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Spatial Multiplexed Protein Profiling of Cardiac Ischemia-Reperfusion Injury
Luyan Yao1, Funan He1, Quanyi Zhao1,2
1State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing (L.Y., F.H., Q.Z., D.L., S.F., M.Z., X.Z., B.Z., L.W.).
Insights
Understanding cardiac cell spatial organization is key to minimizing heart attack reperfusion injury. Targeting H3K9me3 in endothelial cells shows promise for reducing cardiac damage and remodeling after myocardial infarction.
Area of Science:
- Cardiovascular Biology
- Cellular and Molecular Medicine
- Proteomics and Genomics
Background:
- Reperfusion therapy is vital for salvaging heart muscle after myocardial infarction but can cause ischemia-reperfusion injury (IRI).
- Limited knowledge of cardiac cell spatial organization impedes the development of interventions to mitigate IRI effects.
Purpose of the Study:
- To characterize the spatial distribution and dynamics of cardiac cell phenotypes and communities following IRI using imaging mass cytometry.
- To identify potential therapeutic targets for minimizing IRI-induced cardiac damage.
Main Methods:
- Utilized imaging mass cytometry on mouse left ventricle sections across 12 cardiac segments and 8 time points.
- Analyzed 251 multiplexed images, identifying over 197,000 single cells grouped into 23 distinct cell communities.
- Investigated protein posttranslational modifications, focusing on H3K9me3 in endothelial cells.
Main Results:
- Revealed heterogeneous and dynamic cardiac cellular architecture following IRI.
- Identified increased H3K9me3 in endothelial cells as a key regulatory response during the middle stage of IRI.
- Demonstrated that manipulating H3K9me3 levels (silencing Suv39h1 or overexpressing Kdm4d) attenuated cardiac dysfunction and remodeling post-IRI.
Conclusions:
- Mapped the spatiotemporal heterogeneity of cardiac cellular phenotypes after IRI.
- Uncovered H3K9me3 in endothelial cells as a potential therapeutic target to alleviate pathological remodeling in myocardial IRI.
Background:
Reperfusion therapy is critical to myocardial salvage in the event of a myocardial infarction but is complicated by ischemia-reperfusion injury (IRI). Limited understanding of the spatial organization of cardiac cells, which governs cellular interaction and function, has hindered the search for targeted interventions minimizing the deleterious effects of IRI.
Methods:
We used imaging mass cytometry to characterize the spatial distribution and dynamics of cell phenotypes and communities in the mouse left ventricle following IRI. Heart sections were collected from 12 cardiac segments (basal, mid-cavity, apical, and apex of the anterior, lateral, and inferior wall) and 8 time points (before ischemia [I-0H], and postreperfusion [R-0H, R-2H, R-6H, R-12H, R-1D, R-3D, R-7D]), and stained with 29 metal-isotope-tagged antibodies. Cell community analysis was performed on reconstructed images, and the most disease-relevant cell type and target protein were selected for intervention of IRI.
Results:
We obtained a total of 251 multiplexed images, and identified 197 063 single cells, which were grouped into 23 distinct cell communities based on the structure of cellular neighborhoods. The cellular architecture was heterogeneous throughout the ventricular wall and exhibited swift changes following IRI. Analysis of proteins with posttranslational modifications in single cells unveiled 13 posttranslational modification intensity clusters and highlighted increased H3K9me3 (tri-methylated lysine 9 of histone H3) as a key regulatory response in endothelial cells during the middle stage of IRI. Erasing H3K9 methylation, by silencing its methyltransferase Suv39h1 or overexpressing its demethylase Kdm4d in isolated endothelial cells, attenuated cardiac dysfunction and pathological remodeling following IRI. in vitro, H3K9me3 binding significantly increased at endothelial cell function-related genes upon hypoxia, suppressing tube formation, which was rescued by inhibiting H3K9me3.
Conclusions:
We mapped the spatiotemporal heterogeneity of cellular phenotypes in the adult heart upon IRI, and uncovered H3K9me3 in endothelial cells as a potential therapeutic target for alleviating pathological remodeling of the heart following myocardial IRI.
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