Bioluminescence-optogenetics-mediated gene therapy in a sleep-disordered breathing mouse model

• Published in: Biomedicine & pharmacotherapy 2024-09, Vol.178, p.117159, Article 117159
• Main Authors: Wang, Yixuan, Liu, Xu, Zhang, Qingfeng, Zhao, Dong, Zhou, Beini, Pan, Zhou, Zha, Shiqian, Hu, Ke
• Format: Article
• Language: English
• Subjects: Adeno-associated virus; Animals; Bioluminescence-optogenetics; Coelenterazine; Dependovirus - genetics; Disease Models, Animal; Genetic Therapy - methods; Genioglossus; Hypoglossal Nerve; Luminescent Measurements; Male; Mice; Mice, Inbred C57BL; Motor Neurons - metabolism; Obstructive sleep apnea; Optogenetics - methods; Sleep Apnea Syndromes - genetics; Sleep Apnea Syndromes - therapy
• ISSN: 0753-3322; 1950-6007; 1950-6007
• DOI: 10.1016/j.biopha.2024.117159

Obstructive sleep apnea (OSA) incurs a huge individual, societal, and economic burden. Specific and selective targeting of hypoglossal motor neurons could be an effective means to treat OSA. Bioluminescent-optogenetics (BL-OG) is a novel genetic regulatory approach in which luminopsins, fusion proteins of light-generating luciferase and light-sensing ion channels, increase neuronal excitability when exposed to a suitable substrate. Here we develop and validate the feasibility of BL-OG for sleep-disordered breathing (SDB). Upon confirming that diet-induced obese mice represent an excellent SDB model, we employed a method of targeting the hypoglossal nucleus (12 N) by peripherally injecting retrogradely transported rAAV2/Retro. With AAV transduction, the eLMO3 protein is expressed in hypoglossal motor neurons (HMN); administration of CTZ results in production of bioluminescence that in turn activates the tethered channelrhodopsin, leading to an increase in the firing of HMN and a 2.7 ± 0.8-fold increase in phasic activity of the genioglossus muscle, a 7.6 ± 1.8-fold increase in tonic activity, and improvements in hypoventilation and apnea index without impacting sleep structure. This is therefore the first study to leverage the rAAV2/Retro vector to execute the BL-OG approach in SDB, which amplified genioglossus muscle discharge activity and increased airflow in mice after activation. This study marks the pioneering utilization of BL-OG in SDB research. [Display omitted] •Diet-induced obese (DIO) mice represent an excellent model of SDB.•Compared to rAAV2/9 and rAAV2/11, the rAAV2/Retro serotype has a higher retrograde infection efficiency.•The BL-OG approach, which was used for the first time in SDB research, has demonstrated a positive safety profile.•In virus-infected mice, CTZ administration increases muscle activity, improves hypoventilation and AI, without altering sleep structure.