Assessment of Adipocyte Transduction Using Different AAV Capsid Variants

Area of Science:

• Gene therapy
• Viral vector technology
• Cell biology

Background:

• Adeno-associated viruses (AAVs) are critical tools for gene delivery in mammalian cells.
• Evaluating AAV serotype efficacy is essential for optimizing gene therapy strategies.

Purpose of the Study:

• To compare the transduction efficiency of AAV2/5, AAV2/6, AAV2/8, and AAV2/9 vectors expressing GFP in preadipocyte and mature adipocyte cells.
• To assess the in vivo transduction capacity of selected AAV serotypes in mouse adipose tissue.

Main Methods:

• Utilized live imaging microscopy (IncuCyte S3) and flow cytometry to quantify GFP expression.
• Assessed transduction in 3T3-L1 preadipocytes, differentiating cells, and mature adipocytes.
• Conducted in vivo studies by injecting AAV vectors into the adipose tissue of C57BL6 mice.

Main Results:

• AAV2/6 exhibited 1.5-2 fold higher transduction efficiency in 3T3-L1 preadipocytes than AAV2/5 and AAV2/8.
• GFP expression remained stable for up to 20 days, unaffected by adipocyte differentiation.
• AAV2/6 demonstrated effective transduction of mature adipocytes and showed the highest in vivo transducing activity in mouse adipose tissue.

Conclusions:

• AAV2/6 is a highly effective vector for gene delivery to both preadipocytes and mature adipocytes.
• AAV2/6, alongside AAV2/8 and AAV2/9, offers promising options for adipocyte-targeted gene therapy.