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rAAV6-mediated miR-29b delivery suppresses renal fibrosis

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Abstract

Background

Previous studies showed that microRNA-29b (miR-29b) inhibits renal fibrosis. 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. 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.

Method

To determine which AAV serotype is suitable for kidney cells, GFP-positive cells were identified by flow cytometry after the infection of rAAV serotype 1–9 vectors containing the EGFP gene. Next, we injected rAAV vectors into the renal pelvis to determine transduction efficiency in vivo. GFP expression was measured seven days after injecting rAAV serotype 1–9 vectors carrying the EGFP gene. Finally, we investigated whether rAAV6-mediated miR-29b delivery can suppress renal fibrosis in 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

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.

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Acknowledgements

We thank Dr. Riichi Takahashi and Dr. Motohito Goto of Central Institute for Experimental Animals (CIEA) for supporting our in vivo analysis. This research was supported by a Grants-in-Aid for scientific research (B: 17H04067) and (17K16101) and by the Development program of microRNA measurement technology foundation in body fluid from Japan Agency for Medical Research and development, AMED and supported in part by Strategic Research Foundation Grant-aided Project for Private Universities from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Authors and Affiliations

  1. Department of Nephrology, Tokyo Medical University, Tokyo, Japan

    Suguru Saito, Asako Gondo & Yoshihiko Kanno

  2. Deparatment of Molecular Pathology, Tokyo Medical University, 6-1-1 Shinjyuku, Shinjyuku-ku, Tokyo, 160-8402, Japan

    Shin-ichiro Ohno, Yuichirou Harada, Keiki Oikawa, Koji Fujita, Shouichirou Mineo & Masahiko Kuroda

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  1. Suguru Saito
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Correspondence to Shin-ichiro Ohno.

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Saito, S., Ohno, Si., Harada, Y. et al. rAAV6-mediated miR-29b delivery suppresses renal fibrosis. Clin Exp Nephrol 23, 1345–1356 (2019). https://doi.org/10.1007/s10157-019-01783-w

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  • DOI: https://doi.org/10.1007/s10157-019-01783-w

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