Optimal different adeno-associated virus capsid/promoter combinations to target specific cell types in the common marmoset cerebral cortex

Yasunori Matsuzaki; Yuuki Fukai; Ayumu Konno; Hirokazu Hirai

doi:10.1016/j.omtm.2024.101337

Optimal different adeno-associated virus capsid/promoter combinations to target specific cell types in the common marmoset cerebral cortex

Mol Ther Methods Clin Dev. 2024 Sep 13;32(4):101337. doi: 10.1016/j.omtm.2024.101337. eCollection 2024 Dec 12.

Authors

Yasunori Matsuzaki^{1

2}, Yuuki Fukai¹, Ayumu Konno^{1

2}, Hirokazu Hirai^{1

2}

Affiliations

¹ Department of Neurophysiology & Neural Repair, Gunma University Graduate School of Medicine, Maebashi, Gunma 371-8511, Japan.
² Viral Vector Core, Gunma University, Initiative for Advanced Research, Maebashi, Gunma 371-8511, Japan.

Abstract

To achieve cell-type-specific gene expression, using target cell-type-tropic different adeno-associated virus (AAV) capsids is advantageous. However, their tropism across brain cell types in nonhuman primates has not been fully elucidated. We assessed the tropism of nine AAV serotype capsids (AAV1, 2, 5, 6, 7, 8, 9, rh10, and DJ) expressing EGFP by chicken β-actin hybrid (CBh) promoter in marmoset cerebral cortical cells. All nine AAV capsid vectors, especially AAV9 and AAVrh10, caused highly neuron-selective EGFP expression. Some AAV capsids, including AAV5, induced EGFP expression to a lesser extent in oligodendrocytes. Different ubiquitous cytomegalovirus (CMV) and CMV early enhancer/chicken β-actin (CAG) promoters exhibited similar neuron-predominant transgene expression. Conversely, all nine AAV capsid vectors with the astrocyte-specific hGFA(ABC1D) promoter selectively expressed EGFP in astrocytes, except AAV5, which modestly expressed EGFP in oligodendrocytes. Oligodendrocyte-specific mouse myelin basic protein (mMBP) promoter in AAV5 vectors expressed EGFP in oligodendrocytes specifically and efficiently. The following are optimal combinations of capsids and promoters for cell-type-specific expression: AAV9 or AAVrh10 and ubiquitous CBh or CMV promoter for neuron-specific transgene expression, AAV2 or AAV7 and hGFA(ABC1D) promoters for astrocyte-specific transgene expression, and AAV5 and mMBP promoters for oligodendrocyte-specific transgene expression.

Keywords: adeno-associated virus; astrocytes; capsid; cell-type-specific promoter; glial fibrillary acidic protein; marmosets; microglia; myelin basic protein; oligodendrocytes; serotype.