Proteomic Approach Using DIA-MS Identifies Morphogenesis-Associated Proteins during Cardiac Differentiation of Human iPS Cells

Human-induced pluripotent stem cell (hiPSC)-derived cardiomyocytes
have potential applications in regenerative medicine. The quality
by design (QbD) approach enables the efficiency and quality assurance
in the manufacturing of hiPSC-derived products. It requires a molecular
understanding of hiPSC differentiation throughout the differentiation
process; however, information on cardiac differentiation remains limited.
Proteins associated with the early stages of cardiac differentiation
would be useful in the cardiomyocyte quality assessment. Here, we
performed quantitative proteomics of hiPSC intermediate cells in the
early phase of cardiac differentiation to better understand their
molecular characteristics. Proteomic profiles suggested that day 5–7
cells were in the morphogenetic stage of cardiac differentiation.
Trophoblast glycoprotein (TPBG) was the most up-regulated protein
in the morphogenetic stage; it was previously shown to be up-regulated
during differentiation into neural stem cells. Proteomics of TPBG-knockdown
cells revealed that TPBG is involved in cell proliferation and is
related to the cardiomyocyte yield, suggesting that it could be used
as a marker in QbD development. Our approach helps us understand the
molecular basis of hiPSC differentiation and could be a powerful tool
in QbD-based manufacturing.