TY - JOUR
T1 - Transcriptional and functional profiling identifies inflammation and endothelial-to-mesenchymal transition as potential drivers for phenotypic heterogeneity within a cohort of endothelial colony forming cells
AU - Laan, Sebastiaan N.J.
AU - de Boer, Suzan
AU - SYMPHONY consortium
AU - Dirven, Richard J.
AU - van Moort, Iris
AU - Kuipers, Thomas B.
AU - Mei, Hailiang
AU - Bierings, Ruben
AU - Eikenboom, Jeroen
AU - Brands, Martijn
AU - Koopman, Sjoerd
AU - Bukkems, Laura
AU - Cloesmeijer, Michael
AU - Janssen, Alexander
AU - Fijnvandraat, Karin
AU - Gouw, Samantha
AU - Mathôt, Ron
AU - Haverman, Lotte
AU - van den Akker, Emile
AU - van den Biggelaar, Maartje
AU - de Haas, Masja
AU - Meijer, Sander
AU - Voorberg, Jan
AU - Del Castillo Alferez, Jessica
AU - Zhang, Huan
AU - Boender, Johan
AU - Meijer, Stephan
AU - Meijer, Karina
AU - de Jong, Sean
AU - Laan, Sebastiaan N.J.
AU - Schols, Saskia
AU - Burdorf, Lex
AU - Cnossen, Marjon
AU - Hazelzet, Jan
AU - Huisman, Elise
AU - Kruip, Marieke
AU - Leebeek, Frank
AU - van Leeuwen, Nikki
AU - Lingsma, Hester
AU - de Maat, Moniek
AU - Polinder, Suzanne
AU - Reitsma, Simone
AU - Arisz, Ryanne
AU - Romano, Lorenzo
AU - Al Arashi, Wala
AU - van Hoorn, Shannon
AU - Goedhart, Tine
AU - Uyl, Carin
AU - Jansen, Nathalie
AU - Zivkovic, Minka
AU - Bredenoord, Annelien
N1 - Publisher Copyright:
© 2024 The Author(s)
PY - 2024/7
Y1 - 2024/7
N2 - Background: Endothelial colony-forming cells (ECFCs) derived from patients can be used to investigate pathogenic mechanisms of vascular diseases like von Willebrand disease. Considerable phenotypic heterogeneity has been observed between ECFC clones derived from healthy donors. This heterogeneity needs to be well understood in order to use ECFCs as endothelial models for disease. Objectives: Therefore, we aimed to determine phenotypic and gene expression differences between control ECFCs. Methods: A total of 34 ECFC clones derived from 16 healthy controls were analyzed. The transcriptome of a selection of ECFC clones (n = 15) was analyzed by bulk RNA sequencing and gene set enrichment analysis. Gene expression was measured in all ECFC clones by quantitative polymerase chain reaction. Phenotypic profiling was performed and migration speed of the ECFCs was measured using confocal microscopy, followed by automated quantification of cell morphometrics and migration speed. Results: Through hierarchical clustering of RNA expression profiles, we could distinguish 2 major clusters within the ECFC cohort. Major differences were associated with proliferation and migration in cluster 1 and inflammation and endothelial-to-mesenchymal transition in cluster 2. Phenotypic profiling showed significantly more and smaller ECFCs in cluster 1, which contained more and longer Weibel–Palade bodies. Migration speed in cluster 1 was also significantly higher. Conclusion: We observed a range of different RNA expression patterns between ECFC clones, mostly associated with inflammation and clear differences in Weibel–Palade body count and structure. We developed a quantitative polymerase chain reaction panel that can be used for the characterization of ECFC clones, which is essential for the correct analysis of pathogenic mechanisms in vascular disorders.
AB - Background: Endothelial colony-forming cells (ECFCs) derived from patients can be used to investigate pathogenic mechanisms of vascular diseases like von Willebrand disease. Considerable phenotypic heterogeneity has been observed between ECFC clones derived from healthy donors. This heterogeneity needs to be well understood in order to use ECFCs as endothelial models for disease. Objectives: Therefore, we aimed to determine phenotypic and gene expression differences between control ECFCs. Methods: A total of 34 ECFC clones derived from 16 healthy controls were analyzed. The transcriptome of a selection of ECFC clones (n = 15) was analyzed by bulk RNA sequencing and gene set enrichment analysis. Gene expression was measured in all ECFC clones by quantitative polymerase chain reaction. Phenotypic profiling was performed and migration speed of the ECFCs was measured using confocal microscopy, followed by automated quantification of cell morphometrics and migration speed. Results: Through hierarchical clustering of RNA expression profiles, we could distinguish 2 major clusters within the ECFC cohort. Major differences were associated with proliferation and migration in cluster 1 and inflammation and endothelial-to-mesenchymal transition in cluster 2. Phenotypic profiling showed significantly more and smaller ECFCs in cluster 1, which contained more and longer Weibel–Palade bodies. Migration speed in cluster 1 was also significantly higher. Conclusion: We observed a range of different RNA expression patterns between ECFC clones, mostly associated with inflammation and clear differences in Weibel–Palade body count and structure. We developed a quantitative polymerase chain reaction panel that can be used for the characterization of ECFC clones, which is essential for the correct analysis of pathogenic mechanisms in vascular disorders.
UR - http://www.scopus.com/inward/record.url?scp=85191390238&partnerID=8YFLogxK
U2 - 10.1016/j.jtha.2024.03.018
DO - 10.1016/j.jtha.2024.03.018
M3 - Article
C2 - 38574861
AN - SCOPUS:85191390238
SN - 1538-7933
VL - 22
SP - 2027
EP - 2038
JO - Journal of Thrombosis and Haemostasis
JF - Journal of Thrombosis and Haemostasis
IS - 7
ER -