SNAC: A Single-Nuclei Atlas of Capsid Distribution in Nonhuman Primate Eye

Area of Science:

• Gene Therapy
• Molecular Biology
• Bioinformatics

Background:

• Adeno-associated virus (AAV) vectors are leading gene therapy tools, with approved therapies like Luxturna and Zolgensma.
• Natural and engineered AAV capsids require comprehensive biodistribution profiling for improved cell-type specificity.
• Current single-cell methods face challenges in detecting low AAV transduction levels, hindering capsid screening.

Purpose of the Study:

• To develop an improved method for single-nuclei profiling of AAV transduction at multiplex scale.
• To enable accurate cell-type-specific biodistribution analysis of AAV capsids.
• To reduce the time, effort, and cost associated with AAV capsid profiling.

Main Methods:

• Development of SNAC (Single-Nuclei Atlas of Capsid distribution), a novel single-nuclei profiling technique.
• Application of SNAC to a nonhuman primate eye model for proof of concept.
• Quantification of vector expression across major retinal cell types using SNAC.

Main Results:

• SNAC accurately identifies and quantifies AAV vector expression in all major retinal cell types.
• Capsid rankings obtained via SNAC align with results from established tissue sampling protocols.
• The method demonstrates feasibility for multiplex-scale single-nuclei profiling of AAV transduction.

Conclusions:

• SNAC is an effective method for high-resolution AAV biodistribution profiling.
• This technique enhances the ability to refine AAV cell-type specificity and minimize off-target effects in gene therapy.
• SNAC offers a promising solution for efficient and cost-effective AAV capsid screening.