB), the most prevalent extracranial solid tumor in children, poses a significant therapeutic challenge due to its metastasis and high heterogeneity. A recent study leveraging single-cell RNA sequencing (scRNA-seq) has uncovered vital molecular mechanisms underlying NB's progression and metastasis, shedding light on potential therapeutic targets. The research analyzed primary tumors and matched metastases from NB patients, revealing a 'starter' subpopulation of tumor cells responsible for initiating metastasis. By applying various analytical approaches, including evolutionary trajectory analysis and cell-state differentiation prediction, the study delineated the transcriptional landscape of NB and identified a signature associated with poor prognosis.

The investigation involved collecting tumor samples from six NB patients, encompassing both primary and metastatic sites. These samples were subjected to scRNA-seq to generate a comprehensive single-cell expression atlas. The analysis revealed 11 distinct clusters, each with unique transcriptional features, highlighting the heterogeneity of NB. Through inferred copy number variation (CNV) analysis and whole genomic sequencing (WGS), malignant cells were distinguished from non-malignant cells, facilitating a deeper understanding of tumor cell populations.

The study's trajectory analysis pinpointed a specific cluster, termed 'starter' cells, as the likely initiators of NB's metastatic spread. These cells, identified in both primary tumors and metastases, exhibited a high proliferative capacity and were characterized by an active cell cycle and DNA repair pathways. The 'starter' cells also demonstrated partial epithelial-to-mesenchymal transition (EMT), a process associated with enhanced migratory and invasive properties of cancer cells, suggesting a role in NB's metastatic spread.

To further probe the interactions between NB cells and their microenvironment, the researchers analyzed scRNA-seq data from non-malignant cells within the tumor ecosystem. They discovered robust interactions, particularly mediated by the TGFβ signaling pathway, between the 'starter' cells and various immune and stromal cells. This finding points to a complex crosstalk that may influence tumor progression and immune evasion.

In a significant discovery, the study identified a 150-gene 'starter' cell signature that was associated with poor clinical outcomes across multiple datasets. The signature's predictive power for unfavorable prognosis was robust across different sequencing platforms and patient cohorts, underscoring its potential utility in clinical settings.

Despite the study's comprehensive approach, it acknowledged certain limitations, including the relatively small sample size due to the challenges of collecting paired primary and metastatic NB samples. However, the findings were validated using public datasets and independent experimental approaches, bolstering the study's conclusions.

In conclusion, this study provides a detailed molecular roadmap of NB's metastatic journey, identifying a key cell subpopulation and associated gene signature that could inform future therapeutic strategies. The insights into the cellular and molecular dynamics of NB have the potential to improve prognostication and guide targeted interventions for this aggressive pediatric cancer.

DOI: 10.1007/s11684-024-1081-7