While endothelial dysfunction is known to be a part of preeclampsia (PE), the molecular basis for endothelial dysfunction remains unclear. Current therapies for the associated hypertension are still largely targeted to relaxation of vascular smooth muscle and yet this is not sufficient to fully control the disorder. New strategies are needed based on novel approaches to therapeutic intervention. This will only be possible given a clearly identified new therapeutic target and a means to screen it at the level of endothelial function. Such large scale screening approaches require large numbers of high purity, well characterized cells from PE subjects. Second round testing in resistance level vessels can then be undertaken on a realistic scale before moving towards drug refinement and trials in human subjects. The problem remains that human uterine artery tissue from normal and PE pregnancy are not sufficiently available to undertake the initial screen, and smaller vessel biopsy samples are insufficient to yield cells of high enough purity or quantity for drug panel screening. Our preliminary data suggests that while vascular endothelial function is indeed maximized in the uterine artery in normal pregnancy, these same mechanisms are also observed and maximized in other vascular beds including fetoplacental vessels such as the umbilical vein. Further, these signaling events and associated NO production in the endothelium of normal cords also appear to have failed in PE subjects. Thus, in order to establish if it is possible to use HUVEC from PE subjects as suitable cells in such a drug screen we propose:
Sp Aim 1 : Hypothesis: The observed sustained [Ca2+]i bursts in responses to ATP in intact UV Endo from normal pregnancy are due to TRPC3 and CX43 activation, and losses of Ca2+ burst associated with strategies aimed at TRPC3 or CX43 inhibition replicate the lower/blunted Ca2+ and NO responses seen in PE subjects.
Sp Aim 2 (Transitional to Aim 3): Hypothesis: PE associated dysfunction in PE derived UV Endo or PE derived HUVEC (passage 3) compared to that from normal subjects is not due to simple changes in the relative levels of expression of P2Y2 receptors, G proteins, PLC, IP3 receptors, TRPC channels, or connexins.
Sp Aim 3 : Hypothesis: In short term culture, the sustained [Ca2+]i burst responses to ATP in HUVEC from normal pregnancy due to CX43 potentiation of IP3 sensitive TRPC3 activation are retained, and are lacking in cells derived from PE subjects. Further, inhibition of either IP3 mediated TRPC3 activation or CX43 mediated cell-cell communication in HUVEC from normal subjects replicates the loss of function seen in HUVEC from PE subjects, but does not further inhibit the function of HUVEC from PE subjects. Given success, the ready availability of cords and large yields of UV Endo cells from PE subjects means that future large scale drug testing could be performed, even with additional focus on targeted/disadvantaged subpopulations of the public. This could allow more selective intervention with new drugs or new applications of existing drugs that impact on this signaling pathway, and so relieve the devastating effects of preeclampsia.
Preeclampsia (PE) is a potentially devastating disease of pregnancy that can threaten the life of both mother and the unborn child. It affects as many as 20% of the population. Even mild PE is dangerous and can cause preterm (early) birth and this then gives the baby an increased risk to develop several diseases such as blood pressure, diabetes and obesity as an adult. We have shown in mothers with PE that the cells lining the umbilical cord blood vessels have reduced function and so may be a readily available and useful tool to screen dugs for therapy. The aim of this project is to show our suspicions are correct. If we are right we may be able to come up with a drug-screening plan to develop future treatment. Having more complete knowledge of how cell function is regulated in umbilical vein from both normal and PE pregnancy will certainly take us one step closer to overcoming this potentially devastating disease that effects so many in this country and beyond.
|Boeldt, Derek S; Hankes, Amanda C; Alvarez, Roxanne E et al. (2014) Pregnancy programming and preeclampsia: identifying a human endothelial model to study pregnancy-adapted endothelial function and endothelial adaptive failure in preeclamptic subjects. Adv Exp Med Biol 814:27-47|
|Boeldt, Derek S; Grummer, Mary A; Magness, Ronald R et al. (2014) Altered VEGF-stimulated Ca2+ signaling in part underlies pregnancy-adapted eNOS activity in UAEC. J Endocrinol 223:1-11|
|Krupp, Jennifer; Boeldt, Derek S; Yi, Fu-Xian et al. (2013) The loss of sustained Ca(2+) signaling underlies suppressed endothelial nitric oxide production in preeclamptic pregnancies: implications for new therapy. Am J Physiol Heart Circ Physiol 305:H969-79|
|Bird, I M; Boeldt, D S; Krupp, J et al. (2013) Pregnancy, programming and preeclampsia: gap junctions at the nexus of pregnancy-induced adaptation of endothelial function and endothelial adaptive failure in PE. Curr Vasc Pharmacol 11:712-29|