High-risk neuroblastoma is often distinguished by amplification of MYCN and loss of differentiation potential. We performed high-throughput drug screening of epigenetic-targeted therapies across a large and diverse tumor cell line panel and uncovered the hypersensitivity of neuroblastoma cells to GSK-J4, a small-molecule dual inhibitor of lysine 27 of histone 3 (H3K27) demethylases ubiquitously transcribed tetratricopeptide repeat, X chromosome (UTX), and histone demethylase Jumonji D3 (JMJD3). Mechanistically, GSK-J4 induced neuroblastoma differentiation and endoplasmic reticulum (ER) stress, with accompanying up-regulation of p53 up-regulated modulator of apoptosis (PUMA) and induction of cell death. Retinoic acid (RA)–resistant neuroblastoma cells were sensitive to GSK-J4. In addition, GSK-J4 was effective at blocking the growth of chemorefractory and patient-derived xenograft models of high-risk neuroblastoma in vivo. Furthermore, GSK-J4 and RA combination increased differentiation and ER stress over GSK-J4 effects and limited the growth of neuroblastomas resistant to either drug alone. In MYCN-amplified neuroblastoma, PUMA induction by GSK-J4 sensitized tumors to the B cell lymphoma 2 (BCL-2) inhibitor venetoclax, demonstrating that epigenetic-targeted therapies and BCL-2 homology domain 3 mimetics can be rationally combined to treat this high-risk subset of neuroblastoma. Therefore, H3K27 demethylation inhibition is a promising therapeutic target to treat high-risk neuroblastoma, and H3K27 demethylation can be part of rational combination therapies to induce robust antineuroblastoma activity.

高危神经母细胞瘤常通过MYCN扩增来区分和分化潜能的丧失。我们在一个庞大而多样的肿瘤细胞系中对表观遗传靶向疗法进行了高通量药物筛选，并发现神经母细胞瘤细胞对 GSK-J4 的超敏反应，GSK-J4 是一种普遍存在的组蛋白 3 (H3K27) 去甲基化酶的赖氨酸 27 的小分子双重抑制剂转录的四肽重复序列、X 染色体 (UTX) 和组蛋白去甲基化酶 Jumonji D3 (JMJD3)。从机制上讲，GSK-J4 诱导神经母细胞瘤分化和内质网 (ER) 应激，伴随 p53 上调凋亡调节剂 (PUMA) 的上调和细胞死亡的诱导。视黄酸 (RA) 抗性神经母细胞瘤细胞对 GSK-J4 敏感。此外，GSK-J4 可有效阻断体内高危神经母细胞瘤的化学难治性和患者来源的异种移植模型的生长。此外，与 GSK-J4 作用相比，GSK-J4 和 RA 组合增加了分化和内质网应激，并限制了对单独药物耐药的神经母细胞瘤的生长。在MYCN扩增的神经母细胞瘤，GSK-J4 诱导的 PUMA 使肿瘤对 B 细胞淋巴瘤 2 (BCL-2) 抑制剂 venetoclax 敏感，表明表观遗传靶向疗法和 BCL-2 同源域 3 模拟物可以合理组合来治疗这种高神经母细胞瘤的风险子集。因此，H3K27 去甲基化抑制是治疗高危神经母细胞瘤的一个有前景的治疗靶点，并且 H3K27 去甲基化可以作为合理组合疗法的一部分，以诱导强大的抗神经母细胞瘤活性。