Abstract
Calcific aortic valve disease (CAVD) is the most common heart valve disease requiring intervention. Most research on CAVD has focused on inflammation, ossification, and cellular phenotype transformation. To gain a broader picture into the wide range of cellular and molecular mechanisms involved in this disease, we compared the total protein profiles between calcified and non-calcified areas from 5 human valves resected during surgery. The 1413 positively identified proteins were filtered down to 248 proteins present in both calcified and non-calcified segments of at least 3 of the 5 valves, which were then analyzed using Ingenuity Pathway Analysis. Concurrently, the top 40 differentially abundant proteins were grouped according to their biological functions and shown in interactive networks. Finally, the abundance of selected osteogenic proteins (osteopontin, osteonectin, osteocalcin, osteoprotegerin, and RANK) was quantified using ELISA and/or immunohistochemistry. The top pathways identified were complement system, acute phase response signaling, metabolism, LXR/RXR and FXR/RXR activation, actin cytoskeleton, mineral binding, nucleic acid interaction, structural extracellular matrix (ECM), and angiogenesis. There was a greater abundance of osteopontin, osteonectin, osteocalcin, osteoprotegerin, and RANK in the calcified regions than the non-calcified ones. The osteogenic proteins also formed key connections between the biological signaling pathways in the network model. In conclusion, this proteomic analysis demonstrated the involvement of multiple signaling pathways in CAVD. The interconnectedness of these pathways provides new insights for the treatment of this disease.
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Abbreviations
- APP:
-
Amyloid-β A4 protein
- a-SMA:
-
A-Smooth muscle actin
- FXR/RXR:
-
Farnesoid X receptor/retinoid X receptor
- LPC:
-
Lysophosphatidylcholine
- LXR/RXR:
-
Liver X receptor/retinoid X receptor
- MAC:
-
Membrane attack complex
- MYC:
-
Myc proto-oncogene protein
- OSTCN*:
-
Osteocalcin
- OSTP*:
-
Osteopontin
- TNR11*:
-
RANK/CD265
- SNP:
-
Single nucleotide polymorphisms
- SPARC*:
-
Osteonectin
- TGFB1:
-
Transformation growth factor-β1
- TR11B*:
-
Osteoprotegerin
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Acknowledgements
The authors thank Katie Brown for blinded IHC scoring. This study was supported in part by NIH grants T32 HL07812 and R21 HL104377.
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Dr. Reardon serves as a consultant for Medtronic, Boston Scientific, Abbott Medical, and Gore Medical; all fees for such are to his department and there is no overlap between the consulting work and the research presented here.
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Research use of the surgically-resected tissues was approved by the Institutional Review Boards at Houston Methodist Hospital and Rice University and was in accordance with the ethical standards of the IRBs and with the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.
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Informed consent was obtained and details that might disclose the identity of patients were removed from samples and the study.
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In Abbreviations: *Whenever possible, the UniProt (UniProt KB) protein name is used for consistency.
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Han, R.I., Hu, C.W., Loose, D.S. et al. Differential proteome profile, biological pathways, and network relationships of osteogenic proteins in calcified human aortic valves. Heart Vessels 37, 347–358 (2022). https://doi.org/10.1007/s00380-021-01975-z
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DOI: https://doi.org/10.1007/s00380-021-01975-z