Publikationsdatenbank

VIM, TPI and MAT2A - pivotal proteins for angiogenesis in vitro? (2022)

Art

Vortrag

Autoren

Herre, Christina (WE 1)

Nshdejan, Arpenik (WE 1)

Klopfleisch, Robert (WE 12)

Corte, Giuliano Mario (WE 1)

Bahramsoltani, Mahtab (WE 1)

Kongress

11th Meeting of the Young Generation of Veterinary Anatomists

Zurich, Switzerland, 20. – 22.07.2022

Quelle

Sprache

Englisch

Kontakt

Institut für Veterinär-Anatomie

Koserstr. 20
14195 Berlin
+49 30 838 75784
anatomie@vetmed.fu-berlin.de

Abstract / Zusammenfassung

The ability of endothelial cells (ECs) to undergo angiogenesis is indispensable for many pathological events, e.g. ischemic diseases, tumorigenesis and cardiovascular diseases. Current efforts in the research field of angiogenesis are focusing on tissue engineering and wound healing. For in vitro assays of angiogenesis, reproducibility is still an issue of concern, allegedly based on the heterogenous character of ECs. Being classified into angiogenic and non-angiogenic ECs, it was shown that both groups display differences in their expression of eight specific proteins (1). This study aims to analyze three amongst these proteins, i.e. vimentin (VIM), triosephosphate isomerase (TPI) and adenosylmethionine synthetase isoform type-2 (MAT2A), and their association to angiogenesis in vitro. Therefore, the distribution of tip cells and stalk cells within cell populations of human dermal microvascular endothelial cells was characterized via VEGFR-1 and VEGFR-2 expression. Cells were long-term cultivated and stimulated to undergo angiogenesis via a proangiogenic media. Protein and mRNA expression patterns of all proteins were detected while ECs were running through the angiogenic cascade. According to the all-in-one assay, quantification of angiogenesis was performed by staging the time-dependent morphological changes of cells (2). Furthermore, a shRNA mediated knockdown of VIM was performed and its impact on angiogenesis in vitro and on TPI and MAT2A expression was examined. Opposed to native ECs, knockdown cells were unable to enter late stages of angiogenesis and eventually died. Exclusively cells with higher VEGFR-1 expression survived and displayed an increase in MAT2A and TPI expression. In native cells, VIM and MAT2A were intensely expressed in early angiogenic stages and TPI during the course of angiogenesis in vitro. In conclusion, VIM knockdown led to deceleration of angiogenesis in vitro, cell death of populations with less stalk cells and expressional alterations of TPI and MAT2A. Additionally, native cells provided an indication of VIM and MAT2A being relevant in beginning stages of angiogenesis and TPI during the whole angiogenic cascade in vitro.

[1] Bahramsoltani M, Harms T, Drewes B, Plendl J (2013) Searching for markers to identify angiogenic endothelial cells: a proteomic approach, Clinical Hemorheology and Microcirculation:55, 255-69.
[2] Bahramsoltani M, Spiegelaere W de, Janczyk P, Hiebl B, Cornillie P, Plendl J (2010) Quantitation of angiogenesis in vitro induced by VEGF-A and FGF-2 in two different human endothelial cultures – an all-in-one assay, Clinical Hemorheology and Microcirculation:46, 189-202.