Nieland Lisa, Vrijmoet Anne B, Jetten Isabelle W, Rufino-Ramos David, de Reus Alexandra J E M, Breyne Koen, Kleinstiver Benjamin P, Maguire Casey A, Broekman Marike L D, Breakefield Xandra O, Abels Erik R
Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA; Department of Cell and Chemical Biology, Leiden University Medical Center, 2300 RC Leiden, the Netherlands.
Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA.
Mol Ther. 2025 Jan 8;33(1):133-151. doi: 10.1016/j.ymthe.2024.11.023. Epub 2024 Nov 19.
Glioblastoma (GB), the most aggressive tumor of the central nervous system (CNS), has poor patient outcomes with limited effective treatments available. MicroRNA-21 (miR-21(a)) is a known oncogene, abundantly expressed in many cancer types. miR-21(a) promotes GB progression, and lack of miR-21(a) reduces the tumorigenic potential. Here, we propose a single adeno-associated virus (AAV) vector strategy targeting mmu-miR-21a using the Staphylococcus aureus Cas9 ortholog (SaCas9) guided by a single-guide RNA (sgRNA). Our results demonstrate that AAV8 is a well-suited AAV serotype to express SaCas9-KKH/sgRNA at the tumor site in an orthotopic GB model. The SaCas9-KKH induced a genomic deletion, resulting in lowered mmu-miR-21a levels in the brain, leading to reduced tumor growth and improved overall survival. In this study, we demonstrated that disruption of genomic mmu-miR-21a with a single AAV vector influenced glioma development, resulting in beneficial anti-tumor outcomes in GB-bearing mice.
胶质母细胞瘤（GB）是中枢神经系统（CNS）中最具侵袭性的肿瘤，患者预后较差，有效治疗方法有限。微小RNA-21（miR-21(a)）是一种已知的致癌基因，在许多癌症类型中大量表达。miR-21(a)促进GB进展，而缺乏miR-21(a)会降低致瘤潜力。在此，我们提出一种单一腺相关病毒（AAV）载体策略，使用由单向导RNA（sgRNA）引导的金黄色葡萄球菌Cas9直系同源物（SaCas9）靶向小鼠miR-21a。我们的结果表明，在原位GB模型中，AAV8是一种非常适合在肿瘤部位表达SaCas9-KKH/sgRNA的AAV血清型。SaCas9-KKH诱导基因组缺失，导致大脑中mmu-miR-21a水平降低，从而减少肿瘤生长并改善总体生存率。在本研究中，我们证明用单一AAV载体破坏基因组mmu-miR-21a会影响胶质瘤发展，在荷GB小鼠中产生有益的抗肿瘤结果。
