Wortmannin targeting phosphatidylinositol 3‐kinase suppresses angiogenic factors in shear‐stressed endothelial cells

Abstract

Modifications on shear stress‐based mechanical forces are associated with pathophysiological susceptibility and their effect on endothelial cells (EC) needs to be better addressed looking for comprehending the cellular and molecular mechanisms. This prompted us to better evaluate the effects of shear stress in human primary venous EC obtained from the umbilical cord, using an in vitro model to mimic the laminar blood flow, reaching an intensity 1–4 Pa. First, our data shows there is a significant up‐expression of phosphatidylinositol 3‐kinase (PI3K) in shear‐stressed cells culminating downstream with an up‐phosphorylation of AKT and up‐expression of MAPK‐ERK, concomitant to a dynamic cytoskeleton rearrangement upon integrin subunits (α4 and ß 3) requirements. Importantly, the results show there is significant involvement of nitric oxide synthase (eNOS), nNOS, and vascular endothelial growth factors receptor 2 (VEGFR2) in shear‐stressed EC, while cell cycle‐related events seem to being changed. Additionally, although diminution of 5‐hydroxymethylcytosine in shear‐stressed EC, suggesting a global repression of genes transcription, the promoters of PI3K and eNOS genes were significantly hydroxymethylated corroborating with their respective transcriptional profiles. Finally, to better address, the pivotal role of PI3K in shear‐stressed EC we have revisited these biological issues by wortmannin targeting PI3K signaling and the data shows a dependency of PI3K signaling in controlling the expression of VGFR1, VGFR2, VEGF, and eNOS, once these genes were significantly suppressed in the presence of the inhibitor, as well as transcripts from Ki67 and CDK2 genes. Finally, our data still shows a coupling between PI3K and the epigenetic landscape of shear‐stressed cells, once wortmannin promotes a significant suppression of ten‐11 translocation 1 (TET1), TET2, and TET3 genes, evidencing that PI3K signaling is a necessary upstream pathway to modulate TET‐related genes. In this study we determined the major mechanotransduction pathway by which blood flow driven shear stress activates PI3K which plays a pivotal role on guaranteeing endothelial cell phenotype and vascular homeostasis, opening novel perspectives to understand the molecular basis of pathophysiological disorders related with the vascular system.

PI3K/AKT signaling drives titanium-induced angiogenic stimulus

“…2 and 3). Finally, to test the hypothesis that PI3K/AKT drives angiogenic factors in response to the Ti-enriched medium, we subjected the cells to a chemical inhibition of PI3K using well-known Wortmannin, as it is widely proposed by others [43], and further investigated the behavior of VEGF (Fig. 4a), VEGFR1 (Fig.…”

“…Thus, we reasonably suggest that PI3K/AKT signaling participates in the angiogenic phenotype of ECs responding to the Ti-enriched medium. We previously reported that the major mechanotransduction pathway activates PI3K, which plays a pivotal role in guaranteeing EC phenotype and vascular homeostasis [43], and Ti-release displays a very similar effect, mainly in response to Wo/DAE.…”

Identifying potential drug targets and candidate drugs for COVID-19: biological networks and structural modeling approaches

“…In the present study, we repurposed Wortmannin as a candidate drug that can inhibit PI3K signaling and receptor tyrosine kinase (ERBB4) mediated macropinocytosis, to control virus-cell proliferation and may enhance ERBB4 functions namely alveolarization, morphogenesis, and AEC2 differentiation. Wortmannin is a covalent inhibitor of phosphatidylinositol-3-kinase (PI3K) and related enzymes include the mammalian target of rapamycin (mTOR), DNA-dependent protein kinase catalytic subunit (DNA-PKcs), and mitogen-activated protein kinase (MAPK) 94,95 . In humans, PI3K activates cell signaling pathways in the head and neck, urinary tract, cervical, ovarian, and lung cancer.…”

The Exploitation of Liposomes in the Inhibition of Autophagy to Defeat Drug Resistance

“…3-methyladenine (3-MA) is one of the first used inhibitors of autophagy (Xie et al, 2016), under starving conditions it suppresses autophagy by inhibiting phosphatidylinositol 3kinase, catalytic subunit type 3 (PI3KC3), on the other hand, with nutrients it favors autophagy by using PI3KC1 inhibition (Laha et al, 2019). Because 3-MA was shown to be effective only at high doses, other derivatives were investigated, as Wortmannin, a fungal metabolite that binds to the catalytic site of PI3Ks, and SF1126 (Gomes et al, 2019;Qin et al, 2019). SF1126 is a novel and potent dual inhibitor of phosphatidylinositol 3-kinase and bromodomain-containing protein 4 oncogenes.…”

“…Here we will review literature acquisitions concerning the use of liposomes to improve the bioavailability of autophagy inhibitors. Once at the target the drug can be release passively or upon a trigger that can originate from the microenvironment, for example the acidic environment of tumor tissues, or by the outside (light, temperature, magnetic field, ultrasounds) (Needham et al, 2000;Alhmady et al, 2012;Chen et al, 2013;Adriyanov et al, 2014;Luo et al, 2016a;Luo et al, 2016b). Passive release does not require further liposome functionalization, whereas in the case of triggered release liposome have to be engineered ad hoc.…”