BACKGROUND:
In developmental and pathological tissues, nascent vessel networks generated by angiogenesis require further pruning/regression to delete nonfunctional endothelial cells (ECs) by apoptosis and migration. Mechanisms underlying EC apoptosis during vessel pruning remain elusive. TMEM215 (transmembrane protein 215) is an endoplasmic reticulum–located, 2-pass transmembrane protein. We have previously demonstrated that TMEM215 knockdown in ECs leads to cell death, but its physiological function and mechanism are unclear.
METHODS:
We characterized the role and mechanism of TMEM215 in EC apoptosis using HUVECs by identifying its interacting proteins with immunoprecipitation-mass spectrometry. The physiological function of TMEM215 in ECs was assessed by establishing a conditional knockout mouse strain. The role of TMEM215 in pathological angiogenesis was evaluated by tumor and choroidal neovascularization models. We also tried to evaluate its translational value by delivering a
Tmem215
siRNA using nanoparticles in vivo.
RESULTS:
TMEM215
knockdown in ECs induced apoptotic cell death. We identified the chaperone BiP as a binding partner of TMEM215, and TMEM215 forms a complex with and facilitates the interaction of BiP with the BH (BCL-2 homology) 3-only proapoptotic protein BIK (BCL-2 interacting killer).
TMEM215
knockdown triggered apoptosis in a BIK-dependent way and was abrogated by BCL-2. Notably,
TMEM215
knockdown increased the number and diminished the distance of mitochondria-associated endoplasmic reticulum membranes and increased mitochondrial calcium influx. Inhibiting mitochondrial calcium influx by blocking the IP
3
R (inositol 1,4,5-trisphosphate receptor) or MCU (mitochondrial calcium uniporter) abrogated
TMEM215
knockdown–induced apoptosis.
TMEM215
expression in ECs was induced by physiological laminar shear stress via
EZH2
downregulation. In EC-specific
Tmem215
knockout mice, induced
Tmem215
depletion impaired the regression of retinal vasculature characterized by reduced vessel density, increased empty basement membrane sleeves, and increased EC apoptosis. Moreover, EC-specific
Tmem215
ablation inhibited tumor growth with disrupted vasculature. However,
Tmem215
ablation in adult mice attenuated lung metastasis, consistent with reduced
Vcam1
expression. Administration of nanoparticles carrying
Tmem215
siRNA also inhibited tumor growth and choroidal neovascularization injury.
CONCLUSIONS:
TMEM215
, which is induced by blood flow-derived shear stress via downregulating
EZH2
, protects ECs from BIK-triggered mitochondrial apoptosis mediated by calcium influx through mitochondria-associated ER membranes during vessel pruning, thus providing a novel target for antiangiogenic therapy.