Surdyka Magdalena, Jesion Ewelina, Niewiadomska-Cimicka Anna, Trottier Yvon, Kalinowska-Pośka Żaneta, Figiel Maciej
Department of Molecular Neurobiology, Institute of Bioorganic Chemistry Polish Academy of Sciences, Poznań, Poland.
Institute of Genetics and Molecular and Cellular Biology, INSERM U1258, CNRS UMR7104, University of Strasbourg, Illkirch, France.
Front Mol Neurosci. 2022 Sep 13;15:947490. doi: 10.3389/fnmol.2022.947490. eCollection 2022.
Adeno-associated virus (AAV)-based brain gene therapies require precision without off-targeting of unaffected neurons to avoid side effects. The cerebellum and its cell populations, including granule and Purkinje cells, are vulnerable to neurodegeneration; hence, conditions to deliver the therapy to specific cell populations selectively remain challenging. We have investigated a system consisting of the AAV serotypes, targeted injections, and transduction modes (direct or retrograde) for targeted delivery of AAV to cerebellar cell populations. We selected the AAV-PHP.eB and AAVrh10 serotypes valued for their retrograde features, and we thoroughly examined their cerebellar transduction pattern when injected into lobules and deep cerebellar nuclei. We found that AAVrh10 is suitable for the transduction of neurons in the mode highly dependent on placing the virus at axonal terminals. The strategy secures selective transduction for granule cells. The AAV-PHP.eB can transduce Purkinje cells and is very selective for the cell type when injected into the DCN at axonal PC terminals. Therefore, both serotypes can be used in a retrograde mode for selective transduction of major neuronal types in the cerebellum. Moreover, our transduction strategies are suitable for pre-clinical protocol development for gene delivery to granule cells by AAVrh10 and Purkinje cells by AAV-PHPeB.
基于腺相关病毒(AAV)的脑基因治疗需要精准性,避免对未受影响的神经元产生脱靶效应以避免副作用。小脑及其细胞群体,包括颗粒细胞和浦肯野细胞,易发生神经退行性变;因此,将治疗选择性地递送至特定细胞群体的条件仍然具有挑战性。我们研究了一个由AAV血清型、靶向注射和转导模式(顺行或逆行)组成的系统,用于将AAV靶向递送至小脑细胞群体。我们选择了因其逆行特性而有价值的AAV-PHP.eB和AAVrh10血清型,并在将它们注射到小叶和小脑深部核团时,全面检查了它们的小脑转导模式。我们发现AAVrh10适合以高度依赖于将病毒置于轴突末端的模式转导神经元。该策略确保了对颗粒细胞的选择性转导。当在轴突浦肯野细胞末端注射到深部小脑核时,AAV-PHP.eB可以转导浦肯野细胞,并且对细胞类型具有高度选择性。因此,两种血清型都可以以逆行模式用于小脑主要神经元类型的选择性转导。此外,我们的转导策略适用于临床前方案的开发,用于通过AAVrh10将基因递送至颗粒细胞以及通过AAV-PHPeB将基因递送至浦肯野细胞。