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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Research Article Development Vascular biology

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 2

TIE-mediated regulation of FOXO1 subcellular localization in vivo and in vitro.

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TIE-mediated regulation of FOXO1 subcellular localization in vivo and in...
(A) Whole mounts of E16.5 skin with lymphatic vessels stained for PROX1 and FOXO1. Nuclei within areas of LECs from WT control, Tie1WB–/–E13.5, and Tie2WB–/–E13.5 mice are indicated by arrowheads (branching points) and arrows (nonbranching points). Scale bars: 20 μm. (B) Quantification of FOXO1 localizations in PROX1+ LECs. Averaged values calculated from 3 mice in each group are presented. (C) P1 Tie1WB–/–E18.5 pups and their littermate controls were intraperitoneally injected with 1 μg/g BW Hepta-ANG1 or PBS. After a 30-minute period, pups were euthanized, and mesentery specimens were harvested and stained for PROX1 and FOXO1. Scale bars: 50 μm. (D) Quantification of FOXO1 localizations from 4 mice in each group. (E) FOXO1 staining of HDLECs transfected with siCtr, siTIE1, or siTIE2 for 48 hours and subsequently treated with Hepta-ANG1 (1 μg/mL) or PBS for 30 minutes. This experiment was replicated 3 times. Scale bar: 50 μm. (F) Quantification of FOXO1 localization in 3 fields of view selected from each group. (G) Western blot analysis of p-AKT levels in HDLECs transfected with siCtr, siTIE1, or siTIE2 and treated with either vehicle or Hepta-ANG1. 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 (F and G).