Zhou Haibin, Dai Jiajing, Li Dong, Wang Luyao, Ye Meng, Hu Xiaoling, LoTurco Joseph, Hu Ji, Sun Wenzhi
School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China; Chinese Institute for Brain Research, Beijing 102206, China.
School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China.
Mol Ther. 2025 Mar 5;33(3):1166-1179. doi: 10.1016/j.ymthe.2025.01.006. Epub 2025 Jan 10.
The development of efficient and targeted methods for delivering DNA in vivo has long been a major focus of research. In this study, we introduce a gene delivery approach admitted by small metabolites (gDAM) for the efficient and targeted delivery of naked DNA into astrocytes in the adult brains of mice. gDAM uses a straightforward combination of DNA and small metabolites, including glycine, L-proline, L-serine, L-histidine, D-alanine, Gly-Gly, and Gly-Gly-Gly, to achieve astrocyte-specific delivery of naked DNA, resulting in transient and robust gene expression in these cells. Using gDAM, we successfully co-deliver the PiggyBac transposon and the CRISPR-Cas9 system to induce long-term overexpression of the oncogene EGFRvIII and knockout of tumor suppressor genes Nf1, Pten, and Trp53 in astrocytes, leading to the development of astrocyte-derived gliomas in immunocompetent mice. Furthermore, gDAM facilitates the delivery of naked DNA to peripheral glioma astrocytes. The overexpression of interferon-β and granulocyte-macrophage colony-stimulating factor in these peripheral glioma astrocytes significantly prolongs the overall survival of mice bearing 73C glioma cells. This approach offers a new perspective on developing gene delivery systems that specifically target astrocytes to meet the varied needs of both research and gene therapy. The innovative strategy behind gDAM is expected to provide fresh inspiration in the quest for DNA delivery to other tissues, such as skeletal muscle and skin.
长期以来，开发高效且有针对性的体内DNA递送方法一直是研究的主要重点。在本研究中，我们引入了一种由小分子代谢物介导的基因递送方法（gDAM），用于将裸DNA高效且有针对性地递送至成年小鼠大脑中的星形胶质细胞。gDAM使用DNA与小分子代谢物（包括甘氨酸、L-脯氨酸、L-丝氨酸、L-组氨酸、D-丙氨酸、甘氨酰甘氨酸和甘氨酰甘氨酰甘氨酸）的直接组合，实现裸DNA向星形胶质细胞的特异性递送，从而在这些细胞中产生瞬时且强劲的基因表达。使用gDAM，我们成功地共递送了PiggyBac转座子和CRISPR-Cas9系统，以诱导星形胶质细胞中癌基因EGFRvIII的长期过表达以及肿瘤抑制基因Nf1、Pten和Trp53的敲除，从而在免疫活性小鼠中引发星形胶质细胞源性胶质瘤的形成。此外，gDAM有助于将裸DNA递送至外周胶质瘤星形胶质细胞。这些外周胶质瘤星形胶质细胞中干扰素-β和粒细胞巨噬细胞集落刺激因子的过表达显著延长了携带73C胶质瘤细胞的小鼠的总体生存期。这种方法为开发特异性靶向星形胶质细胞的基因递送系统提供了新的视角，以满足研究和基因治疗的各种需求。gDAM背后的创新策略有望在寻求向其他组织（如骨骼肌和皮肤）递送DNA方面提供新的灵感。
