Hydrogel-Assisted Antisense LNA Gapmer Delivery for In Situ Gene Silencing in Spinal Cord Injury

Pedro M.D. Moreno, Ana R. Ferreira, Daniela Salvador, Maria T. Rodrigues, Marília Torrado, Eva D. Carvalho, Ulf Tedebark, Mónica M. Sousa, Isabel F. Amaral, Jesper Wengel, Ana P. Pêgo*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

226 Downloads (Pure)

Abstract

After spinal cord injury (SCI), nerve regeneration is severely hampered due to the establishment of a highly inhibitory microenvironment at the injury site, through the contribution of multiple factors. The potential of antisense oligonucleotides (AONs) to modify gene expression at different levels, allowing the regulation of cell survival and cell function, together with the availability of chemically modified nucleic acids with favorable biopharmaceutical properties, make AONs an attractive tool for novel SCI therapy developments. In this work, we explored the potential of locked nucleic acid (LNA)-modified AON gapmers in combination with a fibrin hydrogel bridging material to induce gene silencing in situ at a SCI lesion site. LNA gapmers were effectively developed against two promising gene targets aiming at enhancing axonal regeneration—RhoA and GSK3β. The fibrin-matrix-assisted AON delivery system mediated potent RNA knockdown in vitro in a dorsal root ganglion explant culture system and in vivo at a SCI lesion site, achieving around 75% downregulation 5 days after hydrogel injection. Our results show that local implantation of a AON-gapmer-loaded hydrogel matrix mediated efficient gene silencing in the lesioned spinal cord and is an innovative platform that can potentially combine gene regulation with regenerative permissive substrates aiming at SCI therapeutics and nerve regeneration.

Original languageEnglish
JournalMolecular Therapy - Nucleic Acids
Volume11
Pages (from-to)393-406
ISSN2162-2531
DOIs
Publication statusPublished - 2018

Fingerprint

Hydrogel
Spinal Ganglia
Nucleic Acids
Cell Survival
Down-Regulation
RNA
Wounds and Injuries

Keywords

  • antisense oligonucleotides
  • gapmer
  • hydrogel
  • in situ delivery
  • LNA
  • spinal cord injury

Cite this

Moreno, P. M. D., Ferreira, A. R., Salvador, D., Rodrigues, M. T., Torrado, M., Carvalho, E. D., ... Pêgo, A. P. (2018). Hydrogel-Assisted Antisense LNA Gapmer Delivery for In Situ Gene Silencing in Spinal Cord Injury. Molecular Therapy - Nucleic Acids, 11, 393-406. https://doi.org/10.1016/j.omtn.2018.03.009
Moreno, Pedro M.D. ; Ferreira, Ana R. ; Salvador, Daniela ; Rodrigues, Maria T. ; Torrado, Marília ; Carvalho, Eva D. ; Tedebark, Ulf ; Sousa, Mónica M. ; Amaral, Isabel F. ; Wengel, Jesper ; Pêgo, Ana P. / Hydrogel-Assisted Antisense LNA Gapmer Delivery for In Situ Gene Silencing in Spinal Cord Injury. In: Molecular Therapy - Nucleic Acids. 2018 ; Vol. 11. pp. 393-406.
@article{8b3146feb1cf4beaaaf601772cafa8a6,
title = "Hydrogel-Assisted Antisense LNA Gapmer Delivery for In Situ Gene Silencing in Spinal Cord Injury",
abstract = "After spinal cord injury (SCI), nerve regeneration is severely hampered due to the establishment of a highly inhibitory microenvironment at the injury site, through the contribution of multiple factors. The potential of antisense oligonucleotides (AONs) to modify gene expression at different levels, allowing the regulation of cell survival and cell function, together with the availability of chemically modified nucleic acids with favorable biopharmaceutical properties, make AONs an attractive tool for novel SCI therapy developments. In this work, we explored the potential of locked nucleic acid (LNA)-modified AON gapmers in combination with a fibrin hydrogel bridging material to induce gene silencing in situ at a SCI lesion site. LNA gapmers were effectively developed against two promising gene targets aiming at enhancing axonal regeneration—RhoA and GSK3β. The fibrin-matrix-assisted AON delivery system mediated potent RNA knockdown in vitro in a dorsal root ganglion explant culture system and in vivo at a SCI lesion site, achieving around 75{\%} downregulation 5 days after hydrogel injection. Our results show that local implantation of a AON-gapmer-loaded hydrogel matrix mediated efficient gene silencing in the lesioned spinal cord and is an innovative platform that can potentially combine gene regulation with regenerative permissive substrates aiming at SCI therapeutics and nerve regeneration.",
keywords = "antisense oligonucleotides, gapmer, hydrogel, in situ delivery, LNA, spinal cord injury",
author = "Moreno, {Pedro M.D.} and Ferreira, {Ana R.} and Daniela Salvador and Rodrigues, {Maria T.} and Mar{\'i}lia Torrado and Carvalho, {Eva D.} and Ulf Tedebark and Sousa, {M{\'o}nica M.} and Amaral, {Isabel F.} and Jesper Wengel and P{\^e}go, {Ana P.}",
year = "2018",
doi = "10.1016/j.omtn.2018.03.009",
language = "English",
volume = "11",
pages = "393--406",
journal = "Molecular Therapy - Nucleic Acids",
issn = "2162-2531",
publisher = "Nature Publishing Group",

