Gan Chunmei, Leng Mi, Liu Yu, Zheng Zhaoyue, He Siwu, Qiao Wen, Xiao Lin, Xiao Yao, Ye Jingya, Zhou Lixing, Zhou Jiao, Xiao Boduan, Zhao Wenxin, Yang Jiamei, Wu Aohan, Zhang Huiyuan, Hu Hongbo, Cen Xiaobo, Qian Zhiyong, Dong Haohao, Valencia C Alexander, Dai Lunzhi, Chow Hoi Yee, Zhang Lei, Dong Biao
National Clinical Research Center for Geriatrics and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, China.
Sichuan Real & Best Biotech Co., Ltd., Chengdu, China.
Mol Ther Methods Clin Dev. 2024 Dec 17;33(1):101399. doi: 10.1016/j.omtm.2024.101399. eCollection 2025 Mar 13.
The multi-administration of recombinant adeno-associated virus (rAAV) is limited largely by immunological barriers. Herein, a novel strategy, named rAAV pseudo-lipid nanoparticle combined with triamcinolone acetonide (LNP-rAAV + TAC), has been described in mice. We showed successful but low efficient triple trafficking by LNP-rAAV2 carrying EGFP, human factor IX (hFIX), and luciferase (luc), due to its encapsulation characteristic. Additionally, sustained TAC treatment, which dose-dependently downregulated the anti-rAAV2 antibodies, permitted rAAV2 re-administration at dosages of ≥45 mg/kg/3 days. Furthermore, to improve the efficiency and safety, LNP-rAAV + TAC was evaluated, using LNP-rAAV2 carrying EGFP, hFIX, and luc co-treating with 45 mg/kg/3 days TAC before and after treatment with LNP-rAAV2 injections. Notable neutralizing antibody reductions of 37.8-fold and 12.7-fold were observed by the combinatorial strategy compared with the independent LNP encapsulation and TAC treatment approaches. The plasma hFIX protein was enhanced to 15.1 μg/mL and the liver bioluminescence was elevated to 1.4 × 10 p/s/cm/sr following the second and third administrations, with weaker levels in LNP encapsulation (1.9 μg/mL, 2.1 × 10 p/s/cm/sr) and TAC treatment (3.0 μg/mL, 6.1 × 10 p/s/cm/sr) groups. Thus, this combination strategy is an attractive candidate for enabling multi-dosing of rAAV vector and warrants further study on the underlying mechanism.
重组腺相关病毒（rAAV）的多次给药在很大程度上受到免疫屏障的限制。在此，在小鼠中描述了一种名为rAAV假脂质纳米颗粒联合曲安奈德（LNP-rAAV + TAC）的新策略。我们发现携带增强型绿色荧光蛋白（EGFP）、人凝血因子IX（hFIX）和荧光素酶（luc）的LNP-rAAV2实现了成功但低效的三重转运，这归因于其封装特性。此外，持续的曲安奈德治疗可剂量依赖性地下调抗rAAV2抗体，从而允许以≥45 mg/kg/3天的剂量重新给予rAAV2。此外，为了提高效率和安全性，对LNP-rAAV + TAC进行了评估，方法是在LNP-rAAV2注射治疗前后，使用携带EGFP、hFIX和luc的LNP-rAAV2与45 mg/kg/3天的曲安奈德联合治疗。与独立的LNP封装和曲安奈德治疗方法相比，联合策略使中和抗体显著降低了37.8倍和12.7倍。在第二次和第三次给药后，血浆hFIX蛋白增加到15.1 μg/mL，肝脏生物发光增加到1.4×10 p/s/cm/sr，而在LNP封装组（1.9 μg/mL，2.1×10 p/s/cm/sr）和曲安奈德治疗组（3.0 μg/mL，6.1×10 p/s/cm/sr）中水平较低。因此，这种联合策略是实现rAAV载体多次给药的有吸引力的候选方案，值得进一步研究其潜在机制。
