Stabilin-1 and Stabilin-2 are specific receptors for the cellular internalization of phosphorothioate-modified antisense oligonucleotides (ASOs) in the liver

Colton M. Miller, Aaron J. Donner, Emma E. Blank, Andrew W. Egger, Brianna M. Kellar, Michael E. Østergaard, Punit P. Seth, Edward N. Harris

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Phosphorothioate (PS)-modified antisense oligonucleotides (ASOs) have been extensively investigated over the past three decades as pharmacological and therapeutic agents. One second generation ASO, Kynamro™, was recently approved by the FDA for the treatment of homozygous familial hypercholesterolemia and over 35 second generation PS ASOs are at various stages of clinical development. In this report, we show that the Stabilin class of scavenger receptors, which were not previously thought to bind DNA, do bind and internalize PS ASOs. With the use of primary cells from mouse and rat livers and recombinant cell lines each expressing Stabilin-1 and each isoform of Stabilin-2 (315-HARE and 190-HARE), we have determined that PS ASOs bind with high affinity and these receptors are responsible for bulk, clathrin-mediated endocytosis within the cell. Binding is primarily dependent on salt-bridge formation and correct folding of the intact protein receptor. Increased internalization rates also enhanced ASO potency for reducing expression of the non-coding RNA Malat-1, in Stabilin-expressing cell lines. A more thorough understanding of mechanisms by which ASOs are internalized in cells and their intracellular trafficking pathways will aid in the design of next generation antisense agents with improved therapeutic properties.

Original languageEnglish (US)
Pages (from-to)2782-2794
Number of pages13
JournalNucleic acids research
Volume44
Issue number6
DOIs
StatePublished - Feb 22 2016

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Antisense Oligonucleotides
Phosphorothioate Oligonucleotides
Liver
Scavenger Receptors
Cell Line
Untranslated RNA
Clathrin
Hyperlipoproteinemia Type II
Protein Folding
Endocytosis
Protein Isoforms
Salts
Pharmacology
DNA
Therapeutics

ASJC Scopus subject areas

  • Genetics

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Stabilin-1 and Stabilin-2 are specific receptors for the cellular internalization of phosphorothioate-modified antisense oligonucleotides (ASOs) in the liver. / Miller, Colton M.; Donner, Aaron J.; Blank, Emma E.; Egger, Andrew W.; Kellar, Brianna M.; Østergaard, Michael E.; Seth, Punit P.; Harris, Edward N.

In: Nucleic acids research, Vol. 44, No. 6, 22.02.2016, p. 2782-2794.

Research output: Contribution to journalArticle

Miller, Colton M. ; Donner, Aaron J. ; Blank, Emma E. ; Egger, Andrew W. ; Kellar, Brianna M. ; Østergaard, Michael E. ; Seth, Punit P. ; Harris, Edward N. / Stabilin-1 and Stabilin-2 are specific receptors for the cellular internalization of phosphorothioate-modified antisense oligonucleotides (ASOs) in the liver. In: Nucleic acids research. 2016 ; Vol. 44, No. 6. pp. 2782-2794.
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