During pregnancy the previously poor vasodilatory response of uterine artery endothelium (UA Endo) to agonists such as ATP is now enhanced through a greatly prolonged Capacitative Ca2+ entry (CCE), and this results in enhanced activation of eNOS. Since P-UA Endo's enhanced function now approaches that of other vessels, increases in uterine blood flow also occur. In preeclamptic (PE) subjects who often show elevated VEGF and TNF, this sustained Ca2+ response to ATP is also lost, along with a blunting of NO output. VEGF165 signals through ERK-1/2 to cause transient inhibitory phosphorylation of the Gap junction protein, Cx43 and loss of adapted CCE Ca2+ signaling. U0126 blockade can reverse this 0x43 phosphorylation and rescue pregnancy adapted Ca2+ burst function on subsequent challenge with ATP. Brief TNF mediated Src activation also directly phosphorylates CX43 on Tyr 265, causing Cx43 inhibition that is reversible by PP2. Long term.Src activation can also promote Tyr phosphorylation of ZO-1 and disassociation from Cx43, so causing Gap junction disassembly and 0x43 movement away from cell contact points and loss of pregnancy adapted CCE function. To that end:
Specific Aim 1 investigates the hypothesis that in P-UAEC, HUVEC and HUAEC that VEGF165 or TNF can mediate the dose dependent inhibition of sustained Ca2+ signaling through ERK or Src mediated direct CX43 protein phosphorylation.
Specific Aim 2 investigates the further hypothesis that these same VEGF or TNF mediated inhibitory effects in P-UAEC, HUVEC and HUAEC may also promote ZO-1 protein dissociation from CX43 wherever we see chronic activation of the Src pathway.
Specific Aim 3 translates Aims 1 and 2 back to intact vessels (UA Endo - pregnant sheep and HUV Endo or HUA Endo - normal human cords);by testing the hypothesis that the inhibitory actions of VEGF and TNF on endothelial Ca2+ observed in Aims 1 and 2 causes corresponding inhibition of NO production in vivo.
Specific Aim 4 tests the hypothesis that together even low doses of VEGF and TNF synergize to inhibit normal endothelial function in all vessels studied through the co-activation of Src and ERK-1/2 inhibitory pathways. A new therapy to treat PE may result from these studies.

Public Health Relevance

Preeclampsia is a disease of endothelial failure effecting 8-20% of the population, depending on ethnicity. We believe we have working model of the mechanisms underlying endothelial failure in preeclamptic pregnancy, and we propose to test that model. Our long term goal is that such studies could provide a proof of principle for targeting of new and novel therapies to treat this otherwise devastating disease.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Program Projects (P01)
Project #
5P01HD038843-12
Application #
8720795
Study Section
Special Emphasis Panel (ZHD1-DSR-Z)
Project Start
Project End
Budget Start
2014-06-01
Budget End
2015-05-31
Support Year
12
Fiscal Year
2014
Total Cost
$192,717
Indirect Cost
$62,469
Name
University of Wisconsin Madison
Department
Type
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715
Schreier, David A; Forouzan, Omid; Hacker, Timothy A et al. (2016) Increased Red Blood Cell Stiffness Increases Pulmonary Vascular Resistance and Pulmonary Arterial Pressure. J Biomech Eng 138:021012
Ampey, Bryan C; Morschauser, Timothy J; Ramadoss, Jayanth et al. (2016) Domain-Specific Partitioning of Uterine Artery Endothelial Connexin43 and Caveolin-1. Hypertension 68:982-8
Pastore, Mayra B; Talwar, Saira; Conley, Meghan R et al. (2016) Identification of Differential ER-Alpha Versus ER-Beta Mediated Activation of eNOS in Ovine Uterine Artery Endothelial Cells. Biol Reprod 94:139
Rozner, Ann E; Durning, Maureen; Kropp, Jenna et al. (2016) Macrophages modulate the growth and differentiation of rhesus monkey embryonic trophoblasts. Am J Reprod Immunol 76:364-375
Li, Yan; Wang, Kai; Zou, Qing-Yun et al. (2015) 2,3,7,8-Tetrachlorodibenzo-p-dioxin differentially suppresses angiogenic responses in human placental vein and artery endothelial cells. Toxicology 336:70-8
Morris, Erin A; Hale, Sarah A; Badger, Gary J et al. (2015) Pregnancy induces persistent changes in vascular compliance in primiparous women. Am J Obstet Gynecol 212:633.e1-6
Boeldt, Derek S; Grummer, Mary A; Yi, FuXian et al. (2015) Phosphorylation of Ser-279/282 and Tyr-265 positions on Cx43 as possible mediators of VEGF-165 inhibition of pregnancy-adapted Ca2+ burst function in ovine uterine artery endothelial cells. Mol Cell Endocrinol 412:73-84
Li, Yan; Wang, Kai; Zou, Qing-Yun et al. (2015) A possible role of aryl hydrocarbon receptor in spontaneous preterm birth. Med Hypotheses 84:494-7
Ampey, Bryan; Bird, Ian; Magness, Ron (2015) [307-POS]: Cx43 phosphorylation and the functionality of Cx43 gap junctions are moderated by cyclic nucleotide activity in UAECs and HUVECs. Pregnancy Hypertens 5:151-2
Anaya, Heather A; Yi, Fu-Xian; Boeldt, Derek S et al. (2015) Changes in Ca2+ Signaling and Nitric Oxide Output by Human Umbilical Vein Endothelium in Diabetic and Gestational Diabetic Pregnancies. Biol Reprod 93:60

Showing the most recent 10 out of 72 publications