}

Moreno, PMD, Ferreira, AR, Salvador, D, Rodrigues, MT, Torrado, M, Carvalho, ED, Tedebark, U, Sousa, MM, Amaral, IF, Wengel, J & Pêgo, AP 2018, 'Hydrogel-Assisted Antisense LNA Gapmer Delivery for In Situ Gene Silencing in Spinal Cord Injury', Molecular Therapy - Nucleic Acids, vol. 11, pp. 393-406. https://doi.org/10.1016/j.omtn.2018.03.009

Hydrogel-Assisted Antisense LNA Gapmer Delivery for In Situ Gene Silencing in Spinal Cord Injury. / Moreno, Pedro M.D.; Ferreira, Ana R.; Salvador, Daniela; Rodrigues, Maria T.; Torrado, Marília; Carvalho, Eva D.; Tedebark, Ulf; Sousa, Mónica M.; Amaral, Isabel F.; Wengel, Jesper; Pêgo, Ana P.

In: Molecular Therapy - Nucleic Acids, Vol. 11, 2018, p. 393-406.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Hydrogel-Assisted Antisense LNA Gapmer Delivery for In Situ Gene Silencing in Spinal Cord Injury

AU - Moreno, Pedro M.D.

AU - Ferreira, Ana R.

AU - Salvador, Daniela

AU - Rodrigues, Maria T.

AU - Torrado, Marília

AU - Carvalho, Eva D.

AU - Tedebark, Ulf

AU - Sousa, Mónica M.

AU - Amaral, Isabel F.

AU - Wengel, Jesper

AU - Pêgo, Ana P.

PY - 2018

Y1 - 2018

N2 - After spinal cord injury (SCI), nerve regeneration is severely hampered due to the establishment of a highly inhibitory microenvironment at the injury site, through the contribution of multiple factors. The potential of antisense oligonucleotides (AONs) to modify gene expression at different levels, allowing the regulation of cell survival and cell function, together with the availability of chemically modified nucleic acids with favorable biopharmaceutical properties, make AONs an attractive tool for novel SCI therapy developments. In this work, we explored the potential of locked nucleic acid (LNA)-modified AON gapmers in combination with a fibrin hydrogel bridging material to induce gene silencing in situ at a SCI lesion site. LNA gapmers were effectively developed against two promising gene targets aiming at enhancing axonal regeneration—RhoA and GSK3β. The fibrin-matrix-assisted AON delivery system mediated potent RNA knockdown in vitro in a dorsal root ganglion explant culture system and in vivo at a SCI lesion site, achieving around 75% downregulation 5 days after hydrogel injection. Our results show that local implantation of a AON-gapmer-loaded hydrogel matrix mediated efficient gene silencing in the lesioned spinal cord and is an innovative platform that can potentially combine gene regulation with regenerative permissive substrates aiming at SCI therapeutics and nerve regeneration.

AB - After spinal cord injury (SCI), nerve regeneration is severely hampered due to the establishment of a highly inhibitory microenvironment at the injury site, through the contribution of multiple factors. The potential of antisense oligonucleotides (AONs) to modify gene expression at different levels, allowing the regulation of cell survival and cell function, together with the availability of chemically modified nucleic acids with favorable biopharmaceutical properties, make AONs an attractive tool for novel SCI therapy developments. In this work, we explored the potential of locked nucleic acid (LNA)-modified AON gapmers in combination with a fibrin hydrogel bridging material to induce gene silencing in situ at a SCI lesion site. LNA gapmers were effectively developed against two promising gene targets aiming at enhancing axonal regeneration—RhoA and GSK3β. The fibrin-matrix-assisted AON delivery system mediated potent RNA knockdown in vitro in a dorsal root ganglion explant culture system and in vivo at a SCI lesion site, achieving around 75% downregulation 5 days after hydrogel injection. Our results show that local implantation of a AON-gapmer-loaded hydrogel matrix mediated efficient gene silencing in the lesioned spinal cord and is an innovative platform that can potentially combine gene regulation with regenerative permissive substrates aiming at SCI therapeutics and nerve regeneration.

KW - antisense oligonucleotides

KW - gapmer

KW - hydrogel

KW - in situ delivery

KW - LNA

KW - spinal cord injury

U2 - 10.1016/j.omtn.2018.03.009

DO - 10.1016/j.omtn.2018.03.009

M3 - Journal article

C2 - 29858074

AN - SCOPUS:85045213607

VL - 11

SP - 393

EP - 406

JO - Molecular Therapy - Nucleic Acids

JF - Molecular Therapy - Nucleic Acids

SN - 2162-2531

ER -