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Albumin-Binding Aptamer Chimeras for Improved siRNA Bioavailability

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Abstract

Introduction

Short interfering RNAs (siRNAs) are potent nucleic acid-based drugs designed to target disease driving genes that may otherwise be undruggable with small molecules. However, therapeutic potential of siRNA in vivo is limited by poor pharmacokinetic properties, including rapid renal clearance and nuclease degradation. Backpacking on natural carriers such as albumin, which is present at high concentration and has a long half-life in serum, is an effective way to modify pharmacokinetics of biologic drugs that otherwise have poor bioavailability. In this work, we sought to develop albumin-binding aptamer-siRNA chimeras to improve the bioavailability of siRNA.

Methods

A Systematic Evolution of Ligands through Exponential Enrichment (SELEX) approach was used to obtain modified RNA-binding aptamers, which were then fused directly to siRNA via in vitro transcription. Molecular and pharmacokinetic properties of the aptamer-siRNA chimeras were subsequently measured in vitro and in vivo.

Results

In vitro assays show that albumin-binding aptamers are stable in serum while maintaining potent gene knockdown capabilities in the chimera format. In vivo, the absolute circulation half-life of the best-performing aptamer-siRNA chimera (Clone 1) was 1.6-fold higher than a scrambled aptamer chimera control.

Conclusions

Aptamer-siRNA chimeras exhibit improved bioavailability without compromising biological activity. Hence, this albumin-binding aptamer-siRNA chimera approach may be a promising strategy for drug delivery applications.

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Acknowledgments

Funding for this work was provided by the Chan Zuckerberg Initiative (Grant No. 2018-191850 to ESL), the BrightFocus Foundation (Grant No. A20170945 to ESL), and National Institutes of Health Grants R01 CA224241 and R01 EB019409 (to CLD). ENH was supported by a National Science Foundation Graduate Research Fellowship.

Author Contributions

JCR, ENH, CLD, and ESL conceived the research plan. JCR, ENH, and AGS carried out experiments. All authors wrote, reviewed, and edited the manuscript.

Conflict of interest

The authors have no competing financial interests.

Author information

Author notes

  1. Jonah C. Rosch and Ella N. Hoogenboezem are co-first author.

Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, TN, USA

    Jonah C. Rosch & Ethan S. Lippmann

  2. Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, USA

    Ella N. Hoogenboezem, Alexander G. Sorets, Craig L. Duvall & Ethan S. Lippmann

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  1. Jonah C. Rosch
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  2. Ella N. Hoogenboezem
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  3. Alexander G. Sorets
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  4. Craig L. Duvall
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  5. Ethan S. Lippmann
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Corresponding authors

Correspondence to Craig L. Duvall or Ethan S. Lippmann.

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Associate Editor Michael R. King oversaw the review of this article.

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Rosch, J.C., Hoogenboezem, E.N., Sorets, A.G. et al. Albumin-Binding Aptamer Chimeras for Improved siRNA Bioavailability. Cel. Mol. Bioeng. 15, 161–173 (2022). https://doi.org/10.1007/s12195-022-00718-y

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  • DOI: https://doi.org/10.1007/s12195-022-00718-y

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