ADAMTS5-specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasound

Objective Antisense oligonucleotides (ASOs) have reached the clinic; however, they lack tissue specificity. Albumin is a plasma-abundant macromolecule that has been shown to accumulate in inflamed tissues. In this work, we have designed a recombinant human albumin (rHA)-based biomolecular assembly i...

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Main Authors: Marwa Elkhashab, Goncalo Barreto, Maxime Fauconnier, Yohann Le Bourlout, Laura B. Creemers, Heikki J. Nieminen, Kenneth A. Howard
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:Drug Delivery
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Online Access:https://www.tandfonline.com/doi/10.1080/10717544.2025.2464921
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author Marwa Elkhashab
Goncalo Barreto
Maxime Fauconnier
Yohann Le Bourlout
Laura B. Creemers
Heikki J. Nieminen
Kenneth A. Howard
author_facet Marwa Elkhashab
Goncalo Barreto
Maxime Fauconnier
Yohann Le Bourlout
Laura B. Creemers
Heikki J. Nieminen
Kenneth A. Howard
author_sort Marwa Elkhashab
collection DOAJ
description Objective Antisense oligonucleotides (ASOs) have reached the clinic; however, they lack tissue specificity. Albumin is a plasma-abundant macromolecule that has been shown to accumulate in inflamed tissues. In this work, we have designed a recombinant human albumin (rHA)-based biomolecular assembly incorporating a DNase-resistant phosphorothioate-based complementary oligonucleotide (cODN) and an anti-ADAMTS5 ASO for potential delivery to inflamed sites. Ultrasound (US) was used to trigger ASO release from the assembly and enhance internalization into articular cartilage.Methods A phosphorothioate cODN was conjugated to rHA through a maleimide cross-linker after which, a therapeutic ADAMTS5-specific gapmer ASO was annealed to the cODN. ASO release was assessed after exposing the biomolecular assembly to different US conditions using an US-actuated medical needle operating at 32.2 kHz. Gene silencing efficiency of US-treated anti-ADAMTS5 ASO was assessed in human primary chondrocytes isolated from osteoarthritic patients. US-mediated ASO penetration into articular cartilage was assessed on ex vivo bovine articular cartilage.Results ASO release was observed after exposure to US waves in continuous mode conditions that did not compromise ASO gene silencing efficiency in human chondrocytes. Furthermore, US increased ASO internalization into bovine articular cartilage after 30 min of application without detrimental effects on chondrocyte viability.Conclusion A medical needle driven by continuous US waves at 32.2 kHz has the capability of disassembling a duplex oligonucleotide and enhancing released ASOs internalization into articular cartilage. This work offers the potential delivery and the local triggered release of ASOs at the surface of articular cartilage providing potential benefits for the treatment of diverse cartilage pathologies.
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spelling doaj-art-b41380ebc6d348dabcef359ecc14a3392025-08-20T02:29:59ZengTaylor & Francis GroupDrug Delivery1071-75441521-04642025-12-0132110.1080/10717544.2025.2464921ADAMTS5-specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasoundMarwa Elkhashab0Goncalo Barreto1Maxime Fauconnier2Yohann Le Bourlout3Laura B. Creemers4Heikki J. Nieminen5Kenneth A. Howard6Interdisciplinary Nanoscience Center (iNANO) and Department of Molecular Biology, Aarhus University, Aarhus C, DenmarkMedical Ultrasonics Laboratory (MEDUSA), Department of Neuroscience and Biomedical Engineering, Aalto University, Espoo, FinlandMedical Ultrasonics Laboratory (MEDUSA), Department of Neuroscience and Biomedical Engineering, Aalto University, Espoo, FinlandMedical Ultrasonics Laboratory (MEDUSA), Department of Neuroscience and Biomedical Engineering, Aalto University, Espoo, FinlandDepartment of Orthopedics, University Medical Center Utrecht, Utrecht, The NetherlandsMedical Ultrasonics Laboratory (MEDUSA), Department of Neuroscience and Biomedical Engineering, Aalto University, Espoo, FinlandInterdisciplinary Nanoscience Center (iNANO) and Department of Molecular Biology, Aarhus University, Aarhus C, DenmarkObjective Antisense oligonucleotides (ASOs) have reached the clinic; however, they lack tissue specificity. Albumin is a plasma-abundant macromolecule that has been shown to accumulate in inflamed tissues. In this work, we have designed a recombinant human albumin (rHA)-based biomolecular assembly incorporating a DNase-resistant phosphorothioate-based complementary oligonucleotide (cODN) and an anti-ADAMTS5 ASO for potential delivery to inflamed sites. Ultrasound (US) was used to trigger ASO release from the assembly and enhance internalization into articular cartilage.Methods A phosphorothioate cODN was conjugated to rHA through a maleimide cross-linker after which, a therapeutic ADAMTS5-specific gapmer ASO was annealed to the cODN. ASO release was assessed after exposing the biomolecular assembly to different US conditions using an US-actuated medical needle operating at 32.2 kHz. Gene silencing efficiency of US-treated anti-ADAMTS5 ASO was assessed in human primary chondrocytes isolated from osteoarthritic patients. US-mediated ASO penetration into articular cartilage was assessed on ex vivo bovine articular cartilage.Results ASO release was observed after exposure to US waves in continuous mode conditions that did not compromise ASO gene silencing efficiency in human chondrocytes. Furthermore, US increased ASO internalization into bovine articular cartilage after 30 min of application without detrimental effects on chondrocyte viability.Conclusion A medical needle driven by continuous US waves at 32.2 kHz has the capability of disassembling a duplex oligonucleotide and enhancing released ASOs internalization into articular cartilage. This work offers the potential delivery and the local triggered release of ASOs at the surface of articular cartilage providing potential benefits for the treatment of diverse cartilage pathologies.https://www.tandfonline.com/doi/10.1080/10717544.2025.2464921Albuminbiomolecular assemblyAntisenseultrasoundcartilage
spellingShingle Marwa Elkhashab
Goncalo Barreto
Maxime Fauconnier
Yohann Le Bourlout
Laura B. Creemers
Heikki J. Nieminen
Kenneth A. Howard
ADAMTS5-specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasound
Drug Delivery
Albumin
biomolecular assembly
Antisense
ultrasound
cartilage
title ADAMTS5-specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasound
title_full ADAMTS5-specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasound
title_fullStr ADAMTS5-specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasound
title_full_unstemmed ADAMTS5-specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasound
title_short ADAMTS5-specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasound
title_sort adamts5 specific gapmer release from an albumin biomolecular assembly and cartilage internalization triggered by ultrasound
topic Albumin
biomolecular assembly
Antisense
ultrasound
cartilage
url https://www.tandfonline.com/doi/10.1080/10717544.2025.2464921
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