Li Mei, Ma Haixiao, Wu Yang, Gao Yunling, Wang Jie, Wang Hanbing
Department of Anesthesiology, First People Hospital of Foshan, Foshan, China.
Institute of Neuroscience and Brain Diseases; Xiangyang Central Hospital, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang, China.
Front Med (Lausanne). 2025 Apr 4;12:1554449. doi: 10.3389/fmed.2025.1554449. eCollection 2025.
Goat-derived adeno-associated virus (AAV) vectors, such as AAV Go.1, represent a novel platform for gene therapy due to their unique origin and potential advantages in transduction efficiency and immune evasion. However, their therapeutic potential and biological properties remain underexplored.
In this study, we developed a recombinant AAV (rAAV) Go.1 by replacing the goat AAV rep gene with the standard AAV2-rep gene to improve packaging efficiency. We compared the transduction efficiency of rAAV Go.1 with that of AAV5, a closely related serotype with 95% genome similarity, both and . Additionally, we assessed immune evasion properties by evaluating resistance to neutralization using sera from rAAV5-immunized mice and human volunteers. To further enhance transduction efficiency, we introduced site-specific mutations in the VP1 unique (VP1u) region and VP1/2 common region.
The rep gene modification led to a significantly higher packaging efficiency for rAAV Go.1 compared to the original goat AAV. rAAV Go.1 exhibited markedly higher transduction efficiency than AAV5 in both in vitro and in vivo models. Furthermore, rAAV Go.1 demonstrated a 4-fold increase in resistance to neutralization by sera from rAAV5-immunized mice. A study involving 20 healthy volunteers revealed that high-titer neutralizing antibodies had a more pronounced inhibitory effect on rAAV5 compared to rAAV Go.1. Mutagenesis studies identified key modifications that enhanced viral properties: K32R, K91R, and K122R mutations in the VP1u region significantly improved viral production, while K137R (VP1u) enhanced transduction efficiency and .
Our findings highlight the potential of rAAV Go.1 as an improved gene therapy vector with superior transduction efficiency and enhanced immune evasion. The identified VP1 mutations further optimize viral properties, making rAAV Go.1 a promising candidate for future therapeutic applications.
源自山羊的腺相关病毒（AAV）载体，如AAV Go.1，由于其独特的起源以及在转导效率和免疫逃逸方面的潜在优势，代表了一种新型的基因治疗平台。然而，它们的治疗潜力和生物学特性仍未得到充分探索。
在本研究中，我们通过用标准的AAV2-rep基因替换山羊AAV rep基因来提高包装效率，从而构建了重组AAV（rAAV）Go.1。我们比较了rAAV Go.1与AAV5（一种与之密切相关、基因组相似度达95%的血清型）在体外和体内模型中的转导效率。此外，我们通过评估rAAV5免疫小鼠和人类志愿者血清对中和作用的抗性来评估免疫逃逸特性。为了进一步提高转导效率，我们在VP1独特（VP1u）区域和VP1/2共同区域引入了位点特异性突变。
与原始山羊AAV相比，rep基因修饰使rAAV Go.1的包装效率显著提高。在体外和体内模型中，rAAV Go.1均表现出比AAV5明显更高的转导效率。此外，rAAV Go.1对rAAV5免疫小鼠血清中和作用的抗性提高了4倍。一项涉及20名健康志愿者的研究表明，高滴度中和抗体对rAAV5的抑制作用比对rAAV Go.1更为明显。诱变研究确定了增强病毒特性的关键修饰：VP1u区域的K32R、K91R和K122R突变显著提高了病毒产量，而K137R（VP1u）增强了转导效率。
我们的研究结果突出了rAAV Go.1作为一种改进的基因治疗载体的潜力，其具有卓越的转导效率和增强的免疫逃逸能力。所确定的VP1突变进一步优化了病毒特性，使rAAV Go.1成为未来治疗应用的有希望的候选者。
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