Self-assembly into nanoparticles is essential for receptor mediated uptake of therapeutic antisense oligonucleotides

Ahmed, Kariem; Aoki, Yoshitsugu; Koo, Taeyoung; McClorey, Graham; Benner, Leif; Coenen-Stass, Anna; O'Donovan, Liz; Lehto, Taavi; Garciaguerra, Antonio; Nordin, Joel Zacharias; Saleh, Amer F.; Behlke, Mark A.; Morris, John; Goyenvalle, Aurelie; Dugovic, Branislav; Leumann, Christian Joerg; Gordon, Siamon; Gait, Michael J.; El-Andaloussi, Samir and Wood, Matthew J. A. (2015). Self-assembly into nanoparticles is essential for receptor mediated uptake of therapeutic antisense oligonucleotides. Nano letters, 15(7), pp. 4364-4374. American Chemical Society 10.1021/acs.nanolett.5b00490

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Antisense oligonucleotides (ASOs) have the potential of revolutionizing medicine due to their ability to manipulate gene function for therapeutic purposes. ASOs are chemically modified and/or incorporated with nanoparticles to enhance their stability and cellular uptake; however, one of the biggest challenges is the poor understanding of their uptake mechanism, which is needed for designing better ASOs with high activity and low toxicity. Here, we study the uptake mechanism of three therapeutically relevant ASOs (peptide-conjugated phosphorodiamidate morpholino (P-PMO), 2?Omethyl phosphorothioate (2?OMe) and phosphorothioated tricyclo DNA (tcDNA) that have been optimized to induce exon skipping in models of Deuchenne muscular dystrophy (DMD). We show that P-PMO and tcDNA have high propensity to spontaneously self-assemble into nanoparticles. P-PMO forms micelles of defined size and their net charge (zeta potential) is dependent on the medium and concentration. In biomimetic conditions and at low concentrations P-PMO obtains net negative charge and its uptake is mediated by class A scavenger receptor subtypes (SCARAs) as shown by competitive inhibition and RNAi silencing experiments in-vitro. In-vivo, the activity of P-PMO was significantly decreased in SCARA1 knock-out mice compared to wild-type animals. Additionally, we show that SCARA1 is involved in the uptake of tcDNA and 2?OMe as shown by competitive inhibition and co-localization experiments. Surface plasmon resonance binding analysis to SCARA1 demonstrated that P-PMO and tcDNA have higher binding profiles to the receptor compared to 2?OMe. These results demonstrate receptor-mediated uptake for a range of ASO chemistries, a mechanism that is dependent on their self-assembly into nanoparticles.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Dugovic, Branislav and Leumann, Christian

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

1530-6984

Publisher:

American Chemical Society

Language:

English

Submitter:

Christian Leumann

Date Deposited:

11 Jun 2015 10:29

Last Modified:

05 Jun 2016 02:30

Publisher DOI:

10.1021/acs.nanolett.5b00490

PubMed ID:

26042553

Additional Information:

Date: 2015/06/04

BORIS DOI:

10.7892/boris.69427

URI:

https://boris.unibe.ch/id/eprint/69427

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