Human Peripheral Blood-Derived Endothelial Colony-Forming Cells Are Highly Similar to Mature Vascular Endothelial Cells yet Demonstrate a Transitional Transcriptomic Signature.

Anton G Kutikhin,Daria K Shishkova,Alexey E Tupikin,Elena A Velikanova,Marsel R Kabilov,Vera G Matveeva,Larisa V Antonova

doi:10.3390/cells9040876

Abstract

Endothelial colony-forming cells (ECFC) are currently considered as a promising cell population for the pre-endothelialization or pre-vascularization of tissue-engineered constructs, including small-diameter biodegradable vascular grafts. However, the extent of heterogeneity between ECFC and mature vascular endothelial cells (EC) is unclear. Here, we performed a transcriptome-wide study to compare gene expression profiles of ECFC, human coronary artery endothelial cells (HCAEC), and human umbilical vein endothelial cells (HUVEC). Characterization of the abovementioned cell populations was carried out by immunophenotyping, tube formation assay, and evaluation of proliferation capability while global gene expression profiling was conducted by means of RNA-seq. ECFC were similar to HUVEC in terms of immunophenotype (CD31+vWF+KDR+CD146+CD34-CD133-CD45-CD90-) and tube formation activity yet had expectedly higher proliferative potential. HCAEC and HUVEC were generally similar to ECFC with regards to their global gene expression profile; nevertheless, ECFC overexpressed specific markers of all endothelial lineages (NRP2, NOTCH4, LYVE1), in particular lymphatic EC (LYVE1), and had upregulated extracellular matrix and basement membrane genes (COL1A1, COL1A2, COL4A1, COL4A2). Proteomic profiling for endothelial lineage markers and angiogenic molecules generally confirmed RNA-seq results, indicating ECFC as an intermediate population between HCAEC and HUVEC. Therefore, gene expression profile and behavior of ECFC suggest their potential to be applied for a pre-endothelialization of bioartificial vascular grafts, whereas in terms of endothelial hierarchy they differ from HCAEC and HUVEC, having a transitional phenotype.

Highlights

Coronary artery bypass graft surgery remains an efficient and widespread surgical option to treat coronary artery disease [1], yet availability of autologous conduits such as internal mammary arteriesCells 2020, 9, 876; doi:10.3390/cells9040876 www.mdpi.com/journal/cellsCells 2020, 9, 876 or saphenous veins is often limited in patients with widespread atherosclerotic vascular disease or in those whose vessels are anatomically incompatible or have already been harvested for a previous procedure [2,3,4]
Recent studies report successful differentiation of peripheral blood mononuclear cells (PBMC) into endothelial colony-forming cells (ECFC) [12,13] which is efficient after percutaneous coronary intervention due to a mechanical injury provoking the release of ECFC precursors into the bloodstream [14]
We found that the baseline gene expression profile of ECFC is close to that of human coronary artery endothelial cells (HCAEC) and human umbilical vein endothelial cells (HUVEC) but expectedly different from subcutaneous adipose tissue-derived stromal vascular fraction (SAT-SVF), testifying to their utility for the seeding of tubular scaffolds before implantation to improve their short- and long-term performance

Summary

Introduction

Coronary artery bypass graft surgery remains an efficient and widespread surgical option to treat coronary artery disease [1], yet availability of autologous conduits such as internal mammary arteriesCells 2020, 9, 876; doi:10.3390/cells9040876 www.mdpi.com/journal/cellsCells 2020, 9, 876 or saphenous veins is often limited in patients with widespread atherosclerotic vascular disease or in those whose vessels are anatomically incompatible or have already been harvested for a previous procedure [2,3,4]. Allogeneic and xenogeneic blood vessels demonstrate only a limited efficiency due to the risk of transmissible disease, graft-versus-host disease, infection, and calcification [4,5] while prosthetic grafts fabricated from biostable synthetic polymers such as poly(ethylene terephthalate), expanded poly(tetrafluoroethylene), and polyurethanes show inferior patency rates in small-diameter applications due to poor endothelialization, low blood flow, and compliance mismatch, all resulting in intimal hyperplasia, thrombosis, or (pseudo)aneurysms [2,3,4,5] Both xenogeneic and biostable synthetic vascular conduits lack growth adaptation potential and demand repeated surgery and lead to unacceptable long-term outcomes [5,6]. ECFC exhibit high angiogenic [15,16]

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Conclusion