Lin Yu-Han, Jiang Jyun-Hong, Chuang Hui-Ching, Huang Chao-Cheng, Hsu Wen-Ming, Wu Min-Tsui, Chen Ting-Ya, Lian Wei-Shiung, Chuang Jiin-Haur
Center for Mitochondrial Research and Medicine, College of Medicine Chang Gung University, Kaohsiung Chang Gung Memorial Hospital, Kaohsiung, Taiwan.
Department of Pediatric Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung, Taiwan.
FASEB J. 2025 Apr 15;39(7):e70521. doi: 10.1096/fj.202500403R.
Neuroblastoma (NB) is a pediatric malignancy from the neural crest, where differentiation plays a key role in prognosis. We investigated cysteine and glycine-rich protein 1 (CSRP1) as a therapeutic target for NB, as it has been linked to differentiation and carcinogenesis in various cancers. Immunohistochemical analysis of archived NB samples showed a significant correlation between CSRP1 expression and differentiation. Ectopic CSRP1 expression in MYCN-amplified BE(2)-M17 cells increased sensitivity to cisplatin, promoted neurite extension, and enhanced differentiation, apoptosis, and chemosensitivity to 13cisRA. Synergistic apoptotic effects were observed with 5-aza-2'-deoxycytidine (DAC) and Poly(I:C) treatments in SK-N-AS cells implanted in xenografts, linked to upregulation of CSRP1, innate immune receptor RIG-I, and caspase-9 activation. CSRP1 expression was significantly higher in mitochondrial DNA-depleted SK-N-AS ρ0 cells, compared to parent SK-N-AS cells. Cisplatin increased CSRP1 expression further in parent cells but not in ρ0 cells. Simultaneous upregulation of caspase-8 was found in both cell types, but increased caspase-9 only in parent cells, suggesting that both intrinsic and extrinsic apoptosis pathways are involved in CSRP1 function depending on the existence of mitochondrial DNA. These findings indicate that CSRP1 is involved in differentiation, determination of apoptosis, and possibly innate immunity in NB, which endows CSRP1 with the potential to enhance the effects of 13cisRA, DAC, and Poly(I:C) in combination therapies for NB.
神经母细胞瘤（NB）是一种源自神经嵴的儿科恶性肿瘤，其中分化在预后中起关键作用。我们研究了富含半胱氨酸和甘氨酸的蛋白1（CSRP1）作为NB的治疗靶点，因为它与各种癌症的分化和致癌作用有关。对存档的NB样本进行免疫组织化学分析显示，CSRP1表达与分化之间存在显著相关性。在MYCN扩增的BE(2)-M17细胞中异位表达CSRP1可增加对顺铂的敏感性，促进神经突延伸，并增强分化、凋亡以及对13顺式维甲酸（13cisRA）的化学敏感性。在异种移植中植入的SK-N-AS细胞中，5-氮杂-2'-脱氧胞苷（DAC）和聚肌苷酸-聚胞苷酸（Poly(I:C)）处理观察到协同凋亡效应，这与CSRP1、天然免疫受体视黄酸诱导基因I（RIG-I）的上调以及半胱天冬酶-9的激活有关。与亲代SK-N-AS细胞相比，线粒体DNA缺失的SK-N-AS ρ0细胞中CSRP1表达显著更高。顺铂在亲代细胞中进一步增加CSRP1表达，但在ρ0细胞中未增加。在两种细胞类型中均发现半胱天冬酶-8同时上调，但仅在亲代细胞中半胱天冬酶-9增加，这表明根据线粒体DNA的存在情况，内源性和外源性凋亡途径均参与CSRP1功能。这些发现表明，CSRP1参与NB的分化、凋亡的决定以及可能的天然免疫，这赋予CSRP1在NB联合治疗中增强13cisRA、DAC和Poly(I:C)作用的潜力。
