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Abstract: TH-PO961

rAAV6-Mediated miR-29b Delivery Suppresses Renal Fibrosis

Session Information

Category: Pathology and Lab Medicine

  • 1501 Pathology and Lab Medicine: Basic

Authors

  • Saito, Suguru, Tokyo Medical University, Tokyo, Japan
  • Ohno, Shinichiro, Tokyo Medical University, Tokyo, Japan
  • Kuroda, Masahiko, Tokyo Medical University, Tokyo, Japan
  • Kanno, Yoshihiko, Tokyo Medical University, Tokyo, Japan
Background

Renal fibrosis is a common feature of CKD, which has no available specific treatment. Micro RNAs (miRNAs) are short endogenous non-coding RNAs that regulate various cellular processes. Previous studies showed that miR-29b inhibits renal fibrosis in mice models. Therefore, miR-29b replacement therapy represents a promising approach for treating renal fibrosis. However, an efficient method of kidney-targeted miRNA delivery has yet to be established.
Recombinant adeno-associated virus (rAAV) vectors have great potential for clinical application. The distinct AAV serotypes exhibit different tissue tropisms. For kidney-targeted gene delivery, the most suitable AAV serotype has yet to be established. Here, we identified the most suitable AAV serotype for kidney-targeted gene delivery, and determined that AAV-mediated miR-29b delivery can suppress renal fibrosis in vivo.

Methods

We used the normal rat kidney fibroblast cell line (NRK-49F), rat TEC line (NRK-52E), and human kidney proximal TECs to determine which AAV serotype is suitable for kidney-targeted gene delivery in vitro. To determine transduction efficiency, GFP-positive cells were identified by flow cytometry after the infection of rAAV serotype 1–9 vectors containing the EGFP gene.
Next, we went on to investigate the transduction efficiency of mouse kidney by using rAAV serotype 1–9 vectors in vivo. We injected rAAV vectors into the renal pelvis. To determine transduction efficiency, GFP expression was measured seven days after injecting rAAV vectors carrying the EGFP gene
Finally, we investigated whether rAAV6-mediated miR-29b delivery can suppress renal fibrosis in a unilateral ureteral obstruction (UUO) mouse model.

Results

We found that rAAV6 vector is the most suitable for targeting kidney cells regardless of animal species in vitro and rAAV6 is the most suitable vector for kidney-targeted in vivo gene delivery in mice. Intra-renal pelvic injection of rAAV vectors can transduce genes into kidney TECs. Furthermore, rAAV6-mediated miR-29b delivery attenuated renal fibrosis in UUO model by suppressing Snail1 expression.

Conclusion

In summary, our study has revealed that rAAV6 is the most suitable serotype for kidney-targeted gene delivery and rAAV6-mediated miR-29b delivery into kidney TECs can suppress established renal fibrosis. Our findings may provide novel information for facilitating kidney gene therapy.

Funding

  • Government Support - Non-U.S.