TY - JOUR
T1 - Reference glycan structure libraries of primary human cardiomyocytes and pluripotent stem cell-derived cardiomyocytes reveal cell-type and culture stage-specific glycan phenotypes
AU - Ashwood, Christopher
AU - Waas, Matthew
AU - Weerasekera, Ranjuna
AU - Gundry, Rebekah L.
N1 - Funding Information:
This work was supported by the National Institutes of Health [ R01-HL126785 and R01-HL134010 to RLG; F31-HL140914 to MW]; Funding sources were not involved in study design, data collection, interpretation, analysis or publication. Mass spectrometry analyses were performed using instrumentation in the Center for Biomedical Mass Spectrometry Research at the Medical College of Wisconsin. Tissue sectioning was performed by the Children's Research Institute Histology core lab. Flow cytometry analyses were performed using instrumentation in the Blood Center of Wisconsin Flow Cytometry Core.
Funding Information:
This work was supported by the National Institutes of Health [R01-HL126785 and R01-HL134010 to RLG; F31-HL140914 to MW]; Funding sources were not involved in study design, data collection, interpretation, analysis or publication. Mass spectrometry analyses were performed using instrumentation in the Center for Biomedical Mass Spectrometry Research at the Medical College of Wisconsin. Tissue sectioning was performed by the Children's Research Institute Histology core lab. Flow cytometry analyses were performed using instrumentation in the Blood Center of Wisconsin Flow Cytometry Core.
Publisher Copyright:
© 2020 Elsevier Ltd
PY - 2020/2
Y1 - 2020/2
N2 - 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.
AB - 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.
KW - Cardiomyocytes
KW - Glycan structures
KW - Heart
KW - Mass spectrometry
KW - Protein glycosylation
KW - hPSC-CM
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UR - http://www.scopus.com/inward/citedby.url?scp=85078130019&partnerID=8YFLogxK
U2 - 10.1016/j.yjmcc.2019.12.012
DO - 10.1016/j.yjmcc.2019.12.012
M3 - Article
C2 - 31972267
AN - SCOPUS:85078130019
VL - 139
SP - 33
EP - 46
JO - Journal of Molecular and Cellular Cardiology
JF - Journal of Molecular and Cellular Cardiology
SN - 0022-2828
ER -