A systems-approach reveals human nestin is an endothelial-enriched, angiogenesis-independent intermediate filament protein

Philip Dusart,Linn Fagerberg,Mathias Uhlén,Mete Civelek,Eike Struck,Ljubica Perisic,David-Alexandre Trégouët,Jacob Odeberg,Thomas Renné,Lynn M Butler,Ulf Hedin

doi:10.1038/s41598-018-32859-4

Abstract

The intermediate filament protein nestin is expressed during embryonic development, but considered largely restricted to areas of regeneration in the adult. Here, we perform a body-wide transcriptome and protein-profiling analysis to reveal that nestin is constitutively, and highly-selectively, expressed in adult human endothelial cells (EC), independent of proliferative status. Correspondingly, we demonstrate that it is not a marker for tumour EC in multiple malignancy types. Imaging of EC from different vascular beds reveals nestin subcellular distribution is shear-modulated. siRNA inhibition of nestin increases EC proliferation, and nestin expression is reduced in atherosclerotic plaque neovessels. eQTL analysis reveals an association between SNPs linked to cardiovascular disease and reduced aortic EC nestin mRNA expression. Our study challenges the dogma that nestin is a marker of proliferation, and provides insight into its regulation and function in EC. Furthermore, our systems-based approach can be applied to investigate body-wide expression profiles of any candidate protein.

Highlights

There are few studies of nestin expression in humans, with current knowledge almost entirely extrapolated from observations in animal models
This analysis indicated that nestin transcripts (NES) transcript expression is enriched in endothelial cells (EC) across organs, and we used protein profiling to confirm that nestin protein was expressed in the EC compartment of the human adult cerebral cortex, adrenal gland, thyroid, skeletal muscle, lung, tonsil, spleen, pancreas, salivary gland, esophagus, stomach, duodenum, colon, urinary bladder, adipose, placenta, ovary, testis and prostate (Fig. 1D)
Nestin was expressed in EC of the heart, kidney, and breast, in these tissues, consistent with previous reports[32,33,34], non-EC expression was observed in cardiomyocytes, renal glomeruli, and myoepithelial cells, respectively (Fig. 1E)

Summary

Introduction

There are few studies of nestin expression in humans, with current knowledge almost entirely extrapolated from observations in animal models. We perform a transcriptome and antibody-based profiling analysis of 37 human organs, which reveals body-wide EC-enriched nestin expression, independent of proliferative status. We analyse different tumour types to reveal that nestin is not a specific marker of tumour-associated EC, and its expression level is not an independent prognostic factor. We use primary human EC from four vascular beds to examine nestin spatial profiles under static and flow culture, and demonstrate an unexpected role for nestin in the inhibition of EC proliferation. We show reduced nestin expression in atherosclerotic plaque neovessels. Our study identifies nestin as a tissue-wide, EC-enriched IF, challenging its validity as a stem/progenitor or proliferative cell marker, its differential regulation between cell types and species, and its functional role in the adult vasculature

Methods

Results

Conclusion