Long Noncoding RNA MANTIS Facilitates Endothelial Angiogenic Function.

Abstract

BACKGROUND:

The angiogenic function of endothelial cells is regulated by numerous mechanisms, but the impact of long noncoding RNAs (lncRNAs) has hardly been studied. We set out to identify novel and functionally important endothelial lncRNAs.

METHODS:

Epigenetically controlled lncRNAs in human umbilical vein endothelial cells were searched by exon-array analysis after knockdown of the histone demethylase JARID1B. Molecular mechanisms were investigated by RNA pulldown and immunoprecipitation, mass spectrometry, microarray, several knockdown approaches, CRISPR-Cas9, assay for transposase-accessible chromatin sequencing, and chromatin immunoprecipitation in human umbilical vein endothelial cells. Patient samples from lung and tumors were studied for MANTIS expression.

RESULTS:

A search for epigenetically controlled endothelial lncRNAs yielded lncRNA n342419, here termed MANTIS, as the most strongly regulated lncRNA. Controlled by the histone demethylase JARID1B, MANTIS was downregulated in patients with idiopathic pulmonary arterial hypertension and in rats treated with monocrotaline, whereas it was upregulated in carotid arteries of Macaca fascicularis subjected to atherosclerosis regression diet, and in endothelial cells isolated from human glioblastoma patients. CRISPR/Cas9-mediated deletion or silencing of MANTIS with small interfering RNAs or GapmeRs inhibited angiogenic sprouting and alignment of endothelial cells in response to shear stress. Mechanistically, the nuclear-localized MANTIS lncRNA interacted with BRG1, the catalytic subunit of the switch/sucrose nonfermentable chromatin-remodeling complex. This interaction was required for nucleosome remodeling by keeping the ATPase function of BRG1 active. Thereby, the transcription of key endothelial genes such as SOX18, SMAD6, and COUP-TFII was regulated by ensuring efficient RNA polymerase II machinery binding.

CONCLUSION:

MANTIS is a differentially regulated novel lncRNA facilitating endothelial angiogenic function.

• Authors: Matthias S Leisegang, Christian Fork, Ivana Josipovic, Florian Martin Richter, Jens Preussner, Jiong Hu, Matthew J Miller, Jeremy Epah, Patrick Hofmann, Stefan Günther, Franziska Moll, Chanil Valasarajan, Juliana Heidler, Yuliya Ponomareva, Thomas M Freiman, Lars Maegdefessel, Karl H Plate, Michel Mittelbronn, Shizuka Uchida, Carsten Künne, Konstantinos Stellos, Ralph T Schermuly, Norbert Weissmann, Kavi Devraj, Ilka Wittig, Reinier A Boon, Stefanie Dimmeler, Soni Savai Pullamsetti, Mario Looso, Francis J Miller Jr, Ralf P Brandes
• Journal: Circulation (Circulation) Vol. 136 Issue 1 Pg. 65-79 (Jul 04 2017) ISSN: 1524-4539 [Electronic] United States
• PMID: 28351900 (Publication Type: Journal Article)
• Copyright: © 2017 The Authors.

Chemical References

• Nuclear Proteins
• RNA, Long Noncoding
• Repressor Proteins
• Jumonji Domain-Containing Histone Demethylases
• KDM5B protein, human

Topics

• Animals
• CRISPR-Cas Systems (physiology)
• Cell Line
• Epigenesis, Genetic (physiology)
• Human Umbilical Vein Endothelial Cells (physiology)
• Humans
• Hypertension, Pulmonary (genetics, metabolism)
• Jumonji Domain-Containing Histone Demethylases (biosynthesis, genetics)
• Macaca fascicularis
• Male
• Mice
• Mice, SCID
• Microvessels (physiology)
• Neovascularization, Physiologic (physiology)
• Nuclear Proteins (biosynthesis, genetics)
• RNA, Long Noncoding (biosynthesis, genetics)
• Rats
• Rats, Sprague-Dawley
• Repressor Proteins (biosynthesis, genetics)