Microphysiological model of PIK3CA-driven vascular malformations reveals a role of dysregulated Rac1 and mTORC1/2 in lesion formation

Sci Adv. 2023 Feb 15;9(7):eade8939. doi: 10.1126/sciadv.ade8939. Epub 2023 Feb 15.

Abstract

Somatic activating mutations of PIK3CA are associated with development of vascular malformations (VMs). Here, we describe a microfluidic model of PIK3CA-driven VMs consisting of human umbilical vein endothelial cells expressing PIK3CA activating mutations embedded in three-dimensional hydrogels. We observed enlarged, irregular vessel phenotypes and the formation of cyst-like structures consistent with clinical signatures and not previously observed in cell culture models. Pathologic morphologies occurred concomitant with up-regulation of Rac1/p21-activated kinase (PAK), mitogen-activated protein kinase cascades (MEK/ERK), and mammalian target of rapamycin (mTORC1/2) signaling networks. We observed differential effects between alpelisib, a PIK3CA inhibitor, and rapamycin, an mTORC1 inhibitor, in mitigating matrix degradation and network topology. While both were effective in preventing vessel enlargement, rapamycin failed to reduce MEK/ERK and mTORC2 activity and resulted in hyperbranching, while inhibiting PAK, MEK1/2, and mTORC1/2 mitigates abnormal growth and vascular dilation. Collectively, these findings demonstrate an in vitro platform for VMs and establish a role of dysregulated Rac1/PAK and mTORC1/2 signaling in PIK3CA-driven VMs.

MeSH terms

  • Class I Phosphatidylinositol 3-Kinases / genetics
  • Class I Phosphatidylinositol 3-Kinases / metabolism
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Mechanistic Target of Rapamycin Complex 1 / metabolism
  • Mitogen-Activated Protein Kinase Kinases / metabolism
  • Sirolimus / pharmacology
  • TOR Serine-Threonine Kinases* / metabolism
  • Vascular Malformations* / metabolism
  • rac1 GTP-Binding Protein / metabolism

Substances

  • TOR Serine-Threonine Kinases
  • Mechanistic Target of Rapamycin Complex 1
  • Sirolimus
  • Mitogen-Activated Protein Kinase Kinases
  • Class I Phosphatidylinositol 3-Kinases
  • RAC1 protein, human
  • rac1 GTP-Binding Protein
  • PIK3CA protein, human