Casein kinase 2 activity is a host restriction factor for AAV transduction

Area of Science:

• Molecular Biology
• Gene Therapy
• Cardiovascular Research

Background:

• Adeno-associated virus (AAV) vectors are crucial for gene therapy, but their transduction efficiency, especially in the human heart, is limited by poorly understood cellular mechanisms.
• Developing effective gene therapies requires overcoming these barriers to optimize vector delivery and expression.

Purpose of the Study:

• To identify and overcome cellular barriers limiting AAV transduction in the heart.
• To investigate the role of casein kinase 2 (CK2) signaling in AAV vector-host interactions.

Main Methods:

• Utilized induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iPSC-CMs), cardiac fibroblasts (iPSC-CFs), and primary endothelial cells to model AAV transduction.
• Performed phosphoproteome analysis to identify affected cellular pathways.
• Assessed the impact of transient CK2 inhibition on AAV transduction rates and transgene expression.

Main Results:

• Phosphoproteome analysis revealed CK2 signaling as a significantly affected pathway upon AAV exposure.
• Transient inhibition of CK2 activity substantially enhanced transduction rates for AAV2, AAV6, and AAV9 across all tested cardiac cell types.
• CK2 inhibition improved cytoplasmic AAV trafficking, modulated DNA damage response by destabilizing MRE11, and altered RNA processing pathways.
• CK2 inhibition augmented transgene expression in iPSC-CFs, suggesting a role in regulating latent AAV genomes.

Conclusions:

• CK2 activity represents a key cellular barrier to efficient AAV transduction and transgene expression in cardiac cells.
• Targeting CK2 offers a promising strategy to improve the efficacy of AAV-based gene therapies for cardiovascular applications.