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In vivo delivery of transcription factors with multifunctional oligonucleotides

Nature Materials volume 14, pages 701–706 (2015)Cite this article

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Abstract

Therapeutics based on transcription factors have the potential to revolutionize medicine but have had limited clinical success as a consequence of delivery problems1,2,3,4. The delivery of transcription factors is challenging because it requires the development of a delivery vehicle that can complex transcription factors, target cells and stimulate endosomal disruption, with minimal toxicity5,6. Here, we present a multifunctional oligonucleotide, termed DARTs (DNA assembled recombinant transcription factors), which can deliver transcription factors with high efficiency in vivo. DARTs are composed of an oligonucleotide that contains a transcription-factor-binding sequence and hydrophobic membrane-disruptive chains that are masked by acid-cleavable galactose residues. DARTs have a unique molecular architecture, which allows them to bind transcription factors, trigger endocytosis in hepatocytes, and stimulate endosomal disruption. The DARTs have enhanced uptake in hepatocytes as a result of their galactose residues and can disrupt endosomes efficiently with minimal toxicity, because unmasking of their hydrophobic domains selectively occurs in the acidic environment of the endosome. We show that DARTs can deliver the transcription factor nuclear erythroid 2-related factor 2 (Nrf2) to the liver, catalyse the transcription of Nrf2 downstream genes, and rescue mice from acetaminophen-induced liver injury.

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Figure 1: DARTs are multifunctional oligonucleotides that can deliver the transcription factor Nrf2 in vivo.
Figure 2: The DARTs are acid-sensitive endosomal-disruptive agents and their hydrolysis is pH sensitive.
Figure 3: The DARTs can target hepatocytes and enhance the delivery of Nrf2 to the liver.
Figure 4: DARTs are able to deliver Nrf2 to hepatocytes, upregulate Nrf2 downstream genes, and can protect hepatocytes against reactive oxygen species (ROS).
Figure 5: The DARTs can deliver Nrf2 to the liver, upregulate Nrf2 downstream genes, and rescue mice from APAP-induced liver injury.

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Acknowledgements

This work was supported by grants from the NIH, U01 268201000043C-0-0-1, RO1 AI107116-01 and RO1 AI088023-03. In addition, this work was supported by the WM Keck Foundation, based in Los Angeles, USA. We thank S. Maity and M. Webb for chemistry advice, and M. West in the Berkeley Stem Cell Center for technical assistance.

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Author notes

  1. Kunwoo Lee, Mohammad Rafi and Xiaojian Wang: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA,

    Kunwoo Lee, Mohammad Rafi, Xiaojian Wang, Kiana Aran, Xuli Feng, Carlo Lo Sterzo, Richard Tang, Nithya Lingampalli, Hyun Jin Kim & Niren Murthy

  2. UC Berkeley and UCSF Joint Graduate Program in Bioengineering, Berkeley/San Francisco, USA

    Kunwoo Lee & Niren Murthy

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  1. Kunwoo Lee
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Contributions

K.L., M.R., X.W., X.F. and N.M. designed the experiments. X.W. and X.F. carried out organic synthesis. K.L., M.R., X.F., C.S, R.T. and N.L. performed characterization, in vitro, and in vivo experiments. All of the authors were involved in the analyses and interpretation of data. K.L., M.R., X.W. and N.M. wrote the paper, with the help of the co-authors.

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Correspondence to Niren Murthy.

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The authors declare no competing financial interests.

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Lee, K., Rafi, M., Wang, X. et al. In vivo delivery of transcription factors with multifunctional oligonucleotides. Nature Mater 14, 701–706 (2015). https://doi.org/10.1038/nmat4269

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