rAAV6-mediated miR-29b delivery suppresses renal fibrosis
rAAV6-mediated miR-29b delivery suppresses renal fibrosis
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DOI：
10.1007/s10157-019-01783-w
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发表时间：
2019-09
影响因子：
2.3
通讯作者：
Suguru Saito;Shin-ichiro Ohno;Yuichirou Harada;K. Oikawa;Koji Fujita;Shouichirou Mineo;Asako Gondo;Y. Kanno;M. Kuroda
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文献类型：
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作者：
Suguru Saito;Shin-ichiro Ohno;Yuichirou Harada;K. Oikawa;Koji Fujita;Shouichirou Mineo;Asako Gondo;Y. Kanno;M. Kuroda
背景研究表明microRNA-29 b（miR-29 b）抑制肾纤维化。因此，miR-29 b替代疗法代表了治疗肾纤维化的有希望的方法。然而，尚未建立肾靶向miRNA递送的有效方法。重组腺相关病毒（rAAV）载体具有巨大的临床应用潜力。对于肾靶向基因递送，尚未建立最合适的AAV血清型。在这里，我们确定了最合适的AAV血清型的肾靶向基因传递，并确定，AAV介导的miR-29 b传递可以抑制肾纤维化invivo.MethodTo确定哪种AAV血清型是适合肾细胞，GFP阳性细胞通过流式细胞术后感染的rAAV血清型1-9载体含有theEGFP基因。接下来，我们将rAAV载体注射到肾盂中以确定体内转导效率。在注射携带GFP基因的rAAV血清型1-9载体后7天测量GFP表达。最后，我们研究了rAAV 6介导的miR-29 b递送是否可以抑制UUO小鼠模型中的肾纤维化。ResultsWe发现，rAAV 6载体是最适合靶向肾细胞，无论动物物种在体外和rAAV 6是最适合的载体，在小鼠体内肾靶向基因递送。肾盂内注射rAAV载体可将基因导入肾TEC。结论rAAV 6是最适合于肾脏靶向基因转染的血清型，rAAV 6介导的miR-29 b转染入肾TEC可抑制已建立的肾纤维化。
BackgroundPrevious 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.MethodTo 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 theEGFPgene. 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 theEGFPgene. Finally, we investigated whether rAAV6-mediated miR-29b delivery can suppress renal fibrosis in UUO mouse model.ResultsWe 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 suppressingSnail1expression.ConclusionOur 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.