Functionally Integrated Top-Down Proteomics for Standardized Assessment of Human Induced Pluripotent Stem Cell-Derived Engineered Cardiac Tissues

Three-dimensional (3D) human induced pluripotent stem cell-derived engineered cardiac tissues (hiPSC-ECTs) have emerged as a promising alternative to two-dimensional hiPSC-cardiomyocyte monolayer systems because hiPSC-ECTs are a closer representation of endogenous cardiac tissues and more faithfully...

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Bibliographic Details
Published in:Journal of proteome research 2021-02, Vol.20 (2), p.1424-1433
Main Authors: Melby, Jake A, de Lange, Willem J, Zhang, Jianhua, Roberts, David S, Mitchell, Stanford D, Tucholski, Trisha, Kim, Gina, Kyrvasilis, Andreas, McIlwain, Sean J, Kamp, Timothy J, Ralphe, J. Carter, Ge, Ying
Format: Article
Language:English
Subjects:
Cardiotoxicity
Cell Differentiation
Humans
Induced Pluripotent Stem Cells
Myocytes, Cardiac
Proteomics
Tissue Engineering
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Summary:Three-dimensional (3D) human induced pluripotent stem cell-derived engineered cardiac tissues (hiPSC-ECTs) have emerged as a promising alternative to two-dimensional hiPSC-cardiomyocyte monolayer systems because hiPSC-ECTs are a closer representation of endogenous cardiac tissues and more faithfully reflect the relevant cardiac pathophysiology. The ability to perform functional and molecular assessments using the same hiPSC-ECT construct would allow for more reliable correlation between observed functional performance and underlying molecular events, and thus is critically needed. Herein, for the first time, we have established an integrated method that permits sequential assessment of functional properties and top-down proteomics from the same single hiPSC-ECT construct. We quantitatively determined the differences in isometric twitch force and the sarcomeric proteoforms between two groups of hiPSC-ECTs that differed in the duration of time of 3D-ECT culture. Importantly, by using this integrated method we discovered a new and strong correlation between the measured contractile parameters and the phosphorylation levels of alpha-tropomyosin between the two groups of hiPSC-ECTs. The integration of functional assessments together with molecular characterization by top-down proteomics in the same hiPSC-ECT construct enables a holistic analysis of hiPSC-ECTs to accelerate their applications in disease modeling, cardiotoxicity, and drug discovery. Data are available via ProteomeXchange with identifier PXD022814.
ISSN:1535-3893
1535-3907
DOI:10.1021/acs.jproteome.0c00830