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The effect of cell elongation and alignment via topographical micropatterning on the endothelial transcriptome

ABSTRACT: The aim of this study was to investigate the effect of endothelial cell (EC) elongation and alignment, independent of fluid shear stress (FSS), on the endothelial transcriptome. Endothelial cells are continuously exposed to a variety of biomechanical stimuli and must adapt their structure, gene expression, and phenotype to maintain vascular homeostasis. FSS-induced changes in EC morphology are highly associated with functional phenotype. However, to fully investigate EC mechanotransduction pathways, it is essential to decouple cell morphology from FSS. We, therefore, investigated the effects of elongated and aligned endothelial monolayers on the endothelial transcriptome. Baboon carotid artery endothelial cells (ECs) were seeded on either fully non-patterned, planar polyurethane culture substrates or micropatterned (3 µm wide anisotropic ridges and grooves, 1 µm depth) polyurethane culture substrates. We found EC elongation and alignment within static culture induced a highly differential gene transcriptome with a multitude of pro-inflammatory genes downregulated by ECs on micropatterned versus planar substrates. Collectively, our data support that endothelial morphology independently holds a critical role in the genetic transcriptome and may contribute to cardiovascular diseases and inflammation.

ORGANISM(S): Papio anubis

PROVIDER: GSE247648 | GEO | 2024/11/30

REPOSITORIES: GEO

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Project description:Tissue Factor Pathway Inhibitor-2 is Induced by Fluid Shear Stress in Vascular Smooth Muscle Cells and Affects Cell Proliferation and Survival Introduction: Vascular smooth muscle cells (SMCs) are exposed to fluid shear stress (FSS) after interventional procedures such as balloon-angioplasty. Whereas the effects of hemodynamic forces on endothelial cells are explored in detail, the influence of FSS on smooth muscle cell function is poorly characterized. Here, we investigated the effect of FSS on SMC gene expression and function. Methods: Laminar FSS of arterial level (14 dynes/cm2) was applied to SMC cultures for 24 h in a parallel-plate flow chamber. The effect of FSS on gene expression was first screened with microarray technology and the results further verified by real time (RT) PCR and immunoblotting. Protein expression was also studied in the rat carotid artery after balloon-injury and DNA synthesis and apoptosis was examined in SMCs in vitro. Results: Microarrays identified tissue-pathway inhibitor-2 (TFPI-2) as the most differentially expressed gene by FSS in cultured SMCs. The regulatory effect of FSS on the expression of TFPI-2 was confirmed by RT-PCR and immunobloting demonstrating a more than 400-fold increase in TFPI-2 expression in SMCs exposed to FSS compared to static controls and a consistent upregulation at the protein level. Functionally, SMC proliferation was decreased by FSS and recombinant TFPI-2 was found to inhibit SMC proliferation and induce SMC apoptosis as indicated by activation of caspase-3. In vivo, TFPI-2 expression was found to be up-regulated 5, 10 and 20 h after rat carotid balloon-injury and immunohistochemistry demonstrated TFPI-2 protein in luminal SMCs exposed to FSS in rat carotid intimal hyperplasia 10 days after balloon-injury. Conclusion: FSS strongly influence gene expression in cultured SMCs and induce TFPI-2 expression, which is also expressed after rat carotid balloon injury in luminal SMCs exposed to FSS. Functionally, TFPI-2 may play an important role in vessel wall repair by regulating SMC proliferation and survival. Further studies are needed to elucidate the mechanisms by which TFPI-2 control SMC function. 3 x 2 samples from a paired fluid shear stress experiment on smooth muscle cells.

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The effect of cell elongation and alignment via topographical micropatterning on the endothelial transcriptome

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