RNA-Seq Analysis of Trans-Differentiated ARPE-19 Cells Transduced by AAV9-AIPL1 Vectors

Area of Science:

• Ophthalmology
• Gene Therapy
• Molecular Biology

Background:

• Inherited retinal disorders (IRDs) are a significant cause of blindness, with gene therapy emerging as a promising treatment modality.
• Adeno-associated virus (AAV)-based gene therapy has shown success, notably for Leber congenital amaurosis type 2 (LCA2).
• Aryl hydrocarbon receptor-interacting protein like-1 (AIPL1) mutations cause LCA4, a form of IRD.

Purpose of the Study:

• To design, produce, and in vitro test an AAV serotype 9 (AAV9) vector carrying a codon-optimized AIPL1 gene (AAV9-AIPL1co) for potential LCA4 treatment.
• To evaluate the immunogenicity and gene expression profile of AAV9-AIPL1co compared to wild-type AIPL1 (AAV9-AIPL1wt).

Main Methods:

• Development and production of the pAAV-AIPL1co vector.
• Transduction of retinal pigment epithelium (ARPE-19) cells with AAV9-AIPL1co and AAV9-AIPL1wt.
• In vitro assessment of AIPL1 expression and cellular antiviral response using RNA-sequencing (RNA-seq).

Main Results:

• AAV9-AIPL1co successfully transduced ARPE-19 cells and initiated human AIPL1 expression.
• Cells transduced with AAV9-AIPL1co exhibited a significantly reduced antiviral response compared to those with AAV9-AIPL1wt.
• RNA-seq analysis revealed distinct gene expression profiles, with AAV9-AIPL1co showing less activation of interferon-stimulated genes and innate immune response pathways.

Conclusions:

• The codon-optimized AAV9-AIPL1co vector demonstrates reduced immunogenicity in vitro.
• This suggests AAV9-AIPL1co is a potential candidate for developing gene therapies for LCA4.
• Further studies using animal models are warranted to assess the therapeutic efficacy and safety of AAV9-AIPL1co.