The mechanosensory channel PIEZO1 functions upstream of angiopoietin/TIE/FOXO1 signaling in lymphatic development
Jing Du, … , Jing Jin, Susan E. Quaggin
Jing Du, … , Jing Jin, Susan E. Quaggin
Published May 15, 2024
Citation Information: J Clin Invest. 2024;134(10):e176577. https://doi.org/10.1172/JCI176577.
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The mechanosensory channel PIEZO1 functions upstream of angiopoietin/TIE/FOXO1 signaling in lymphatic development

Abstract

Lymphedema is a debilitating disease with no effective cure and affects an estimated 250 million individuals worldwide. Prior studies have identified mutations in piezo-type mechanosensitive ion channel component 1 (PIEZO1), angiopoietin 2 (ANGPT2), and tyrosine kinase with Ig-like and EGF-like domains 1 (TIE1) in patients with primary lymphedema. Here, we identified crosstalk between these molecules and showed that activation of the mechanosensory channel PIEZO1 in lymphatic endothelial cells (LECs) caused rapid exocytosis of the TIE ligand ANGPT2, ectodomain shedding of TIE1 by disintegrin and metalloproteinase domain–containing protein 17 (ADAM17), and increased TIE/PI3K/AKT signaling, followed by nuclear export of the transcription factor FOXO1. These data establish a functional network between lymphedema-associated genes and provide what we believe to be the first molecular mechanism bridging channel function with vascular signaling and intracellular events culminating in transcriptional regulation of genes expressed in LECs. Our study provides insights into the regulation of lymphatic function and molecular pathways involved in human disease.

Authors

Jing Du, Pan Liu, Yalu Zhou, Sol Misener, Isha Sharma, Phoebe Leeaw, Benjamin R. Thomson, Jing Jin, Susan E. Quaggin

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Figure 7

Yoda1 induces TIE1 shedding and ANGPT2 exocytosis via extracellular calcium influx.

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Yoda1 induces TIE1 shedding and ANGPT2 exocytosis via extracellular calc...
(A) Intracellular calcium levels in HDLECs treated with Yoda1 or vehicle were visualized using confocal microscopy with the cell-permeable Ca2+ indicator Fluo-8 AM. Scale bar: 50 μm. (B) Quantification of Yoda1-induced calcium influx in HDLECs pretreated with varying concentrations of the calcium chelator BAPTA. RFU, relative fluorescence units; T–T0, difference between the value measured at time point (T) and the value measured immediately prior to the treatment (T0). (C) Immunostaining for FOXO1, ANGPT2, and TIE1 in HDLECs treated with either vehicle or the calcium ionophore A23187 for 30 minutes. Arrows indicate the areas at cell-cell junctions where TIE1 shedding occurred. Scale bar: 50 μm. (D) Western blot analysis of TIE1 shedding and ANGPT2 exocytosis in HDLECs treated with vehicle or A23187. Each band represents a biological replicate sample (n = 3). (E) Western blot analysis of Yoda1-triggered TIE1 shedding and ANGPT2 exocytosis in the presence or absence of BAPTA. Each band represents a biological replicate sample (n = 3). Data are expressed as the mean ± SD. **P < 0.01 and ***P < 0.001, by 2-tailed, unpaired Student’s t test (D) and 2-way ANOVA followed by Tukey’s multiple-comparison test (E).