DNA topoisomerase inhibition with the HIF inhibitor acriflavine promotes transcription of lncRNAs in endothelial cells

Molecular Therapy: Nucleic Acids

Frederike Boos¹  Stefan Knapp¹  James A. Oo¹  Felix F. Lillich¹  Ralf Gilsbach²  Stefan Günther²  Judit Izquierdo Ponce²  Ewgenij Proschak²  Sandra Seredinski²  Ralf P. Brandes³  Beatrice Pflüger-Müller³  Matthias S. Leisegang³  Timothy Warwick⁴ 

[1] German Center of Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt, Germany;Institute for Cardiovascular Physiology, Goethe University, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany;Institute of Pharmaceutical Chemistry, Goethe University, 60438 Frankfurt, Germany;Max Planck Institute for Heart and Lung Research, 61231 Bad Nauheim, Germany;

关键词: long non-coding RNA;    endothelial cells;    angiogenesis;    chromatin;    topoisomerase inhibitor;    hypoxia;   

DOI  :  

来源: DOAJ

【 摘 要 】

The transcription factor hypoxia-inducible factor 1 (HIF1) is an important driver of cancer and is therefore an attractive drug target. Acriflavine (ACF) has been suggested to inhibit HIF1, but its mechanism of action is unknown. Here we investigated the interaction of ACF with DNA and long non-coding RNAs (lncRNAs) and its function in human endothelial cells. ACF promoted apoptosis and reduced proliferation, network formation, and angiogenic capacity. It also induced changes in gene expression, as determined by RNA sequencing (RNA-seq), which could not be attributed to specific inhibition of HIF1. A similar response was observed in murine lung endothelial cells. Although ACF increased and decreased a similar number of protein-coding genes, lncRNAs were preferentially upregulated under normoxic and hypoxic conditions. An assay for transposase accessibility with subsequent DNA sequencing (ATAC-seq) demonstrated that ACF induced strong changes in chromatin accessibility at lncRNA promoters. Immunofluorescence showed displacement of DNA:RNA hybrids. Such effects might be due to ACF-mediated topoisomerase inhibition, which was indeed the case, as reflected by DNA unwinding assays. Comparison with other acridine derivatives and topoisomerase inhibitors suggested that the specific function of ACF is an effect of acridinium-class compounds. This study demonstrates that ACF inhibits topoisomerases rather than HIF specifically and that it elicits a unique expression response of lncRNAs.

【 授权许可】

Unknown