Murlidharan Giridhar, Sakamoto Kensuke, Rao Lavanya, Corriher Travis, Wang Dan, Gao Guangping, Sullivan Patrick, Asokan Aravind
Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Curriculum in Genetics & Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
Mol Ther Nucleic Acids. 2016 Jul 19;5(7):e338. doi: 10.1038/mtna.2016.49.
Gene therapy using recombinant adeno-associated viral (AAV) vectors is emerging as a promising approach to treat central nervous system disorders such as Spinal muscular atrophy, Batten, Parkinson and Alzheimer disease amongst others. A critical remaining challenge for central nervous system-targeted gene therapy, silencing or gene editing is to limit potential vector dose-related toxicity in off-target cells and organs. Here, we characterize a lab-derived AAV chimeric (AAV2g9), which displays favorable central nervous system attributes derived from both parental counterparts, AAV2 and AAV9. This synthetic AAV strain displays preferential, robust, and widespread neuronal transduction within the brain and decreased glial tropism. Importantly, we observed minimal systemic leakage, decreased sequestration and gene transfer in off-target organs with AAV2g9, when administered into the cerebrospinal fluid. A single intracranial injection of AAV2g9 vectors encoding guide RNAs targeting the schizophrenia risk gene MIR137 (encoding MIR137) in CRISPR/Cas9 knockin mice resulted in brain-specific gene deletion with no detectable events in the liver. This engineered AAV vector is a promising platform for treating neurological disorders through gene therapy, silencing or editing modalities.
使用重组腺相关病毒（AAV）载体的基因治疗正在成为一种有前景的方法，用于治疗中枢神经系统疾病，如脊髓性肌萎缩症、巴顿病、帕金森病和阿尔茨海默病等。针对中枢神经系统的基因治疗、基因沉默或基因编辑，一个关键的挑战仍然是限制在非靶细胞和器官中潜在的载体剂量相关毒性。在这里，我们对一种实验室衍生的AAV嵌合体（AAV2g9）进行了表征，它表现出来自亲本AAV2和AAV9的有利的中枢神经系统特性。这种合成的AAV毒株在大脑中显示出优先、强大且广泛的神经元转导，同时减少了对神经胶质细胞的嗜性。重要的是，当将AAV2g9注入脑脊液时，我们观察到其在非靶器官中的全身泄漏、滞留和基因转移极少。在CRISPR/Cas9基因敲入小鼠中，单次颅内注射编码靶向精神分裂症风险基因MIR137（编码MIR137）的引导RNA的AAV2g9载体，导致大脑特异性基因缺失，而在肝脏中未检测到任何事件。这种工程化的AAV载体是通过基因治疗、基因沉默或编辑方式治疗神经系统疾病的一个有前景的平台。
