Neuroblastoma is the most common extracranial solid tumor in children and originates from poorly differentiated neural crest progenitors. High-risk neuroblastoma patients frequently present with metastatic disease at diagnosis. Despite intensive treatment, patients often develop refractory disease characterized by poorly differentiated, therapy resistant cells. Although adjuvant therapy using retinoic acid (RA)-induced differentiation may increase event-free survival, in the majority of cases response to RA-therapy is inadequate. Consequently, current research aims to identify novel therapeutic targets that enhance the sensitivity to RA and induce neuroblastoma cell differentiation. The similarities between neural crest development and neuroblastoma progression provide an appealing starting point. During neural crest development the EMT-transcription factor SNAI2 plays an important role in neural crest specification as well as neural crest cell migration and survival. Here, we report that CRISPR/Cas9 mediated deletion as well as shRNA mediated knockdown of the EMT-transcription factor SNAI2 promotes cellular differentiation in a variety of neuroblastoma models. By comparing mRNA expression data from independent patient cohorts, we show that a SNAI2 activity-based gene expression signature significantly correlates with event-free survival. Loss of SNAI2 function reduces self-renewal, 3D invasion as well as metastatic spread in vivo, while strongly sensitizing neuroblastoma cells to RA-induced growth inhibition. Together, our data demonstrate that SNAI2 maintains progenitor-like features in neuroblastoma cells while interfering with RA-induced growth inhibition. We propose that targeting gene regulatory circuits, such as those controlling SNAI2 function, may allow reversion of RA-therapy resistant neuroblastoma cells to a more differentiated and therapy responsive phenotype.

中文翻译：

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转录阻遏物SNAI2损害神经母细胞瘤分化并抑制对视黄酸治疗的反应。

神经母细胞瘤是儿童中最常见的颅外实体瘤，起源于分化差的神经rest祖细胞。高危神经母细胞瘤患者在诊断时经常出现转移性疾病。尽管进行了强化治疗，但患者通常仍会以难治性疾病为特征，这些疾病的特点是分化差的治疗耐药性细胞。尽管使用视黄酸（RA）诱导的分化进行辅助治疗可以增加无事件生存期，但在大多数情况下，对RA治疗的反应不足。因此，当前的研究旨在确定增强RA敏感性并诱导神经母细胞瘤细胞分化的新型治疗靶标。神经rest发育与神经母细胞瘤进展之间的相似性提供了一个有吸引力的起点。在神经c发育过程中，EMT转录因子SNAI2在神经rest规格以及神经rest细胞迁移和存活中起着重要作用。在这里，我们报道CRISPR / Cas9介导的缺失以及EMT转录因子SNAI2的shRNA介导的敲低促进了多种神经母细胞瘤模型中的细胞分化。通过比较来自独立患者队列的mRNA表达数据，我们表明基于SNAI2活性的基因表达特征与无事件生存率显着相关。SNAI2功能的丧失减少了自我更新，3D侵袭以及体内转移的扩散，同时使神经母细胞瘤细胞对RA诱导的生长抑制高度敏感。一起，我们的数据表明，SNAI2可以在神经母细胞瘤细胞中维持祖细胞样功能，同时干扰RA诱导的生长抑制。我们建议靶向基因调控电路，如那些控制SNAI2功能的基因调控电路，可以使RA治疗耐药的神经母细胞瘤细胞恢复为更具分化和治疗反应性的表型。