Preeclampsia is the leading cause of mortality and morbidity in pregnancy, and, still, it is a condition without a cure. Aberrant placental development and growth due to inappropriate trophoblast and/or endothelial cell (EC) function, are the underlying causes. FK506-binding protein like (FKBPL) is emerging as an important mechanism in the pathogenesis of preeclampsia1 and a predictive and diagnostic biomarker2. In this study, we aimed to determine whether the FKBPL-based therapeutic mimetic peptide, AD-01, can abrogate impairment in trophoblast proliferation, oxidative stress and EC migration in vitro, in 2D and 3D microfluidic models of preeclampsia.
First trimester trophoblast cells, ACH-3Ps, were exposed to various preeclamptic stimuli in 2D, including hypoxia (dimethyloxalylglycine, DMOG, 1 mM), inflammation (tumour necrosis factor, TNF-α, 10 ng/ml) or mitochondrial dysfunction (Rho-6G, 1 μg/ml). AD-01 (100 mM), was added as a treatment at the same time, for 48 h. Trophoblast proliferation and uric acid concentrations were measured using MTT and Uric Acid Assay Kit, respectively. Human microvascular endothelial cells (HMEC-1) were treated ± FKBPL siRNA or exposed to inflammatory or hypoxic conditions using macrophage-condition medium (MCM) or DMOG (1mM), respectively, ± AD-01 (100mM). A 3D microfluidics chip was developed. EC migration and 3D cellular protein expression was determined.
AD-01 rescued the impaired ACH-3Ps proliferation induced by DMOG (p<0.05) or Rho-6G (p<0.001). AD-01 also abrogated the TNF-α-mediated uric acid increase (p<0.001). FKBPL protein expression was reduced following DMOG treatment (p<0.05), whereas AD-01 normalised both HIF-1α (p<0.01) and FKBPL expression (p<0.001). MCM increased both HIF-1α (p<0.05) and FKBPL (p<0.01) expression, whereas AD-01 abrogated this effect (p<0.05-0.001). Suppression of FKBPL with siRNA or via MCM/DMOG, stimulated HMEC-1 migration within the chip (p<0.05-0.001).
FKBPL-based therapeutic peptide, AD-01, restored the angiogenic imbalance via FKBPL, and could be a viable treatment option for preeclampsia, with dual utility capable of rescuing trophoblast and EC dysfunction.