Two engineered AAV capsid variants for efficient transduction of human cortical neurons directly converted from iPSC

• Published in: Journal of neuroscience methods Vol. 368; p. 109457
• Main Authors: Fischer, Sandra, Strobel, Benjamin, Weinmann, Jonas, Gillardon, Frank
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.02.2022
• Subjects: Adeno-associated virus; Animals; Capsid - metabolism; Capsid variants; Dependovirus - genetics; Genetic Vectors; Humans; Induced human neurons; Induced Pluripotent Stem Cells; Neuronal function; Neurons; Rats; Transduction efficiency; Transduction, Genetic; Induced human neurons; Adeno-associated virus; Capsid variants; Transduction efficiency; Neuronal function
• ISSN: 0165-0270; 1872-678X
• DOI: 10.1016/j.jneumeth.2021.109457

Recombinant adeno-associated virus (AAV) is the most widely used vector for gene therapy in clinical trials. To increase transduction efficiency and specificity, novel engineered AAV variants with modified capsid sequences are evaluated in human cell cultures and non-human primates. We tested two novel AAV capsid variants, AAV2-NNPTPSR and AAV9-NVVRSSS, in human cortical neurons, which were directly converted from human induced pluripotent stem cells and cocultured with rat primary astrocytes. AAV2-NNPTPSR variant efficiently transduced both induced human cortical glutamatergic neurons and induced human cortical GABAergic interneurons. By contrast, AAV9-NVVRSSS variant transduced both induced human cortical neurons and cocultured rat primary astrocytes. High viral titers (1E+5 viral genomes per cell) caused a significant decrease in viability of induced human cortical neurons. Low viral titers (1E+4 viral genomes per cell) led to a significant increase in the neuronal activity marker c-Fos in transduced human neurons following treatment with a potassium channel blocker. We identified two engineered AAV capsid variants that efficiently transduce induced human cortical neurons. The threefold higher percentage of c-Fos positive, transduced human neurons may indicate functional alterations induced by viral transduction and/or transgene expression. •AAV2-NNPTPSR capsid variant efficiently transduces induced human cortical neurons.•AAV9-NVVRSSS capsid variant transduces both induced human neurons and rat astrocytes.•1E+5 viral genomes per cell cause a significant decrease in neuronal viability.•1E+4 viral genomes per cell seem to affect neuronal function.