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Urinary extracellular vesicle mRNA analysis of sodium chloride cotransporter in hypertensive patients under different conditions

Abstract

Urinary extracellular vesicles (UEV) mainly derive from cells of the urogenital tract and their cargo (proteins, nucleic acids, lipids, etc.) reflects their cells of origin. Na chloride cotransporter (NCC) is expressed at the kidney level in the distal convoluted tubule, is involved in salt reabsorption, and is the target of the diuretic thiazides. NCC protein has been recognized and quantified in UEV in previous studies; however, UEV NCC mRNA has never been studied. This study aimed to identify and analyze NCC mRNA levels in primary aldosteronism (PA). The rationale for this investigation stems from previous observations regarding NCC (protein) as a possible biomarker for the diagnosis of PA. To evaluate modulations in the expression of NCC, we analyzed NCC mRNA levels in UEV in PA and essential hypertensive (EH) patients under different conditions, that is, before and after saline infusion, anti-aldosterone pharmacological treatment, and adrenal surgery. NCC mRNA was measured by RT-qPCR in all the samples and was regulated by volume expansion. Its response to mineralocorticoid receptor antagonist was correlated with renin, and it was increased in PA patients after adrenalectomy. NCC mRNA is evaluable in UEV and it can provide insights into the pathophysiology of distal convolute tubule in different clinical conditions including PA.

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Fig. 1: NCC mRNA measured in different conditions.
Fig. 2: NCC mRNA level in MRA treatment.
Fig. 3: NCC mRNA level after adrenalectomy.

Data availability

The datasets generated for this study will not be made publicly available due to privacy policy. Requests to access the datasets should be directed to FP, francesca.pizzolo@univr.it.

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Acknowledgements

We would like to acknowledge the special contributions of our beloved friend and colleague, Sarath Kiran Channavajjhala, who left us too early. His intelligence, dedication, and enthusiasm will always shine in our hearts. This study was performed (in part) at the Laboratorio Universitario di Ricerca Medica (LURM) Research Center at the University of Verona. TEM imaging was carried out at the Department of Neurosciences, Biomedicine and Movement Sciences at the University of Verona with the assistance of Dr. Paolo Bernardi.

Funding

This research was funded by the Italian Ministry for University and Research, project “Ricerca di Base” (to FP). The funder had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

Authors

Contributions

OO, AC, and FP contributed to conception and design of the study. LB, FM, NT, and DD performed the experiments. LB, OO, FP, AC, PB, SF and FM interpreted the results. NT, AC and LB prepared the figures. LB, FP, SF and AC drafted the manuscript. LB, DD, GLS, GS, FM analysed data. All authors contributed to manuscript revision, read, and approved the final version for submission.

Corresponding author

Correspondence to Francesca Pizzolo.

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This study was carried out in accordance with recommendations of the Ethics Committee of our Institution (Azienda Ospedaliera Universitaria Integrata, Verona, Italy), who approved the protocol. Written and informed consent were obtained from patients in accordance with the Declaration of Helsinki.

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The authors declare no competing interests.

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Pizzolo, F., Bertolone, L., Castagna, A. et al. Urinary extracellular vesicle mRNA analysis of sodium chloride cotransporter in hypertensive patients under different conditions. J Hum Hypertens (2022). https://doi.org/10.1038/s41371-022-00744-3

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