Targeting N-Cadherin and Tubulin α 1A in Neuroblastoma Bone Marrow Metastasis: Insights from Single-Cell Analysis and Drug Screening

Neuroblastoma (NB) with metastasis to bone marrow (BM) usually results in dismal survival. The mechanisms underlying BM metastasis remain largely unclear. In this study, single-cell transcriptome of NB with and without BM metastasis were analyzed, and 146 marker genes of NB cells were identified. Using two-sample Mendelian randomization, N-Cadherin (CDH2) and tubulin α 1A (TUBA1A) were identified as key genes possibly affecting BM metastasis. Functional investigations suggest that CDH2 regulates cell cycle and DNA replication, whereas TUBA1A impacts cell adhesion molecules and the cAMP signaling pathway. Dysregulated expression of CDH2 or TUBA1A also alters the immune microenvironment by immune cells and chemokines. Subsequent experiments validated that both CDH2 and TUBA1A were up-regulated in NB cell lines and high-risk patient samples, compared with low-risk cases. Knockdown of CDH2 or TUBA1A led to cell cycle arrest and significantly inhibited NB cell proliferation and migration. Moreover, an oxidation-reduction cycling agent, 2,3-dimethoxy-1,4-naphthoquinone, was identified as a candidate compound with inhibitory activity on NB cells by diminishing CDH2 or TUBA1A expression levels. Together, these results highlight CDH2 and TUBA1A as novel therapeutic targets for NB with BM metastases. The efficacy of 2,3-dimethoxy-1,4-naphthoquinone in suppressing NB cell growth and CDH2/TUBA1A expression suggests its potential for clinical application.