Institute of Pathology and Southwest Cancer Center, Southwest Hospital, Third Military Medical University (Army Medical University) and Key Laboratory of Tumor Immunopathology, Ministry of Education of China, Chongqing, 400038, P. R. China.
Department of Clinical Laboratory, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, 400038, P. R. China.
Cancer Commun (Lond). 2022 Sep;42(9):868-886. doi: 10.1002/cac2.12334. Epub 2022 Jul 18.
Elucidation of the post-transcriptional modification has led to novel strategies to treat intractable tumors, especially glioblastoma (GBM). The ubiquitin-proteasome system (UPS) mediates a reversible, stringent and stepwise post-translational modification which is closely associated with malignant processes of GBM. To this end, developing novel therapeutic approaches to target the UPS may contribute to the treatment of this disease. This study aimed to screen the vital and aberrantly regulated component of the UPS in GBM. Based on the molecular identification, functional characterization, and mechanism investigation, we sought to elaborate a novel therapeutic strategy to target this vital factor to combat GBM.
We combined glioma datasets and human patient samples to screen and identify aberrantly regulated E3 ubiquitin ligase. Multidimensional database analysis and molecular and functional experiments in vivo and in vitro were used to evaluate the roles of HECT, UBA and WWE domain-containing E3 ubiquitin ligase 1 (HUWE1) in GBM. dCas9 synergistic activation mediator system and recombinant adeno-associated virus (rAAV) were used to endogenously overexpress full-length HUWE1 in vitro and in glioma orthotopic xenografts.
Low expression of HUWE1 was closely associated with worse prognosis of GBM patients. The ubiquitination and subsequent degradation of N-Myc mediated by HUWE1, leading to the inactivation of downstream Delta-like 1 (DLL1)-NOTCH1 signaling pathways, inhibited the proliferation, invasion, and migration of GBM cells in vitro and in vivo. A rAAV dual-vector system for packaging and delivery of dCas9-VP64 was used to augment endogenous HUWE1 expression in vivo and showed an antitumor activity in glioma orthotopic xenografts.
The E3 ubiquitin ligase HUWE1 acts through the N-Myc-DLL1-NOTCH1 signaling axis to suppress GBM progression. Antitumor activity of rAAV dual-vector delivering dCas9-HUWE1 system uncovers a promising therapeutic strategy for GBM.
对转录后修饰的阐明导致了治疗难治性肿瘤，特别是神经胶质瘤（GBM）的新策略。泛素-蛋白酶体系统（UPS）介导一种可逆的、严格的和逐步的翻译后修饰，与 GBM 的恶性过程密切相关。为此，开发针对 UPS 的新治疗方法可能有助于治疗这种疾病。本研究旨在筛选 GBM 中 UPS 的重要和异常调节成分。基于分子鉴定、功能表征和机制研究，我们试图阐述一种针对这种重要因素的新的治疗策略，以对抗 GBM。
我们结合了神经胶质瘤数据集和人类患者样本，以筛选和鉴定异常调节的 E3 泛素连接酶。多维数据库分析以及体内和体外的分子和功能实验用于评估 HECT、UBA 和 WWE 结构域包含 E3 泛素连接酶 1（HUWE1）在 GBM 中的作用。dCas9 协同激活调节剂系统和重组腺相关病毒（rAAV）用于体外和神经胶质瘤原位异种移植中过表达全长 HUWE1。
HUWE1 的低表达与 GBM 患者的预后较差密切相关。HUWE1 介导的 N-Myc 的泛素化和随后的降解，导致下游 Delta-like 1（DLL1）-NOTCH1 信号通路失活，抑制了 GBM 细胞在体外和体内的增殖、侵袭和迁移。用于包装和递送 dCas9-VP64 的 rAAV 双载体系统用于体内增强内源性 HUWE1 的表达，并显示出在神经胶质瘤原位异种移植中的抗肿瘤活性。
E3 泛素连接酶 HUWE1 通过 N-Myc-DLL1-NOTCH1 信号轴发挥作用，抑制 GBM 的进展。rAAV 双载体递送 dCas9-HUWE1 系统的抗肿瘤活性揭示了一种有前途的 GBM 治疗策略。
