Efficient kidney gene transfer and proximal tubule transduction using self-complementary AAV.cc47 vectors

Area of Science:

• * Molecular and Cellular Biology
• * Gene Therapy
• * Virology

Background:

• * Gene delivery to kidney cells is crucial for developing gene therapies for kidney diseases.
• * Adeno-associated virus (AAV) vectors are promising tools for gene therapy.
• * The AAV.cc47 variant, evolved for improved transduction, has not been fully explored in the kidney.

Purpose of the Study:

• * To evaluate the kidney transduction efficiency of the novel AAV.cc47 vector.
• * To compare AAV.cc47 with AAV9 in kidney cells, including human kidney organoids.
• * To develop a method for quantifying transgene expression in specific kidney cell types.

Main Methods:

• * In vivo studies in mice and in vitro studies using human kidney organoids.
• * Systemic administration of AAV.cc47 and AAV9 vectors.
• * Quantitative analysis of transgene expression using automated cell classification.

Main Results:

• * AAV.cc47 demonstrated robust transduction in mouse kidneys and human kidney organoids, primarily in proximal tubule (PT) epithelium.
• * AAV.cc47 showed higher kidney transduction efficiency (>80% PT epithelium) compared to AAV9 at low vector doses.
• * Similar biodistribution was observed for AAV.cc47 and AAV9 in renal and extrarenal tissues.
• * Self-complementary AAV.cc47 vectors transduced a subset of PT epithelium with no detectable transduction in non-PT cells.
• * Dual AAV.cc47 vectors were shown to increase genome payload capacity for kidney gene transfer.

Conclusions:

• * AAV.cc47 is a highly efficient vector for gene delivery to kidney proximal tubule epithelium.
• * The developed quantitative analysis method can be adapted for evaluating other AAV vectors and kidney cell types.
• * AAV.cc47 holds significant potential as a research tool and for clinical applications in kidney disease gene therapy.