The pathophysiological role and therapeutic relevance of vitamin D in preeclampsia

• Applicants: Professor Dr. Ralf Dechend; Professorin Dr. Frauke von Versen-Höynck
• Subject Area: Gynaecology and Obstetrics ; Reproductive Medicine, Urology
• Term: from 2014 to 2020
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 263521300

• Final Report Year: 2021

Final Report Abstract

Within this DFG-funded project we were able to gain significant further insights into the effects of VD on the capacity, integrity and functionality of different cell types, especially ECFC, HUVEC and trophoblast cells. In our first study we report that HUVEC from preeclamptic pregnancies showed reduced migration and tubule formation. Placental villous explant conditioned medium derived from 2% O2 incubations reduced HUVEC migration. Unexpectedly, VD did not improve HUVEC function or VEGF gene expression. Because of these results we wanted to compare the functionality and ability with a special subtype of endothelial progenitor cells which are a promising tool for vascular repair. Here, we report ECFC form cellcell contacts with each other and that VD increases endothelial integrity and barrier function under inflammatory conditions. While ECFC barrier is disrupted under inflammatory conditions this effect was rescued by vitamin D treatment, leading to higher stability of an ECFC monolayer. Further, VD enhanced ECFC mobilization towards directional migration. In addition, immunocytochemistry, qRT-PCR and immunoblotting analysis showed that VD increased endothelial interconnections through VE-cadherin junctions and by impacting cell dynamics through cofilin and VE cadherin phosphorylation. Our results suggest that vitamin D treatment efficiently counteracts inflammation resulting in higher ECFC barrier integrity. This provides evidence of a new beneficial effect of VD for ECFC homeostasis. Because of these results we further investigated the ability of ECFC to invade into established monolayers and capillary tubule-like structures of HUVEC in the presence/absence of fetal cord serum from uncomplicated or PE pregnancies, and tested the ability of VD to modulate the serum-mediated effects. PE cord serum reduced the invasion of fetal ECFCs into HUVEC monolayers or tubule networks. VD attenuated these effects of PE fetal serum on endothelial functional properties. Immunocytochemical studies revealed involvement of VE-cadherin contacts in interactions between ECFC and mature fetal endothelial cells. These data appear consistent with lines of evidence that vitamin D has anti-PE effects. However, how VD impacts these pathways remains unclear and the underlying mechanisms need to be further elucidated in future studies. Based on this we additionally analyzed the expression of pro- (VEGF, PLGF) and antiangiogenic factors (sFLT1, sENG) in preeclamptic ECFC and a trophoblast cell line under hypoxic conditions, but in neither of these groups strong differences could be detected in comparison to controls. The role of VD in vivo was investigated in three different established rodent models for PE. The transgenic PE model is induced by local and circulating upregulation of the RAS, the RUPP model by ischemia of the uteroplacental unit and the AT1-AA infusion model by auto-immune dysregulation. In all three models VD was an important modulator, however the effect of VD depletion in the transgenic model was not present. Otherwise we observed a profound effect on blood pressure and anti-angiogenesis and less pronounced effect on fetal intrauterine growth retardation. In a prospective population-based pregnancy study, low VD levels were associated with an increased risk of PE. Low VD levels were associated with disturbed function of regulatory T cells and a state of anti-angiogenesis.

Publications

• Vitamin D3 depletion aggravates hypertension and target-organ damage. J Am Heart Assoc 2015 Jan 28;4(2). pii: e001417
  Andersen LB, Przybyl L, Haase N, von Versen-Höynck F, Qadri F, Jørgensen JS, Sorensen GL, Fruekilde P, Poglitsch M, Szijarto I, Gollasch M, Peters J, Muller DN, Christesen HT, Dechend R
  
• Vitamin D supplementation improves pathophysiology in a rat model of preeclampsia. Am J Physiol Regul Integr Comp Physiol. 2016 Feb 15;310(4):R346-54
  Faulkner JL, Cornelius DC, Amaral LM, Harmon AC, Cunningham MW Jr, Darby MM, Ibrahim T, Thomas DS, Herse F, Wallukat G, Dechend R, LaMarca B
  
• Vitamin D3 depletion does not affect key aspects of the preeclamptic phenotype in a transgenic rodent model for preeclampsia. J Am Soc Hypertens. 2016 Jul;10(7):597-607.e1
  Andersen LB, Golic M, Przybyl L, Sorensen GL, Jørgensen JS, Fruekilde P, von Versen-Höynck F, Herse F, Højskov CS, Christesen HT, Haase N, Dechend R
  
• Impaired Functional Capacity of Fetal Endothelial Cells in Preeclampsia. PLoS One 2017 May 23;12(5):e0178340
  Brodowski L, Burlakov J, Hass S, von Kaisenberg C, von Versen-Höynck F
  
• Vitamin D supplementation reduces some AT1-AA- induced downstream targets implicated in preeclampsia including hypertension. Am J Physiol Regul Integr Comp Physiol. 2017 Jan 1;312(1):R125-R131
  Faulkner JL, Amaral LM, Cornelius DC, Cunningham MW, Ibrahim T, Heep A, Campbell N, Usry N, Wallace K, Herse F, Dechend R, LaMarca B
  
• The role of vitamin D3 in cell-cell interaction of fetal endothelial progenitor cells and umbilical cord endothelial cells in a preeclampsia-like model. Am J Physiol- Cell Physiol. 2019 Aug 1;317(2):C348- C357
  Brodowski L, Schröder-Heurich B, von Hardenberg S., Vu T, von Kaisenberg CS, von Versen-Höynck F
  
• Vitamin D3 improves endothelial barrier integrity and counteracts inflammatory effects on endothelial progenitor cells. FASEB J. 2019 Aug;33(8):9142- 9153
  Schröder-Heurich B, von Hardenberg S, Brodowski L, Kipke B, Meyer N, Borns K, von Kaisenberg CS, Brinkmann H, Claus P, von Versen-Höynck F