Background Long non-coding RNAs (lncRNAs) exhibit highly cell type-specific expression and function, making this class of transcript attractive for targeted cancer therapy. However, the vast majority of lncRNAs have not been tested as potential therapeutic targets, particularly in the context of currently used cancer treatments. Malignant glioma is rapidly fatal, and ionizing radiation is part of the current standard-of-care used to slow tumor growth in both adult and pediatric patients. Results We use CRISPR interference (CRISPRi) to screen 5689 lncRNA loci in human glioblastoma (GBM) cells, identifying 467 hits that modify cell growth in the presence of clinically relevant doses of fractionated radiation. Thirty-three of these lncRNA hits sensitize cells to radiation, and based on their expression in adult and pediatric gliomas, nine of these hits are prioritized as lncRNA Glioma Radiation Sensitizers ( lncGRS ). Knockdown of lncGRS-1 , a primate-conserved, nuclear-enriched lncRNA, inhibits the growth and proliferation of primary adult and pediatric glioma cells, but not the viability of normal brain cells. Using human brain organoids comprised of mature neural cell types as a three-dimensional tissue substrate to model the invasive growth of glioma, we find that antisense oligonucleotides targeting lncGRS-1 selectively decrease tumor growth and sensitize glioma cells to radiation therapy. Conclusions These studies identify lncGRS-1 as a glioma-specific therapeutic target and establish a generalizable approach to rapidly identify novel therapeutic targets in the vast non-coding genome to enhance radiation therapy.

中文翻译：

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基于 CRISPRI 的辐射调节剂筛选可识别胶质瘤中的长非编码 RNA 治疗靶点

背景 长链非编码 RNA (lncRNAs) 表现出高度的细胞类型特异性表达和功能，使此类转录物对靶向癌症治疗具有吸引力。然而，绝大多数 lncRNA 尚未作为潜在的治疗靶点进行测试，特别是在目前使用的癌症治疗的背景下。恶性胶质瘤会迅速致命，而电离辐射是目前用于减缓成人和儿童患者肿瘤生长的标准护理的一部分。结果我们使用 CRISPR 干扰 (CRISPRi) 来筛选人类胶质母细胞瘤 (GBM) 细胞中的 5689 个 lncRNA 基因座，确定在临床相关剂量的分次辐射存在下改变细胞生长的 467 个命中。其中 33 个 lncRNA 命中使细胞对辐射敏感，并基于它们在成人和儿童神经胶质瘤中的表达，这些命中中的九个被优先考虑为 lncRNA 胶质瘤辐射敏化剂 ( lncGRS )。敲低 lncGRS-1（一种灵长类动物保守的、富含核的 lncRNA）可抑制原代成人和小儿神经胶质瘤细胞的生长和增殖，但不会抑制正常脑细胞的活力。使用由成熟神经细胞类型组成的人脑类器官作为三维组织基质来模拟神经胶质瘤的侵袭性生长，我们发现靶向 lncGRS-1 的反义寡核苷酸选择性地减少肿瘤生长并使神经胶质瘤细胞对放射治疗敏感。结论 这些研究将 lncGRS-1 确定为神经胶质瘤特异性治疗靶点，并建立了一种通用方法来快速确定大量非编码基因组中的新治疗靶点，以增强放射治疗。