Importance of smooth muscle p110alpha/AKT/FOXO1 signaling for the development of abdominal aortic aneurysm
Authors: Vantler M , Schorscher M , Berghausen E M , Moore Iv J B , Wong D , Li Z , Gnatzy-Feik L , Zierden M , Mehrkens D , Adam M , Sengle G , Boor P , Maegdefessel L , Baldus S , Rosenkranz S 
Published: November 2023

Abstract

Background
Abdominal aortic aneurysms (AAA) are characterized by loss of vascular smooth muscle cells (SMCs) and extracellular matrix (ECM) degradation. The catalytic PI 3-kinase isoform p110α signals downstream of growth factor receptors. Mice harboring an SMC-specific p110α deficiency (SM-p110α-/-) are characterized by impaired SMC proliferation and survival.

Aim
We hypothesized that p110α deficiency in SMCs exacerbates the development and progression of AAA as it promotes SMC loss and thus weakens the aortic wall in healthy and diseased animals. Therefore, we investigated how p110α deficiency in SMCs affects AAA formation, vascular integrity as well as downstream signaling in SMCs.

Methods and results
Medial thickness of the abdominal aorta in SM-p110α-/- mice was significantly decreased compared to wild-type (WT) littermate controls (29.0±3.1µm vs. 42.5±4.1µm). Morphological characterization of the aortic wall by transmission electron microscopy revealed impaired elastic fibers, detached SMCs and apoptotic cells in SM-p110α-/- mice. SMC differentiation markers including calponin and myosin heavy chain were downregulated in the aortic wall of SM-p110α-/- mice. Western blots demonstrated that p110α deficiency impaired the expression of elastin. Consequently, elastin fibers were characterized by reduced fiber width and length. In addition, growth factor stimulated proliferation of p110α-/- SMCs was significantly impaired compared to WT controls. AAA development in SM-p110α-/- mice and WT littermates were studied using the porcine pancreatic elastase (PPE) model. PPE was infused into the infrarenal aorta. Ultrasound examination of the aorta revealed an increased aortic diameter in all PPE-treated mice. However, the increase in aortic diameter was significantly elevated (p&lt;0.05) in SM-p110α-/- mice (70.2±10.8%, n=9) compared to WT animals (42.4±6.0%, n=10). Furthermore, PPE treated SM-p110-/- mice were characterized by increased ECM degradation, decreased vascular remodelling, and less proliferating cells. Mechanistically, growth factor-dependent phosphorylation and inactivation of crucial phenotypic modulators of SMCs - FOXO-1, -3 and -4 - were shown to be significantly decreased in p110α-/- SMCs compared to WT cells. Specific inhibition of FOXO1 rescued the impaired proliferation of p110α-/- SMCs and enhanced the expression of elastin and calponin. Analysis of FOXO expression in humans revealed that FOXO1 was significantly upregulated in AAA patients compared to healthy individuals.

Conclusion
p110α deficiency in SMCs promotes the development and progression of AAA, as p110α controls crucial SMC functions which are important for structural processes in the aortic wall. Thereby, SMC proliferation is controlled by p110α dependent phosphorylation and inactivation of FOXO1. Since FOXO1 is upregulated in AAA patients, the p110α/AKT/FOXO1 signaling cascade may provide a key target for modulating aortic wall stability in AAA.