Yang Dong, Wu Xiaoqing, Yao Yinan, Duan Mengsi, Wang Xing, Li Guoling, Guo Aiguo, Wu Meixian, Liu Yuanhua, Zheng Jin, Zhang Renxia, Li Tong, Luk Alvin, Yao Xuan, Shi Linyu, Xu Chunlong, Yang Hui
HuidaGene Therapeutics Inc., Shanghai, China.
Institute of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China.
Nat Neurosci. 2025 Jan;28(1):72-83. doi: 10.1038/s41593-024-01838-6. Epub 2024 Dec 12.
Duplication of methyl-CpG-binding protein 2 (MECP2) gene causes MECP2 duplication syndrome (MDS). To normalize the duplicated MECP2 in MDS, we developed a high-fidelity Cas13Y (hfCas13Y) system capable of targeting the MECP2 (hfCas13Y-gMECP2) messenger RNA for degradation and reducing protein levels in the brain of humanized MECP2 transgenic mice. Moreover, the intracerebroventricular adeno-associated virus (AAV) delivery of hfCas13Y-gMECP2 in newborn or adult MDS mice restored dysregulated gene expression and improved behavior deficits. Notably, treatment with AAV9-hfCas13Y-gMECP2 extended the median survival of MECP2 transgenic mice from 156.5 to 226 d. Furthermore, studies with monkeys showed a single injection of AAV9-hfCas13Y-gMECP2 was sufficient to drive robust expression of hfCas13Y in widespread brain regions, with MECP2 knockdown efficiency reaching 52.19 ± 0.03% and significantly decreased expression of biomarker gene GDF11. Our results demonstrate that the RNA-targeting hfCas13Y-gMECP2 system is an effective intervention for MDS, providing a potential strategy for treating other dosage-sensitive diseases.
甲基-CpG结合蛋白2（MECP2）基因重复会导致MECP2重复综合征（MDS）。为了使MDS中重复的MECP2正常化，我们开发了一种高保真Cas13Y（hfCas13Y）系统，该系统能够靶向MECP2（hfCas13Y-gMECP2）信使RNA进行降解，并降低人源化MECP2转基因小鼠大脑中的蛋白质水平。此外，在新生或成年MDS小鼠中通过脑室内注射腺相关病毒（AAV）递送hfCas13Y-gMECP2可恢复失调的基因表达并改善行为缺陷。值得注意的是，用AAV9-hfCas13Y-gMECP2治疗可将MECP2转基因小鼠的中位生存期从156.5天延长至226天。此外，对猴子的研究表明，单次注射AAV9-hfCas13Y-gMECP2足以在广泛的脑区中驱动hfCas13Y的强劲表达，MECP2敲低效率达到52.19±0.03%，生物标志物基因GDF11的表达显著降低。我们的结果表明，靶向RNA的hfCas13Y-gMECP2系统是治疗MDS的有效干预措施，为治疗其他剂量敏感型疾病提供了一种潜在策略。
