High-risk neuroblastoma (HR-NB) is branded with hematogenous metastasis, relapses, and dismal long-term survival. Intensification of consolidation therapy with tandem/triple autologous stem cell (SC) rescue (with bone marrow [BM]/peripheral blood [PB] CD34⁺ selection) after myeloablative chemotherapy has improved long-term survival. However, the benefit is limited by the indication of NB cells in CD34⁺ PBSCs, CD34 expression in NB cells, and the risk of reinfusing NB cancer stem cells (NB CSCs) that could lead to post-transplant relapse. We investigated the association of CD34 surface expression (92 patients) with NB evolution/clinical outcomes. CD34 gene-level status in NB was assessed through RNA-Seq data mining (18 cohorts, n, 3324). Genetic landscape of CD34-expressing NB CSCs (CD133⁺CD34⁺) was compared with CD34⁻ CSCs (CD133⁺CD34⁻). RNA-seq data revealed equivocal association patterns of CD34 expression with patient survival. Our immunohistochemistry data revealed definite, but rare (mean, 0.73%; range 0.00–7.87%; median, 0.20%) CD34 positivity in NB. CD34⁺ significantly associated with MYCN amplification (p, 0.003), advanced disease stage (p, 0.016), and progressive disease (PD, p < 0.0009) after clinical therapy. A general high-is-worse tendency was observed in patients with relapsed disease. High CD34⁺ correlated with poor survival in patients with N-MYC-amplified HR-NB. Gene expression analysis of CD34⁺-NB CSCs identified significant up (4631) and downmodulation (4678) of genes compared with NB CSCs that lack CD34. IPA recognized the modulation of crucial signaling elements (EMT, stemness maintenance, differentiation, inflammation, clonal expansion, drug resistance, metastasis) that orchestrate NB disease evolution in CD34⁺ CSCs compared with CD34⁻ CSCs. While the function of CD34 in NB evolution requires further in-depth investigation, careful consideration should be exercised for autologous stem cell rescue with CD34⁺ selection in NB patients.

高危神经母细胞瘤 (HR-NB) 带有血行转移、复发和令人沮丧的长期生存率。清髓性化疗后强化巩固治疗与串联/三联自体干细胞 (SC) 抢救（使用骨髓 [BM]/外周血 [PB] CD34 ⁺选择）提高了长期生存率。然而，益处受限于 CD34 ⁺ PBSCs 中 NB 细胞的适应症、NB 细胞中 CD34 的表达以及可能导致移植后复发的再注入 NB 癌症干细胞 (NB CSCs) 的风险。我们调查了 CD34 表面表达（92 名患者）与 NB 进化/临床结果的关联。通过 RNA-Seq 数据挖掘评估 NB 中的 CD34 基因水平状态（18 个队列，n, 3324)。将表达 CD34 的 NB CSC (CD133 ⁺ CD34 ⁺ ) 的遗传图谱与 CD34 ^- CSC (CD133 ⁺ CD34 ^- ) 进行比较。RNA-seq 数据揭示了 CD34 表达与患者存活率的模棱两可的关联模式。我们的免疫组织化学数据显示 NB 中 CD34 阳性是确定的，但很少见（平均 0.73%；范围 0.00-7.87%；中位数，0.20%）。CD34 ⁺与临床治疗后的 MYCN 扩增 ( p , 0.003)、疾病晚期 ( p , 0.016) 和疾病进展 (PD, p  < 0.0009) 显着相关。在疾病复发的患者中观察到普遍的高是坏趋势。高CD34 ⁺与 N-MYC 扩增的 HR-NB 患者的不良生存率相关。与缺乏 CD34 的 NB CSC 相比，CD34 ⁺ -NB CSC的基因表达分析确定了基因的显着上调 (4631) 和下调 (4678)。^{与 CD34 -} CSC 相比， IPA 认识到调节 CD34 ⁺ CSC中的 NB 疾病演变的关键信号元件（EMT、干性维持、分化、炎症、克隆扩增、耐药性、转移）。虽然 CD34 在 NB 进化中的功能需要进一步深入研究，但应仔细考虑在 NB 患者中通过 CD34 ^{+选择来拯救自体干细胞。}