A Modular Albumin-Oligonucleotide Biomolecular Assembly for Delivery of Antisense Therapeutics: Molecular Pharmaceutics

Marwa Elkhashab, Yeter Dilek, Morten Foss, Laura B. Creemers, Kenneth A. Howard*

*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

Antisense nucleic acid drugs are susceptible to nuclease degradation, rapid renal clearance, and short circulatory half-life. In this work, we introduce a modular-based recombinant human albumin-oligonucleotide (rHA-cODN) biomolecular assembly that allows incorporation of a chemically stabilized therapeutic gapmer antisense oligonucleotide (ASO) and FcRn-driven endothelial cellular recycling. A phosphodiester ODN linker (cODN) was conjugated to recombinant human albumin (rHA) using maleimide chemistry, after which a complementary gapmer ASO, targeting ADAMTS5 involved in osteoarthritis pathogenesis, was annealed. The rHA-cODN/ASO biomolecular assembly production, fluorescence labeling, and purity were confirmed using polyacrylamide gel electrophoresis. ASO release was triggered by DNase-mediated degradation of the linker strand, reaching 40% in serum after 72 h, with complete release observed following 30 min of incubation with DNase. Cellular internalization and trafficking of the biomolecular assembly using confocal microscopy in C28/I2 cells showed higher uptake and endosomal localization by increasing incubation time from 4 to 24 h. FcRn-mediated cellular recycling of the assembly was demonstrated in FcRn-expressing human microvascular endothelial cells. ADAMTS5 in vitro silencing efficiency reached 40%, which was comparable to free gapmer after 72 h incubation with human osteoarthritis patients’ chondrocytes. This work introduces a versatile biomolecular modular-based “Plug-and-Play” platform potentially applicable for albumin-mediated half-life extension for a range of different types of ODN therapeutics.
Original languageEnglish
JournalMolecular Pharmaceutics
Volume21
Issue2
Pages (from-to)491-500
Number of pages10
ISSN1543-8384
DOIs
Publication statusPublished - Feb 2024

Keywords

  • Antisense oligonucleotide
  • Albumin
  • Biomolecular assembly
  • FcRn
  • ADAMTS5
  • osteoarthritis
  • biomolecular assembly, FcRn
  • antisense oligonucleotide
  • albumin
  • Oligonucleotides, Antisense/chemistry
  • Humans
  • Oligonucleotides/chemistry
  • Deoxyribonucleases
  • Albumins
  • Endothelial Cells/metabolism
  • Serum Albumin, Human/metabolism
  • Osteoarthritis

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