Abou Haidar Edwina, Prabhakar Shilpa, Cheah Pike See, Hanlon Killian S, Espinoza Paula, Crain Adam V, Patel Nikita, Radcliff Greta W, Cheng Ming, Hernández Iván Coto, Minderler Steven, de la Cruz Demitri, Ng Carrie, da Hora Cintia Carla, Charest Alain, Stemmer-Rachamimov Anat, Jowett Nate, Breakefield Xandra O, Maguire Casey A
Molecular Neurogenetics Unit, Department of Neurology, The Massachusetts General Hospital, Charlestown MA and Program in Neuroscience, Harvard Medical School, Boston, MA, USA.
Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia.
Gene Ther. 2025 Jun 10. doi: 10.1038/s41434-025-00542-9.
Genetic diseases such as Neurofibromatosis type 1 (NF1) and Charcot-Marie Tooth disease involve Schwann cells (SCs) associated with peripheral nerves. Gene therapy using adeno-associated virus (AAV) vector mediated gene delivery is a promising strategy to treat these diseases. However, AAV-mediated transduction of SCs in vivo after intravascular delivery is relatively inefficient, with a lack of extensive characterization of different capsids to date. Here, we performed an in vivo selection with an AAV9 capsid peptide display library in a mouse model of NF1. We chose one capsid variant, AAV-SC3, which was present in NF1 nerves for comparison to two benchmark capsids after systemic injection. AAV-SC3 significantly outperformed one of the two benchmark capsids at levels of transgene mRNA in the neurofibroma. Immunofluorescence microscopy revealed transgene expressing Sox10-positive SCs throughout the neurofibroma with AAV-SC3 injection. Next, we performed a pooled screen with four of the top capsids from our initial selection and AAV9 and identified one capsid, AAV-SC4, with enhanced biodistribution to and transduction of normal sciatic nerve in mice. This capsid displayed a peptide with a known laminin-binding motif, which may provide a conduit for future laminin-targeting strategies. Our results provide a baseline for future AAV-based gene therapies developed for NF1 or other diseases that affect SCs.
诸如1型神经纤维瘤病（NF1）和夏科-马里-图思病等遗传性疾病涉及与周围神经相关的施万细胞（SCs）。使用腺相关病毒（AAV）载体介导的基因递送进行基因治疗是治疗这些疾病的一种有前景的策略。然而，血管内递送后AAV介导的SCs体内转导效率相对较低，且迄今为止缺乏对不同衣壳的广泛表征。在此，我们在NF1小鼠模型中使用AAV9衣壳肽展示文库进行了体内筛选。我们选择了一种衣壳变体AAV-SC3，其存在于NF1神经中，以便在全身注射后与两种基准衣壳进行比较。在神经纤维瘤中的转基因mRNA水平上，AAV-SC3显著优于两种基准衣壳之一。免疫荧光显微镜显示，注射AAV-SC3后，整个神经纤维瘤中均有表达转基因的Sox10阳性SCs。接下来，我们用最初筛选出的四种顶级衣壳和AAV9进行了混合筛选，确定了一种衣壳AAV-SC4，其对小鼠正常坐骨神经的生物分布和转导能力增强。这种衣壳展示了一种具有已知层粘连蛋白结合基序的肽，这可能为未来的层粘连蛋白靶向策略提供一条途径。我们的结果为未来针对NF1或其他影响SCs的疾病开发的基于AAV的基因治疗提供了一个基线。
