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. 2024 Mar 5;25(5):2999.
doi: 10.3390/ijms25052999.

Investigating the Role of Brain Natriuretic Peptide (BNP) and N-Terminal-proBNP in Thrombosis and Acute Ischemic Stroke Etiology

Rosanna Rossi  1   2   3 , Duaa Jabrah  1 , Andrew Douglas  1   2 , James Prendergast  1 , Abhay Pandit  2 , Michael Gilvarry  4 , Ray McCarthy  4 , Petra Redfors  5   6 , Annika Nordanstig  5   6 , Turgut Tatlisumak  5   6 , Erik Ceder  7 , Dennis Dunker  7 , Jeanette Carlqvist  7 , István Szikora  8 , Georgios Tsivgoulis  9 , Klearchos Psychogios  10 , John Thornton  11 , Alexandros Rentzos  7 , Katarina Jood  5   6 , Jesus Juega  12 , Karen M Doyle  1   2
Affiliations

Investigating the Role of Brain Natriuretic Peptide (BNP) and N-Terminal-proBNP in Thrombosis and Acute Ischemic Stroke Etiology

Rosanna Rossi et al. Int J Mol Sci. .

Abstract

The need for biomarkers for acute ischemic stroke (AIS) to understand the mechanisms implicated in pathological clot formation is critical. The levels of the brain natriuretic peptides known as brain natriuretic peptide (BNP) and NT-proBNP have been shown to be increased in patients suffering from heart failure and other heart conditions. We measured their expression in AIS clots of cardioembolic (CE) and large artery atherosclerosis (LAA) etiology, evaluating their location inside the clots, aiming to uncover their possible role in thrombosis. We analyzed 80 thrombi from 80 AIS patients in the RESTORE registry of AIS clots, 40 of which were of CE and 40 of LAA etiology. The localization of BNP and NT-BNP, quantified using immunohistochemistry and immunofluorescence, in AIS-associated white blood cell subtypes was also investigated. We found a statistically significant positive correlation between BNP and NT-proBNP expression levels (Spearman's rho = 0.668 p < 0.0001 *). We did not observe any statistically significant difference between LAA and CE clots in BNP expression (0.66 [0.13-3.54]% vs. 0.53 [0.14-3.07]%, p = 0.923) or in NT-proBNP expression (0.29 [0.11-0.58]% vs. 0.18 [0.05-0.51]%, p = 0.119), although there was a trend of higher NT-proBNP expression in the LAA clots. It was noticeable that BNP was distributed throughout the thrombus and especially within platelet-rich regions. However, NT-proBNP colocalized with neutrophils, macrophages, and T-lymphocytes, suggesting its association with the thrombo-inflammatory process.

Keywords: BNP; NT-proBNP; acute ischemic stroke; brain natriuretic peptide; stroke biomarkers; stroke etiology; thrombus.

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Conflict of interest statement

Authors M.G. and R.M. were employed by the company Cerenovus. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
(A) Violin plot showing no difference in BNP expression in clots from patients with LAA (N = 40) or CE stroke etiology (N = 40). (B) Violin plot showing no difference in NT-proBNP expression in clots from patients with LAA (N = 40) or CE stroke etiology (N = 40). Dashed lines represent the median, while dotted lines represent the interquartile ranges, Q1 (lower dotted lines) and Q3 (upper dotted lines). The p value from Mann–Whitney U-test analysis is indicated.
Figure 2
Figure 2
(A) MSB staining of the main clot components. (B,C) Expression of BNP and NT-proBNP in the same clot. BNP expression is located principally within platelet-rich regions, while NT-proBNP expression is only observed in nucleated cells. Higher magnification images are provided in the third row. In the fourth row, negative staining (A) and positive control staining images (pancreatic cancer tissue) for BNP (B) and proBNP (C) are shown. All images were captured using a 20× objective.
Figure 3
Figure 3
Colocalization analysis of NT-proBNP and WBC subtypes in AIS clots, showing individual signals (i–iii) and merged images (iv). Top row: T-lymphocytes (CD3); middle row: neutrophils (CD66b); bottom row: macrophages (CD68). All images were captured using a 60× objective (scale bar 5 μm).
See this image and copyright information in PMC

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