Dataset Information

Proteomics of novel iPSC-derived vascular endothelial cells reveal extensive similarity with an immortalized human endothelial cell line

ABSTRACT: The vascular endothelium constitutes the inner lining of the blood vessel and is directly involved in the control of blood fluidity, vascular tone, platelet aggregation, regulation of immunity, inflammation, and angiogenesis. Malfunction and injuries of the endothelium can cause cardiovascular diseases, which are the leading causes of death globally. The impairment of the endothelium is a hallmark in other diseases as well such as sepsis, stroke, tumor growth, insulin resistance, venous thrombosis, and chronic kidney failure. Generation of effective sources to replace injured endothelial cells (ECs) could have significant clinical impact and somatic cell sources like peripheral or cord blood cannot credibly supply enough endothelial cell progenitors for multitude of treatments. Pluripotent stem cells are a promising source for a reliable EC supply that have the potential to repair, reconstruct, replace damaged cells, and revascularize ischemic areas in order to restore tissue function and treat vascular diseases. We have developed methods to differentiate induced pluripotent stem cells (iPSCs) efficiently and robustly across multiple iPSC lines into non-tissue-specific pan vascular ECs (iECs) with high purity. These iECs present with canonical endothelial cell markers and exhibit measures of endothelial cell functionality with uptake of acetylated low-density lipoprotein (Dil-Ac-LDL) and tube formation. Using proteomic analysis, we revealed the iECs are more proteomically similar to established umbilical vein ECs (HUVECs) than to iPSCs. Post-translational modifications (PTMs) were most shared between HUVECs and iECs, and potential targets for increasing the proteomic similarity of iECs to HUVECs were identified. PTM analysis can be used in the future to glean additional insights and generate novel hypotheses. Here we demonstrate an efficient robust method to differentiate iPSCs into functional ECs, and for the first time provide a comprehensive protein expression profile of iECs, which indicates their similarities with a widely used immortalized HUVECs, allowing for further mechanistic studies of EC development, signaling and metabolism for future regenerative applications.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Stem Cell

SUBMITTER: Sarah Parker

LAB HEAD: Sarah Parker

PROVIDER: PXD038493 | Pride | 2023-07-05

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	1D26_DIA_SantosSareen_03n14_1.raw	Raw
	1D26_DIA_SantosSareen_03n14_2.raw	Raw
	1D26_DIA_SantosSareen_03n14_3.raw	Raw
	1D26_DIA_SantosSareen_E362_EDi028-A_1.raw	Raw
	1D26_DIA_SantosSareen_E362_EDi028-A_2.raw	Raw

Items per page:

1 - 5 of 29

Publications

Proteomics of novel induced pluripotent stem cell-derived vascular endothelial cells reveal extensive similarity with an immortalized human endothelial cell line.

Ariyasinghe Nethika R NR de Souza Santos Roberta R Gross Andrew A Aghamaleky-Sarvestany Arwin A Kreimer Simion S Escopete Sean S Parker Sarah J SJ Sareen Dhruv D

Physiological genomics 20230612 8

The vascular endothelium constitutes the inner lining of the blood vessel, and malfunction and injuries of the endothelium can cause cardiovascular diseases as well as other diseases including stroke, tumor growth, and chronic kidney failure. Generation of effective sources to replace injured endothelial cells (ECs) could have significant clinical impact, and somatic cell sources like peripheral or cord blood cannot credibly supply enough endothelial cell progenitors for multitude of treatments. ...[more]

PMID: 37306406

Similar Datasets

Project description:Atherosclerotic plaques tend to form in the major arteries at certain predictable locations. As these arteries vary in atherosusceptibility, inter-arterial differences in endothelial cell (EC) biology are of considerable interest. To explore the origin of differences observed between typical atheroprone and atheroresistant arteries, we used DNA microarrays to compare gene expression profiles of harvested porcine coronary (CECs) and iliac (IECs) artery ECs grown in static culture out to passage four. Fewer differences were observed between the transcriptional profiles of CECs and IECs in culture compared to in vivo, suggesting that most differences observed in vivo were due to distinct environmental cues in the two arteries. One-class significance of microarrays revealed that most in vivo interarterial differences disappeared in culture, as fold differences after passaging were not significant for 85% of genes identified as differentially expressed in vivo at a 5% false discovery rate. However, the three homeobox genes HOXA9, HOXA10, and HOXD3 remained under-expressed in coronary endothelium for all passages by at least 9, 8, and 2-fold, respectively. Continued differential expression despite removal from the in vivo environment suggests that primarily heritable or epigenetic mechanism(s) influence transcription of these three genes. Quantitative real-time polymerase chain reaction confirmed expression ratios for seven genes associated with atherogenesis and over- or under-expressed by 3-fold in CECs relative to IECs. The present study provides evidence that both local environment and vascular bed origin modulate gene expression in arterial endothelium. The transcriptional differences observed here may provide new insights into pathways responsible for coronary artery susceptibility. Endothelial cells were freshly harvested from right coronary and iliac arteries from four pigs. Cells were cultured out to passage four. RNA was isolated after each passage and expression profiles were obtained using oligo microarrays.