Multiomics Method Enabled by Sequential Metabolomics and Proteomics for Human Pluripotent Stem-Cell-Derived...

Area of Science:

• Cardiovascular Biology
• Stem Cell Research
• Proteomics and Metabolomics

Background:

• Human pluripotent stem-cell-derived cardiomyocytes (hPSC-CMs) hold potential for disease modeling and regenerative therapies.
• Current hPSC-CM cultures do not fully replicate adult cardiomyocyte properties in vivo.
• A deeper understanding of hPSC-CM molecular biology is needed.

Purpose of the Study:

• To establish a multiomics method for simultaneous analysis of metabolome and proteome from hPSC-CMs.
• To create multidimensional molecular profiles of hPSC-CMs.
• To provide a foundation for advancing hPSC-CM biology research.

Main Methods:

• Developed a sequential extraction technique for metabolites and proteins from hPSC-CM monolayer cultures.
• Utilized high-resolution mass spectrometry for analysis.
• Integrated proteomic and metabolomic data to generate network profiles.

Main Results:

• Successfully annotated 205 metabolites/lipids and 4319 proteins from 10^6 cells with high reproducibility.
• Identified 310 pathways, with 40 significantly overrepresented (FDR-corrected p ≤ 0.05).
• Key pathways identified include protein synthesis, ATP generation (oxidative phosphorylation), and muscle contraction.

Conclusions:

• The developed multiomics method offers deep coverage of metabolites and proteins in hPSC-CMs.
• This approach provides a multidimensional view of hPSC-CM phenotype.
• The method serves as a technological foundation to enhance understanding and application of hPSC-CMs.