Expandable Arterial Endothelial Precursors from Human CD34+ Cells Differ in Their Proclivity to Undergo an Endothelial-to-Mesenchymal Transition

Abstract

SummaryArterial diseases continue to pose a major health concern but in vitro studies are limited because explanted cells can exhibit poor proliferative capacity and a loss of specificity. Here, we find that two transcription factors, MYCN and SOX17, induce and indefinitely expand in culture precursors of human arterial endothelial cells (expandable arterial endothelial precursors [eAEPs]). The eAEPs are derived from CD34+ cells found in umbilical cord blood or adult bone marrow. Independent eAEP lines differ in their proclivity to undergo an endothelial-to-mesenchymal transition (EndoMT), a hallmark event in a broad array of vascular diseases and disorders. Some cell lines spontaneously become mesenchymal over time in culture, an effect exacerbated by inhibition of the fibroblast growth factor receptor, while others do not readily convert. These distinctions were exploited to identify genes that correlate with resistance to an EndoMT and to elucidate transcriptional changes that underpin the transition.

The molecular basis for functional divergence of duplicated SOX factors controlling endoderm formation and left-right patterning in zebrafish

“…Cloning for in vitro production of mRNAs pCS2+hSOX17 was cloned from PB-TRE3G-SOX17 (Miller et al, 2018) using XhoI and XbaI restriction sites. PB-TRE3G-SOX17 was a gift from David Vereide (Addgene plasmid # 104541 ; http://n2t.net/addgene:104541 ; RRID:Addgene_104541) Sox17 and sox32 deletion and domain switch constructs (Table S1) were generated using pCS2+sox17 and pCS2+myc-sox32 by PCR amplification using Q5 polymerase (NEB) using described primers (Table S2) followed by re-circularisation or insertion using T4 ligase (Invitrogen).…”

“…pCS2+ hSOX17 was cloned from PB-TRE3G-SOX17 (Miller et al, 2018) using XhoI and XbaI restriction sites. PB-TRE3G-SOX17 was a gift from David Vereide (Addgene plasmid # 104541 ; http://n2t.net/addgene:104541 ; RRID:Addgene_104541)…”

Single cell and lineage tracing studies reveal the impact of CD34+ cells on myocardial fibrosis during heart failure

“…Over the past decade, it is well known that resident cells that express c-Kit, Sca-1 and CD34 cannot differentiate into cardiomyocytes [ 9 – 12 ], but they do have an ability to differentiate into endothelial cells and other non-cardiomyocytes in response to myocardial injury [ 13 ]. For instance, CD34 + cells exist in tissues, circulating blood and bone marrow, which may serve as a source of different types of cells in pathological status [ 14 ]. Bone marrow-derived CD34 + cell therapy has received great attention for its potential applications in cardiovascular disease [ 15 – 17 ].…”