Abstract
The role of primary cilia in mechanosensation is essential in endothelial cell (EC) shear responsiveness. Here, we find that venous, capillary, and progenitor ECs respond to shear stress invitro in a cilia-dependent manner. We then demonstrate thatprimary cilia assembly in human induced pluripotent stem cell (hiPSC)-derived ECs varies between different cell lines with marginal influence of differentiation protocol. hiPSC-derived ECs lacking cilia do not align to shear stress, lack stress fiber assembly, have uncoordinated migration during wound closure invitro, and have aberrant calcium influx upon shear exposure. Transcriptional analysis reveals variation in regulatory genes involved in ciliogenesis among different hiPSC-derived ECs. Moreover, inhibition of histone deacetylase 6 (HDAC6) activity in hiPSC-ECs lacking cilia rescues cilia formation and restores mechanical sensing. Taken together, these results show the importance of primary cilia in hiPSC-ECmechano-responsiveness and its modulation through HDAC6 activity varies among hiPSC-ECs.
Highlights
Endothelial cells (ECs) withstand a range of pressures and shear stress, mediating the exchange of oxygen, nutrients, and waste to surrounding tissue to support growth and tissue homeostasis
Quantification demonstrated that endothelial colony-forming cells (ECFCs), HUVECs, and HUAECs had similar aspect ratios (ARs) and nuclei orientation (Figure 1B)
Evaluating responses to 5 dynes/cm2, we found that while HUAECs and ECFCs oriented after 24 hr, HUVECs remained unaligned with a 1.2 ± 0.1 AR and 39 ± 1-degree orientation (Figure 1B)
Summary
Endothelial cells (ECs) withstand a range of pressures and shear stress, mediating the exchange of oxygen, nutrients, and waste to surrounding tissue to support growth and tissue homeostasis. Zebrafish studies have helped to identify primary cilia as a critical microtubule-based mechanosensory organelle (May-Simera and Kelley, 2012) that is indispensable for proper vascular development. An antenna-like structure composed of nine outer microtubule doublets, protrudes from the apical cell membrane and is present on ECs before the onset of flow. Cilia bend to changes in flow, resulting in the translation of physical forces to intracellular molecular signals (i.e., calcium influx), which acts to direct downstream vascular morphogenic events including angiogenesis (Goetz et al, 2014). Vascular tissue homeostasis relies on coordinated and deterministic cell migration, a process requiring dynamic cytoskeletal reorganization, and primary cilia presentation. Jones et al (2012) demonstrate that primary cilia defects can result in endothelial dysfunction including the ability to directionally migrate. While the importance of primary cilia has been shown in vivo and in vitro, the role of cilia presentation in the mechanosensation of developing ECs from human induced pluripotent stem cells (hiPSCs) has yet to be elucidated
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