#1610 FGF23-mediated hypertension via augmented calcium entry in vascular smooth muscle cells
Authors: Muñoz-Castañeda Juan R , Garcia-Saez Raquel , Lopez-Baltanas Rodrigo , Romero Ana Torralbo , Ortiz Maria Encarnacion Rodriguez , Ureña Juan , Fernandez-Tenorio Miguel , Jurado Daniel , Leiva Fernando , de Mier M Victoria Pendon-Ruiz , Rodriguez Mariano , Rodelo-Haad Cristian 
Published: May 2024

Background and Objectives

Ongoing studies are shedding light on the potential connection between FGF23 and hypertension. In experimental settings, the administration of recombinant FGF23 in mice has been demonstrated to induce hypertension by elevating serum sodium, along with an increase in NCC receptors in the kidney. Clinical evidences further supports pro-hypertensive effects of high circulating levels of FGF23. Our group has previously demonstrated a direct influence of FGF23 on increased arterial stiffness by inducing a phenotypic transition in vascular smooth muscle cells (VSMCs) from a contractile to a synthetic phenotype. This study aimed comprehensively to evaluate, both in vivo and in vitro, whether FGF23 directly leads to hypertension through the stimulation of intracellular calcium entry in vascular cells.

Methods

To ascertain whether FGF23 contributes to hypertension through a direct impact on blood vessels, recombinant FGF23 (15 µg/day) was administered for 28 days via Alzet pumps. Blood pressure was meticulously measured in these animals, and we investigated alterations in vascular remodeling and calcium channels, as well as proteins associated with vascular contraction such as Orai1, STIM1, PKD1, SERCA2a, and AGTR1. In vitro experiments included the evaluation of the effect of elevated levels of recombinant FGF23 (2 ng/ml) over 9 days on intracellular calcium entry stimulated by Ang2 (100 nM) or Thapsigargin (2.5 µM). We used the FURA method to conduct these experiments. To elucidate the in vitro pro-hypertensive mechanisms of FGF23, studies were conducted with the administration of inhibitors targeting FGF Receptors 1-3 (AZD4547, 150 nM), FGF Receptor-4 (BLU9931, 10 nM), and Erk1/2 phosphorylation (PD98059, 10 μM). Additionally, changes in Orai1, STIM1, PKD1, SERCA2a, and AGTR1 were also evaluated in these VSMCs.

Results

After 28 days of recombinant FGF23 treatment, no significant differences were observed in serum sodium levels, yet there was a marked increase in urinary phosphorus excretion. Systolic and diastolic blood pressure values showed a progressive significant elevation in animals exposed to high levels of FGF23. The expression of AGTR1 and PKD1 levels significantly increased after FGF23 administration for 28 days (Figure). However, Orai1 and STIM1 in the thoracic aorta exhibited no significant differences following FGF23 treatment.

In VSMCs, the exposure to recombinant FGF23 for 9 days resulted in increased calcium entry upon Ang2 stimulation (Figure). This effect was markedly diminished after inhibiting FGFR1-3 receptors and Erk1/2 phosphorylation (Figure). Notably, the inhibition of FGF23 receptor 4 (FGFR4) had no impact on the calcium entry induced by FGF23 (Figure). While Orai1, STIM1, IP3R, and TRPV5 in VSMCs remained unaffected by recombinant FGF23 administration, PKD1, SERCA2, and AGTR1 levels were significantly elevated compared to control cells (Figure). The addition of Thapsigargin in a Ca2+-free culture medium to VSMCs incubated with recombinant FGF23 for 9 days led to increased Ca2+ efflux from endoplasmic reticulum.

Conclusion

Elevated levels of FGF23 directly enhanced calcium entry in VSMCs. This effect is mediated by FGFR1-3 receptors, Erk1/2 phosphorylation, and the upregulation of related protein to cell contraction such as SERCA2a, PKD1 or AGTR1. These modifications could be responsible of FGF23-associated hypertension in the experimental model.