MicroRNA-425 and microRNA-155 cooperatively regulate atrial natriuretic peptide expression and cGMP production.

Sara Vandenwijngaert,Clara D Ledsky,Donald B Bloch,Obiajulu Agha,Connie Wu,Dongjian Hu,Ibrahim J Domian,Christopher Newton-Cheh,Aranya Bagchi,Emmanuel S Buys,Klaus Roemer

doi:10.1371/journal.pone.0196697

Abstract

AimsAtrial natriuretic peptide (ANP), secreted primarily by atrial cardiomyocytes, decreases blood pressure by raising cyclic 3’,5’-guanosine monophosphate (cGMP) levels and inducing vasorelaxation, natriuresis, and diuresis. Raising the level of ANP has been shown to be an effective treatment for hypertension. To advance the future development of an anti-microRNA (miR) approach to increasing expression of ANP, we investigated the regulation of NPPA expression by two miRs: miR-425 and miR-155. We examined whether miR-425 and miR-155 have an additive effect on the expression and function of ANP.Methods and resultsHuman embryonic stem cell-derived cardiomyocytes (hESC-CMs) were transfected with miR-425, miR-155, or a combination of the two miRs. Two days later, NPPA expression was measured using real time qPCR. Each of the miRs decreased NPPA expression over a wide range of concentrations, with a significant reduction at concentrations as low as 1 nM. The combination of miR-425 and miR-155 reduced NPPA expression to a greater extent than either miR-425 or miR-155 alone. An in vitro assay was developed to study the potential biological significance of the miR-induced decrease in NPPA expression. The cooperative effect of miR-425 and miR-155 on NPPA expression was associated with a significant decrease in cGMP levels.ConclusionsThese data demonstrate that miR-425 and miR-155 regulate NPPA expression in a cooperative manner. Targeting both miRNAs with anti-miRs (possibly at submaximal concentrations) might prove to be a more effective strategy to modulate ANP levels, and thus blood pressure, than targeting either miRNA alone.

Highlights

Hypertension is the leading modifiable risk factor for premature death and disability worldwide, and affects more than 1.3 billion individuals [1]
The objective of this study was to investigate whether miR-425 and miR-155 have an additive effect on the repression of NPPA gene expression
Human ESC-CMs were transfected with 1 nM, 5 nM, 10 nM, or 20 nM of miR-425 or miR-155, and NPPA mRNA levels were measured 48 hours later

Summary

Introduction

Hypertension is the leading modifiable risk factor for premature death and disability worldwide, and affects more than 1.3 billion individuals [1]. Genomic research has spurred tremendous progress in uncovering the genetics of blood pressure regulation and hypertension in humans, and has prompted the development of mechanism-based therapies. One of the pathways regulating blood pressure is the natriuretic peptide system. Natriuretic peptides mediate natriuresis, diuresis, and vasodilation by binding to the natriuretic peptide receptor 1 (NPR1), causing increased production of the second messenger cyclic guanosine 3’,5’-monophosphate (cGMP). The dual-acting drug LCZ696 (sacubitril-valsartan), which combines an inhibitor of neprilysin (an enzyme that degrades natriuretic peptides) with an inhibitor of the angiotensin receptor II, has been approved by the US Food and Drug Administration for the treatment of heart failure. In the PARAMETER and other clinical studies [4,5,6], LCZ696 was shown to lower blood pressure and pulse pressure, rekindling interest in the natriuretic peptide system as a therapeutic target for hypertension and cardiovascular disease

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Conclusion