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Identification of Circulating Inflammatory Proteins Associated with Calcific Aortic Valve Stenosis by Multiplex Analysis

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Abstract

Calcific aortic valve stenosis (CAVS) is characterized by increasing inflammation and progressive calcification in the aortic valve leaflets and is a major cause of death in the aging population. This study aimed to identify the inflammatory proteins involved in CAVS and provide potential therapeutic targets. We investigated the observational and causal associations of 92 inflammatory proteins, which were measured using affinity-based proteomic assays. Firstly, the case–control cohort identified differential proteins associated with the occurrence and progression of CAVS. Subsequently, we delved into exploring the causal impacts of these associated proteins through Mendelian randomization. This involved utilizing genetic instruments derived from cis-protein quantitative loci identified in genome-wide association studies, encompassing a cohort of over 400,000 individuals. Finally, we investigated the gene transcription and protein expression levels of inflammatory proteins by single-cell and immunohistochemistry analysis. Multivariate logistic regression and spearman's correlation analysis showed that five proteins showed a significant positive correlation with disease severity. Mendelian randomization showed that elevated levels of two proteins, namely, matrix metallopeptidase-1 (MMP1) and sirtuin 2 (SIRT2), were associated with an increased risk of CAVS. Immunohistochemistry and single-cell transcriptomes showed that expression levels of MMP1 and SIRT2 at the tissue and cell levels were significantly higher in calcified valves than in non-calcified control valves. These findings indicate that MMP1 and SIRT2 are causally related to CAVS and open up the possibility for identifying novel therapeutic targets.

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Data Availability

The UK Biobank resource is open to legitimate researchers engaging in health-related studies for the public benefit. Researchers seeking access to this resource must register with UK Biobank by filling out the registration form available in the Access Management System (AMS-https://bbams.ndph.ox.ac.uk/ams/). The single-cell datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: https://www.ncbi.nlm.nih.gov/, PRJNA562645. For the current study, the datasets utilized and/or analyzed are obtainable from the corresponding author upon reasonable request.

Abbreviations

CAVS:

Calcific aortic valve stenosis

GWAS:

Genome-wide association studies

MR:

Mendelian randomization

SD:

Standard deviation

IL:

Interleukin

OR:

Odds ratio

SNP:

Single nucleotide polymorphism

IVW:

Inverse variance weighted

KEGG:

Kyoto encyclopedia of genes and genomes

GO:

Gene ontology

MMP1:

Matrix metallopeptidase-1

SIRT2:

Sirtuin 2

UMAP:

Uniform manifold approximation and projection

VDSCs:

Valve-derived stromal cells

VECs:

Valve endothelial cells

VICs:

Valve interstitial cells

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Acknowledgements

UKB is an open access resource. Bona fide researchers can apply to use the UKB dataset by registering and applying at http://ukbiobank.ac.uk/register-apply/.This research has been conducted using the UKB Resource under Application Number 68808 and we express our gratitude to the participants and those involved in building the resource

Funding

This study was supported by the National Key R&D Program of China (Grant No. 2021YFA0805100) and the National Natural Science Foundation of China (Grant Nos. 81861128025 and 81930014).

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Authors and Affiliations

  1. The Key Laboratory of Remodeling-Related Cardiovascular Diseases, Ministry of Education, Beijing Institute of Heart Lung and Blood Vessel Diseases, Beijing Anzhen Hospital, Capital Medical University, Anzhen Road, Chaoyang District, Beijing, 100029, China

    Rui Lin, Yuexin Zhu, Weiyao Chen, Zhiao Wang, Yuan Wang & Jie Du

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  1. Rui Lin
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  2. Yuexin Zhu
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Contributions

RL conceptualized the study, performed the experiments, carried out the analyses, and drafted the initial manuscript. YZ performed the Mendelian randomization analysis. WC and ZW obtained samples and contributed to data collection. JD and YW participated in the study design and reviewed and revised the manuscript.

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Correspondence to Yuan Wang or Jie Du.

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Use of human samples was approved by the Medical Ethical Committee of Beijing Anzhen Hospital and in concordance with the principles outlined in the Declaration of Helsinki. Informed consent was obtained.

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Handling Editor: Lu Cai.

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Lin, R., Zhu, Y., Chen, W. et al. Identification of Circulating Inflammatory Proteins Associated with Calcific Aortic Valve Stenosis by Multiplex Analysis. Cardiovasc Toxicol (2024). https://doi.org/10.1007/s12012-024-09854-5

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