Microphysiological vascular malformation model reveals a role of dysregulated Rac1 and mTORC1/2 in lesion formation.

Aw, W. Y.; Cho, C.; Wang, H.; Cooper, A. H.; Doherty, E. L.; Rocco, D.; Huang, S. A.; Kubik, S.; Whitworth, C. P.; Armstrong, R.; Hickey, A. J.; Griffith, B.; Kutys, M. L.; Blatt, J.; Polacheck, W. J.

doi:10.1101/2022.09.03.506415

bioRxiv · 10.1101/2022.09.03.506415

Microphysiological vascular malformation model reveals a role of dysregulated Rac1 and mTORC1/2 in lesion formation.

Aw, W. Y.·Cho, C.·Wang, H.·Cooper, A. H.·Doherty, E. L.·Rocco, D.·Huang, S. A.·Kubik, S.·Whitworth, C. P.·Armstrong, R.·Hickey, A. J.·Griffith, B.·Kutys, M. L.·Blatt, J.·Polacheck, W. J.

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Abstract

Somatic activating mutations of PIK3CA are associated with the development of vascular malformations (VMs). Here, we describe a microfluidic model of PIK3CA-driven VMs consisting of human umbilical vein endothelial cells (HUVECs) expressing PIK3CA activating mutations embedded in 3D hydrogels. We observed enlarged and irregular vessel phenotypes, consistent with clinical signatures and concomitant with PI3K-driven upregulation of Rac1/PAK, MEK/ERK, and mTORC1/2 signaling. We observed differential effects between Alpelisib, a PIK3CA inhibitor, and Rapamycin, an mTORC1 inhibitor, in mitigating matrix degradation and vascular network topology. While both drugs are effective in preventing vessel enlargement, Alpelisib suppressed mTORC2-dependent AKT1 phosphorylation and MEK/ERK signaling. Rapamycin failed to reduce MEK/ERK and mTORC2 activity and resulted in vascular hyperbranching, while inhibiting PAK, MEK1/2, and mTORC1/2 signaling mitigates abnormal growth and vascular dilation. Collectively, these findings establish an in vitro platform for modeling VMs and confirm a role of dysregulated Rac1/PAK and mTORC1/2 signaling in PIK3CA-driven VMs.

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Aw, W. Y., Cho, C., Wang, H., Cooper, A. H., Doherty, E. L., Rocco, D., Huang, S. A., Kubik, S., Whitworth, C. P., Armstrong, R., Hickey, A. J., Griffith, B., Kutys, M. L., Blatt, J., Polacheck, W. J.. 2022-09-04. Microphysiological vascular malformation model reveals a role of dysregulated Rac1 and mTORC1/2 in lesion formation.. https://doi.org/10.1101/2022.09.03.506415

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