Espallergues Julie, Cadiet Julien, Souab Fouzia, Choquet Olivier, Swisser Fabien, Bigeleisen Paul, Maleysson Vincent, Sola Marie-Laure, van Hameren Gerben, Tricaud Nicolas
Nervosave Therapeutics, 371 rue du Professeur Blayac, Montpellier, France.
CHU de Nantes, Bd Jacques Monod, St Herblain, Nantes, France.
Mol Ther Methods Clin Dev. 2025 Aug 5;33(3):101548. doi: 10.1016/j.omtm.2025.101548. eCollection 2025 Sep 11.
AAV-based gene therapy represents an attractive treatment for hereditary peripheral neuropathies Charcot-Marie-Tooth diseases. We recently showed that AAV2/9 vector-expressing GFP, locally injected into mouse and rat sciatic nerves, transduced a large amount of myelinating Schwann cells (mSCs). The local delivery of a similar vector expressing a small hairpin RNA (shRNA) targeting PMP22 mRNA prevented CMT1A disease in a rat disease model. Here, we investigated the regional anesthesia standard perineural injection route in non-human primates using AAV2/9 expressing GFP and AAV2/9 expressing shRNA targeting human and monkey PMP22 mRNA. Injecting at multiple sites to cover the largest part of arm and leg nerves, we found that AAV2/9 easily crossed perineurium sheaths to transduce up to 90% of mSCs. Injections on 10 nerves in each animal did not generate any adverse effects. Injected nerves functioned properly, and fine motor dexterity remained unaffected. Enzymatic, metabolite, and blood cytometry were marginally affected, and no systemic inflammation was detected. Locally, we detected a slight to moderate perineural infiltration of monocytes only with vector-expressing GFP. Vector-expressing PMP22 shRNA decreased PMP22 expression in a dose-dependent manner and levels remained physiological. Perineural injections appear to be safe, well-tolerated, and efficient to deliver AAV2/9-based gene therapy vector to treat Charcot-Marie-Tooth diseases.
基于腺相关病毒（AAV）的基因疗法是治疗遗传性周围神经病——夏科-马里-图斯病（Charcot-Marie-Tooth diseases）的一种有吸引力的治疗方法。我们最近发现，将表达绿色荧光蛋白（GFP）的AAV2/9载体局部注射到小鼠和大鼠的坐骨神经中，可转导大量的髓鞘形成施万细胞（mSCs）。在大鼠疾病模型中，局部递送表达靶向PMP22 mRNA的小发夹RNA（shRNA）的类似载体可预防CMT1A疾病。在此，我们使用表达GFP的AAV2/9和表达靶向人类和猴PMP22 mRNA的shRNA的AAV2/9，研究了非人类灵长类动物的区域麻醉标准经神经注射途径。通过在多个部位注射以覆盖手臂和腿部神经的最大部分，我们发现AAV2/9很容易穿过神经束膜鞘，转导高达90%的mSCs。在每只动物的10条神经上进行注射未产生任何不良反应。注射后的神经功能正常，精细运动灵活性未受影响。酶、代谢物和血细胞计数仅受到轻微影响，未检测到全身炎症。在局部，仅在表达GFP载体的情况下，我们检测到单核细胞有轻微至中度的神经束膜浸润。表达PMP22 shRNA的载体以剂量依赖的方式降低了PMP22的表达，且水平保持在生理范围内。经神经注射似乎是安全的、耐受性良好的，并且能够有效地递送基于AAV2/9的基因治疗载体来治疗夏科-马里-图斯病。
