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 185Funktionell 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