基于重组腺相关病毒（rAAV）的病毒载体已成为中枢神经系统（CNS）中使用最广泛的治疗性基因递送系统。尽管在神经学应用中具有临床安全性和有效性，但采用当前一代载体的障碍在于其效率有限，导致中枢神经系统靶细胞的转导有限。为了解决这一限制，研究人员通过生物工程设计了适合用途的 AAV，具有改进的中枢神经系统趋向性和组织穿透力。虽然这些新型 AAV 的临床前评估主要在动物模型中进行，但人类诱导多能干细胞 (hiPSC) 衍生的类器官为在人类环境中对新型 AAV 变体进行功能评估提供了独特的机会。在这项研究中，我们对大量野生型和生物工程 AAV 衣壳在 hiPSC 衍生的脑类器官中的转导效率进行了全面、公正的评估。我们证明，在类器官中观察到的有效 AAV 转导在脑脊液 (CSF) 输送后在小鼠和非人灵长类动物模型中得到了体内重现。总之，我们的研究展示了使用大脑类器官系统来预筛选新型 AAV 载体。此外，我们还报告了新型 AAV 变体的数据，这些变体在体内临床前模型中通过 CSF 传递时表现出更高的 CNS 转导效率。© 2024 作者。

Viral vectors based on recombinant adeno-associated virus (rAAV) have become the most widely used system for therapeutic gene delivery in the central nervous system (CNS). Despite clinical safety and efficacy in neurological applications, a barrier to adoption of the current generation of vectors lies in their limited efficiency, resulting in limited transduction of CNS target cells. To address this limitation, researchers have bioengineered fit-for-purpose AAVs with improved CNS tropism and tissue penetration. While the preclinical assessment of these novel AAVs is primarily conducted in animal models, human induced pluripotent stem cell (hiPSC)-derived organoids offer a unique opportunity to functionally evaluate novel AAV variants in a human context. In this study, we performed a comprehensive and unbiased evaluation of a large number of wild-type and bioengineered AAV capsids for their transduction efficiency in hiPSC-derived brain organoids. We demonstrate that efficient AAV transduction observed in organoids was recapitulated in vivo in both mouse and non-human primate models after cerebrospinal fluid (CSF) delivery. In summary, our study showcases the use of brain organoid systems for the pre-screening of novel AAV vectors. Additionally, we report data for novel AAV variants that exhibit improved CNS transduction efficiency when delivered via the CSF in in vivo preclinical models.© 2024 The Authors.