Upton Dannielle H, Liu Jie, George Sandra M, Valvi Santosh, Ung Caitlin, Rayner Benjamin S, Gopalakrishnan Anjana, Pandher Ruby, Khan Aaminah, Venkat Pooja, Mayoh Chelsea, Holliday Holly, Yeung Nicole, Nguyen Hieu, Franshaw Laura, Ehteda Anahid, Shen Han, Farruggia Giovanna, Orienti Isabella, Reynolds C Patrick, Tsoli Maria, Ziegler David S
School of Clinical Medicine, University of New South Wales, Sydney, NSW, Australia.
Children's Cancer Institute, Lowy Cancer Research Centre, University of New South Wales, Sydney, NSW, Australia.
Neuro Oncol. 2025 Sep 8;27(7):1813-1828. doi: 10.1093/neuonc/noaf035.
Diffuse midline glioma, characterized by H3K27 alteration (DMG), is the predominant high-grade glioma in children. It commonly originates in the brainstem, yet effective treatments for these patients remain elusive.
To identify novel therapies for DMG, we conducted high-throughput drug screens (HTS) using biologically active, clinically approved compounds against DMG neurospheres. Multiple primary DMG cultures were utilized in conjunction with in vitro cytotoxicity and clonogenic assays to validate the efficacy of top compounds. Molecularly diverse patient-derived and transgenic DMG orthotopic models were employed to assess therapeutic efficacy alongside pharmacokinetic and immunohistochemical analyses. Mechanistic studies, including RNA sequencing, western blotting, and flow cytometry, were conducted to elucidate the antitumor efficacy of the most promising compound, fenretinide, in DMG cells.
Through HTS, 6 compounds were identified and validated for their potent cytotoxic activity. However, most of these compounds failed to improve survival in an orthotopic Diffuse Midline Glioma (DMG) model due to limited blood-brain barrier (BBB) penetration. In contrast, fenretinide exhibited effective BBB penetration, significantly enhancing the survival of tumor-bearing animals. Mechanistic studies revealed that fenretinide increased reactive oxygen species (ROS) generation and induced apoptosis by inhibiting PDGFRα. RNA-sequencing further elucidated that fenretinide upregulates the Unfolded Protein Response (UPR) and endoplasmic reticulum (ER) stress pathways while downregulating neurogenesis. The in vivo antitumor efficacy of 2 fenretinide formulations was demonstrated in PDGFRα-amplified and transgenic DMG models.
This comprehensive study has identified new DMG therapeutic vulnerabilities and highlights fenretinide as a brain-penetrant, anti-DMG agent.
以H3K27改变为特征的弥漫性中线胶质瘤（DMG）是儿童中主要的高级别胶质瘤。它通常起源于脑干，但针对这些患者的有效治疗方法仍然难以捉摸。
为了确定DMG的新疗法，我们使用具有生物活性、临床批准的化合物对DMG神经球进行了高通量药物筛选（HTS）。多个原发性DMG培养物与体外细胞毒性和克隆形成试验一起用于验证顶级化合物的疗效。利用分子多样性的患者来源和转基因DMG原位模型评估治疗效果，并进行药代动力学和免疫组织化学分析。进行了包括RNA测序、蛋白质印迹和流式细胞术在内的机制研究，以阐明最有前景的化合物芬维A胺在DMG细胞中的抗肿瘤疗效。
通过HTS，鉴定并验证了6种具有强细胞毒性活性的化合物。然而，由于血脑屏障（BBB）穿透有限，这些化合物中的大多数未能改善原位弥漫性中线胶质瘤（DMG）模型中的生存率。相比之下，芬维A胺表现出有效的BBB穿透性，显著提高了荷瘤动物的生存率。机制研究表明，芬维A胺通过抑制PDGFRα增加活性氧（ROS）生成并诱导凋亡。RNA测序进一步阐明，芬维A胺上调未折叠蛋白反应（UPR）和内质网（ER）应激途径，同时下调神经发生。在PDGFRα扩增和转基因DMG模型中证明了两种芬维A胺制剂的体内抗肿瘤疗效。
这项全面的研究确定了新的DMG治疗弱点，并突出了芬维A胺作为一种可穿透血脑屏障的抗DMG药物。
