Role of long non-coding RNA CRNDE in the pathophysiology of thoracic aortic aneurysm - an in vitro analysis.
Final Report Abstract
As part of the DFG Young Investigators Academy in Cardiac Surgery, this project focused on the study of the long non-coding RNA (lncRNA) CRNDE (Colorectal Neoplasi-a Differentially Expressed) in thoracic aortic aneurysms (TAA). CRNDE is a lncRNA that is selectively expressed in vascular smooth muscle cells (VSMC) in abdominal aortic aneurysms. Quantitative expression analysis by qPCR, fluorescence microscopy localization determinations and functional analysis using life cell imaging and a 3D cell culture model was planned. Furthermore, we aimed to distinguish dilated and non-dilated TAA regions. In qPCR, there was no significant difference between dilated and non-dilated tissues in TAA samples. Therefore, the original plan was abandoned and scRNAseq of 6 tissue samples each in dilated and nondilated areas in TAAs was performed to identify relevant lncRNAs. In the analysis, the lncRNAs LINC00632 and TMEM72-AS1 showed almost isolated expression in the VSMC clusters with significantly higher expression in the dilated region. Quantitatively, this was reflected in qPCR. Both lncRNAs showed significantly increased expression in dilated regions. Functional analyses were performed on isolated VSMCs from TAA tissues. Using siRNAs, temporary cellular knock-out (KO) of TMEM72-AS1 and LINC00632 was performed. The effect of the KO cell model on proliferation, migration, and apoptosis was then examined by life cell imaging. Both LINC00632 and TMEM72-AS1 inhibit migration and proliferation. Furthermore, both lncRNAs activate apoptosis. These findings suggest that both TMEM72-AS1 and LINC00632 play a relevant role in the pathophysiology of thoracic aortic aneurysm.
