SUMMARY
Preeclampsia (PE) is a common, life-threatening complication in pregnancy, characterized by high blood
pressure putting the mother at risk of eclampsia and kidney dysfunction. PE affects about 1–5% of
pregnancies and a major cause of maternal and fetal deaths. The hallmarks of PE are the decrease in
trophoblast invasion and abnormal remodeling of the spiral arteries; as well as an
angiogenic/antiangiogenic imbalance. The abnormal placentation will, most likely, cause reduced feto-
maternal blood flow and increased oxidative stress resulting in compromised function. There is no clear
pathogenesis or cure, thus an effective treatment for PE is an unmet medical need. The primary goal of
this proposal is to develop novel cationic organofluorine hydrazones for mitochondria-targeted
antioxidant therapy in preeclampsia. Three key structural features will be combined to effective
antioxidants: (i) a hydrazone core for improved antioxidant potential, (ii) fluorine incorporation for
increased lipophilicity and membrane permeability, and (iii) introduction of quaternary ammonium salts
(QASs) for the delivery of the lipophilic cation through the mitochondrial membrane. Based on our
preliminary data and the results of in silico evaluations, we will design and synthesize QAS-containing
organofluorine antioxidants that will be subjected to primary screening for their in vitro antioxidant activity
in biochemical assays. The compounds that will show significant radical scavenging properties will be
evaluated in cell-based and in vivo assays. Human primary trophoblast cells (normal or PE pregnancy)
will be tested whether augmenting cell-redox function chemically by the proposed antioxidants will
reduce (i) cell injury, (ii) mitochondrial stress, (iii) HIF1a production, and (iv) downstream anti-angiogenic
response to hypoxia-reoxygenation. The same assays will also be carried out in human endothelial cells
exposed to hypoxia-reoxygenation. In addition, time-pregnant mice will be subjected to hypoxia, to
induce the main features of PE, (hypertension, proteinuria and oxidative stress), and treated with the
synthetic antioxidants and the mice will be evaluated for blood pressure, renal function, histologic
damage, 3-nitrotyrosine (3-NT) tissue immunoreactivity, inflammation markers and plasma biomarkers.
Finally, mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy will be used to
map the presence of the new cationic hydrazones in cells and tissues. These studies may lead to the
development of new effective antioxidant compounds, which antagonize ROS/RNS that prevent the
activation of the HIF1¿ pathway and therefore improve angiogenic balance and reduce the systemic
effects of PE. Regarding the educational/training aspects, this work will provide a multi-environmental
and interdisciplinary training to undergraduate students who will participate in each step of the project
and be exposed to the research atmosphere at two different institutions.