RNAi-nanoparticulate manipulation of gene expression as a new functional genomics tool in the liver

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• 作者：Hao Yin¹ ; ^&dagger ; ; Roman L. Bogorad¹ ; ^&dagger ; ; Carmen Barnes² ; Stephen Walsh¹ ; Iris Zhuang¹ ; ³ ; Hidenori Nonaka⁴ ; Vera Ruda¹ ; Satya Kuchimanchi² ; Lubomir Nechev² ; Akin Akinc² ; Wen Xue⁵ ; Marino Zerial⁴ ; Robert Langer¹ ; ⁶ ; ⁷ ; ⁸ ; Daniel G. Anderson¹ ; ⁶ ; ⁷ ; ⁸ ; ^&Dagger ; ; ^{dgander@mit.edu" class="auth_mail" title="E-mail the corresponding author} ; Victor Koteliansky⁹ ; ¹⁰ ; ^&Dagger ; ; ^{V.Kotelianski@Skoltech.RU" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Hippo pathway ; p53 ; Bile acids ; siRNA ; Nanoparticles
• 刊名：Journal of Hepatology
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：64
• 期：4
• 页码：899-907
• 全文大小：2524 K

文摘

The Hippo pathway controls organ size through a negative regulation of the transcription co-activator Yap1. The overexpression of hyperactive mutant Yap1 or deletion of key components in the Hippo pathway leads to increased organ size in different species. Analysis of interactions of this pathway with other cellular signals corroborating organ size control is limited in part due to the difficulties associated with development of rodent models.

Methods

Here, we develop a new model of reversible induction of the liver size in mice using siRNA-nanoparticles targeting two kinases of the Hippo pathway, namely, mammalian Ste20 family kinases 1 and 2 (Mst1 and Mst2), and an upstream regulator, neurofibromatosis type II (Nf2).

Results

The triple siRNAs nanoparticle-induced hepatomegaly in mice phenocopies one observed with Mst1^−/−Mst2^−/− liver-specific depletion, as shown by extensive proliferation of hepatocytes and activation of Yap1. The simultaneous co-treatment with a fourth siRNA nanoparticle against Yap1 fully blocked the liver growth. Hippo pathway-induced liver enlargement is associated with p53 activation, evidenced by its accumulation in the nuclei and upregulation of its target genes. Moreover, injections of the triple siRNAs nanoparticle in p53^LSL/LSL mice shows that livers lacking p53 expression grow faster and exceed the size of livers in p53 wild-type animals, indicating a role of p53 in controlling Yap1-induced liver growth.

Conclusion

Our data show that siRNA-nanoparticulate manipulation of gene expression can provide the reversible control of organ size in adult animals, which presents a new avenue for the investigation of complex regulatory networks in liver.