Novel endosomolytic compounds enable highly potent delivery of antisense oligonucleotides
Journal article, 2022

The therapeutic and research potentials of oligonucleotides (ONs) have been hampered in part by their inability to effectively escape endosomal compartments to reach their cytosolic and nuclear targets. Splice-switching ONs (SSOs) can be used with endosomolytic small molecule compounds to increase functional delivery. So far, development of these compounds has been hindered by a lack of high-resolution methods that can correlate SSO trafficking with SSO activity. Here we present in-depth characterization of two novel endosomolytic compounds by using a combination of microscopic and functional assays with high spatiotemporal resolution. This system allows the visualization of SSO trafficking, evaluation of endosomal membrane rupture, and quantitates SSO functional activity on a protein level in the presence of endosomolytic compounds. We confirm that the leakage of SSO into the cytosol occurs in parallel with the physical engorgement of LAMP1-positive late endosomes and lysosomes. We conclude that the new compounds interfere with SSO trafficking to the LAMP1-positive endosomal compartments while inducing endosomal membrane rupture and concurrent ON escape into the cytosol. The efficacy of these compounds advocates their use as novel, potent, and quick-acting transfection reagents for antisense ONs.

Author

Jeremy P. Bost

Karolinska Institutet

Miina Ojansivu

Karolinska Institutet

Michael J. Munson

AstraZeneca AB

Emelie Vilhelmsson Wesén

Chalmers, Biology and Biological Engineering, Chemical Biology

Audrey Gallud

Chalmers, Biology and Biological Engineering, Chemical Biology

AstraZeneca AB

Dhanu Gupta

Karolinska Institutet

Oskar Gustafsson

Karolinska Institutet

Osama Saher

Karolinska Institutet

Cairo University

Julia Rädler

Karolinska Institutet

Stuart G. Higgins

Imperial College London

Taavi Lehto

University of Tartu

Karolinska Institutet

Margaret N. Holme

Karolinska Institutet

Anders Dahlén

AstraZeneca AB

Ola Engkvist

AstraZeneca AB

Per Erik Strömstedt

AstraZeneca AB

Shalini Andersson

AstraZeneca AB

C. I. Edvard Smith

Karolinska Institutet

Molly M. Stevens

Karolinska Institutet

Imperial College London

Elin Esbjörner Winters

Chalmers, Biology and Biological Engineering, Chemical Biology

Anna Collén

AstraZeneca AB

Samir El-Andaloussi

Karolinska Institutet

Communications Biology

23993642 (eISSN)

Vol. 5 1 185

Funktionell leverans av nukleotid-baserade läkemedel

Swedish Foundation for Strategic Research (SSF) (IRC15-0065), 2017-03-01 -- 2024-12-31.

Subject Categories

Biochemistry and Molecular Biology

Medicinal Chemistry

Organic Chemistry

DOI

10.1038/s42003-022-03132-2

PubMed

35233031

More information

Latest update

3/14/2022