Nuclear envelope morphology change upon repetitive treatment with modified antisense oligonucleotides targeting Hutchinson-Gilford Progeria Syndrome

Asmaa Abdelrahman; Mette-Marie Wendelboe Nielsen; Mette Halkjær Stage; Eva Christensen Arnspang

doi:10.1016/j.bbrep.2022.101411

Nuclear envelope morphology change upon repetitive treatment with modified antisense oligonucleotides targeting Hutchinson-Gilford Progeria Syndrome

Biochem Biophys Rep. 2022 Dec 28:33:101411. doi: 10.1016/j.bbrep.2022.101411. eCollection 2023 Mar.

Authors

Asmaa Abdelrahman^{1

2}, Mette-Marie Wendelboe Nielsen^{1

3}, Mette Halkjær Stage^{1

4}, Eva Christensen Arnspang¹

Affiliations

¹ Department of Green Technology, Faculty of Engineering, University of Southern Denmark, Odense, Denmark.
² Department of Photochemistry, National Research Centre, Dokki, Giza, Egypt.
³ Department of Mechanical and Electrical Engineering, Faculty of Engineering University of Southern Denmark, Sønderborg, Denmark.
⁴ Department of Food Science, Faculty of Science, Copenhagen University, Copenhagen, Denmark.

Abstract

We present the influence of treating progeroid fibroblasts with two modified antisense oligonucleotides (ONs) on the nuclear envelope. Two modified ONs were designed to block ribosome binding during translation and spliceosome binding at the cryptic splice site. We analysed the changes in the nuclear morphology of progeria cell nuclei after repetitive transfection with modified ONs as a physical analysis tool for estimating alteration of the gene expression at the protein level. Confocal microscopy was used to image the nuclei, and the nuclear lobulations were quantified to study the changes in the morphology of the nuclear envelope upon treatment. PCR was used to identify the changes in the expression of lamin A and progerin after antisense treatment at the RNA level. We found a significant decrease in the number of nuclear envelope lobulations and a lower progerin expression in progeria cells after transfection with modified ONs.

Keywords: Confocal microscopy; Gene silencing; Genetic mutation; Rare disease.