Knapman Fiona L, Cohen E Myfanwy, Kulaga Tom, Lovell Nigel H, Lisowski Leszek, Burke Peter G R, Bilston Lynne E
Neuroscience Research Australia, Sydney, NSW 2031, Australia.
Graduate School of Biomedical Engineering, UNSW, Sydney, NSW 2033, Australia.
Mol Ther Methods Clin Dev. 2025 Aug 12;33(3):101559. doi: 10.1016/j.omtm.2025.101559. eCollection 2025 Sep 11.
Optogenetics offers a minimally invasive, low-fatigue, and temporally precise alternative to electrical stimulation for skeletal muscle control. After opsin expression in muscle cells, contraction can be stimulated with light. Obstructive sleep apnea, characterized by repeated airway collapse during sleep, suits this approach, as upper airway muscles are readily accessible via the oral cavity, and require stimulation synchronized to respiration. This study compared wild-type (adeno-associated virus 9 [AAV9]) and engineered (AAVMYO) viral vectors for the delivery of identical muscle-specific optogenetic constructs in rats. Three weeks after intramuscular injections, both vectors produced comparable opsin expression in the tongue ( = 0.54) and near-zero expression in non-target tissues. AAVMYO-treated animals had greater light-evoked increases in muscle activation than those treated with AAV9 (8.5-fold vs. 2.0-fold; < 0.0001). Conversely, AAV9-treated animals had greater light-evoked airway dilation (2.1 mm vs. 0.3 mm; = 0.02). By 12 weeks, opsin expression declined to near-zero (vs. 3 weeks; < 0.0001), light stimulation no longer increased muscle activation ( > 0.05), and anti-AAV antibodies had significantly increased ( < 0.0001). Unlike in mice, AAVMYO did not consistently outperform AAV9 in delivering gene therapy to rat muscles. Despite this, these data support optogenetics for obstructive sleep apnea, although sustained efficacy requires a better understanding of host immune responses and potentially transient immune suppression.
光遗传学为骨骼肌控制提供了一种微创、低疲劳且时间精确的电刺激替代方法。在肌肉细胞中表达视蛋白后，可用光刺激收缩。阻塞性睡眠呼吸暂停的特征是睡眠期间气道反复塌陷，适合这种方法，因为上呼吸道肌肉可通过口腔轻松触及，且需要与呼吸同步的刺激。本研究比较了野生型（腺相关病毒9 [AAV9]）和工程化（AAVMYO）病毒载体在大鼠中递送相同肌肉特异性光遗传学构建体的情况。肌肉注射三周后，两种载体在舌部产生的视蛋白表达相当（ = 0.54），在非靶组织中的表达接近零。与用AAV9处理的动物相比，用AAVMYO处理的动物光诱发的肌肉激活增加更大（8.5倍对2.0倍； < 0.0001）。相反，用AAV9处理的动物光诱发的气道扩张更大（2.1毫米对0.3毫米； = 0.02）。到12周时，视蛋白表达降至接近零（与3周相比； < 0.0001），光刺激不再增加肌肉激活（ > 0.05），且抗AAV抗体显著增加（ < 0.0001）。与小鼠不同，在将基因疗法递送至大鼠肌肉方面，AAVMYO并不始终优于AAV9。尽管如此，这些数据支持光遗传学用于阻塞性睡眠呼吸暂停，尽管持续疗效需要更好地了解宿主免疫反应并可能进行短暂的免疫抑制。
