Reference glycan structure libraries of primary human cardiomyocytes and pluripotent stem cell-derived cardiomyocytes reveal cell-type and culture stage-specific glycan phenotypes

• Published in: Journal of molecular and cellular cardiology Vol. 139; pp. 33 - 46
• Main Authors: Ashwood, Christopher, Waas, Matthew, Weerasekera, Ranjuna, Gundry, Rebekah L.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.02.2020
• Subjects: Cardiomyocytes; Cells, Cultured; Female; Glycan structures; Glycomics; Glycosylation; Heart; hPSC-CM; Humans; Induced Pluripotent Stem Cells - metabolism; Male; Mass spectrometry; Myocytes, Cardiac - metabolism; Phenotype; Polysaccharides - chemistry; Polysaccharides - metabolism; Principal Component Analysis; Protein glycosylation; Time Factors; Tissue Fixation; Heart; hPSC-CM; hiPSC-CM; CFG; Neu5Ac; MALDI; MS; CDG; MeCN; Cardiomyocytes; LacDiNAc; Protein glycosylation; Neu5Gc; PGC; PVDF; PCA; WGA; LC; ESI; GlcNAc; Glycan structures; Mass spectrometry; ConA
• ISSN: 0022-2828; 1095-8584
• DOI: 10.1016/j.yjmcc.2019.12.012

Cell surface glycoproteins play critical roles in maintaining cardiac structure and function in health and disease and the glycan-moiety attached to the protein is critical for proper protein folding, stability and signaling [1]. However, despite mounting evidence that glycan structures are key modulators of heart function and must be considered when developing cardiac biomarkers, we currently do not have a comprehensive view of the glycans present in the normal human heart. In the current study, we used porous graphitized carbon liquid chromatography interfaced with mass spectrometry (PGC-LC-MS) to generate glycan structure libraries for primary human heart tissue homogenate, cardiomyocytes (CM) enriched from human heart tissue, and human induced pluripotent stem cell derived CM (hiPSC-CM). Altogether, we established the first reference structure libraries of the cardiac glycome containing 265 N- and O-glycans. Comparing the N-glycome of CM enriched from primary heart tissue to that of heart tissue homogenate, the same pool of N-glycan structures was detected in each sample type but the relative signal of 21 structures significantly differed between samples, with the high mannose class increased in enriched CM. Moreover, by comparing primary CM to hiPSC-CM collected during 20–100 days of differentiation, dynamic changes in the glycan profile throughout in vitro differentiation were observed and differences between primary and hiPSC-CM were revealed. Namely, >30% of the N-glycome significantly changed across these time-points of differentiation and only 23% of the N-glycan structures were shared between hiPSC-CM and primary CM. These observations are an important complement to current genomic, transcriptomic, and proteomic profiling and reveal new considerations for the use and interpretation of hiPSC-CM models for studies of human development, disease, and drug testing. Finally, these data are expected to support future regenerative medicine efforts by informing targets for evaluating the immunogenic potential of hiPSC-CM and harnessing differences between immature, proliferative hiPSC-CM and adult primary CM. [Display omitted] •First reference structure libraries of the human cardiac glycome•Significant glycosylation differences among cell types within the heart•Cardiomyocytes preferentially exhibit less mature glycans compared to whole heart tissue•Glycosylation is dynamic throughout in vitro differentiation•Differences between glycans in primary cardiomyocytes and hiPSC-CM are revealed