TY - JOUR
T1 - A Modular Albumin-Oligonucleotide Biomolecular Assembly for Delivery of Antisense Therapeutics
T2 - Molecular Pharmaceutics
AU - Elkhashab, Marwa
AU - Dilek, Yeter
AU - Foss, Morten
AU - Creemers, Laura B.
AU - Howard, Kenneth A.
PY - 2024/2
Y1 - 2024/2
N2 - 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.
AB - 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.
KW - Antisense oligonucleotide
KW - Albumin
KW - Biomolecular assembly
KW - FcRn
KW - ADAMTS5
KW - osteoarthritis
KW - biomolecular assembly, FcRn
KW - antisense oligonucleotide
KW - albumin
KW - Oligonucleotides, Antisense/chemistry
KW - Humans
KW - Oligonucleotides/chemistry
KW - Deoxyribonucleases
KW - Albumins
KW - Endothelial Cells/metabolism
KW - Serum Albumin, Human/metabolism
KW - Osteoarthritis
U2 - 10.1021/acs.molpharmaceut.3c00561
DO - 10.1021/acs.molpharmaceut.3c00561
M3 - Journal article
C2 - 38214218
SN - 1543-8384
VL - 21
SP - 491
EP - 500
JO - Molecular Pharmaceutics
JF - Molecular Pharmaceutics
IS - 2
ER -