Endosomal Escape of Antisense Oligonucleotides Internalized by Stabilin Receptors Is Regulated by Rab5C and EEA1 during Endosomal Maturation

Colton M. Miller, W. Brad Wan, Punit P. Seth, Edward N Harris

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Second-generation (Gen 2) Antisense oligonucleotides (ASOs) show increased nuclease stability and affinity for their RNA targets, which has translated to improved potency and therapeutic index in the clinic. Gen 2 ASOs are typically modified using the phosphorothioate (PS) backbone modification, which enhances ASO interactions with plasma, cell surface, and intracellular proteins. This facilitates ASO distribution to peripheral tissues and also promotes cellular uptake after injection into animals. Previous work identified that Stabilin receptors specifically internalize PS-ASOs in the sinusoidal endothelial cells of the liver and the spleen. By modulating expression of specific proteins involved in the trafficking and maturation of the endolysosomal compartments, we show that Rab5C and EEA1 in the early endosomal pathway, and Rab7A and lysobisphosphatidic acid in the late endosomal pathway, are important for trafficking of PS-ASOs and facilitate their escape from endolysosomal compartments after Stabilin-mediated internalization. In conclusion, this work identifies key rate-limiting proteins in the pathway for PS-ASO translocation and escape from the endosome.

Original languageEnglish (US)
Pages (from-to)86-96
Number of pages11
JournalNucleic Acid Therapeutics
Volume28
Issue number2
DOIs
StatePublished - Apr 1 2018

Fingerprint

Antisense Oligonucleotides
Phosphorothioate Oligonucleotides
Proteins
Endosomes
Endothelial cells
Beam plasma interactions
Plasma Cells
Liver
Membrane Proteins
Animals
Spleen
Endothelial Cells
RNA
Tissue
Plasmas
Injections

Keywords

  • ASO
  • Stabilin
  • antisense oligonucleotide
  • endosome escape
  • endosome maturation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Drug Discovery

Cite this

Endosomal Escape of Antisense Oligonucleotides Internalized by Stabilin Receptors Is Regulated by Rab5C and EEA1 during Endosomal Maturation. / Miller, Colton M.; Wan, W. Brad; Seth, Punit P.; Harris, Edward N.

In: Nucleic Acid Therapeutics, Vol. 28, No. 2, 01.04.2018, p. 86-96.

Research output: Contribution to journalArticle

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