Single-cell transcriptome analysis reveals CD34 as a marker of human sinoatrial node pacemaker cardiomyocytes

Amos A Lim; Delaram Pouyabahar; Mishal Ashraf; Kate Huang; Michelle Lohbihler; Brandon M Murareanu; Matthew L Chang; Maggie Kwan; Faisal J Alibhai; Thinh Tran; Amine Mazine; Michael A Laflamme; Gary D Bader; Zachary Laksman; Stephanie Protze

doi:10.1038/s41467-024-54337-4

Single-cell transcriptome analysis reveals CD34 as a marker of human sinoatrial node pacemaker cardiomyocytes

Nat Commun. 2024 Nov 27;15(1):10206. doi: 10.1038/s41467-024-54337-4.

Authors

Amos A Lim^{1

2}, Delaram Pouyabahar^{2

3}, Mishal Ashraf^{4

5}, Kate Huang^{4

6}, Michelle Lohbihler^{1

2}, Brandon M Murareanu^{1

2}, Matthew L Chang^{1

2}, Maggie Kwan¹, Faisal J Alibhai¹, Thinh Tran^{2

3

7}, Amine Mazine^{1

8

9}, Michael A Laflamme^{1

10}, Gary D Bader^{2

3

11

12

13}, Zachary Laksman^{4

5

6

14}, Stephanie Protze^{15

16}

Affiliations

¹ McEwen Stem Cell Institute, University Health Network, Toronto, ON, Canada.
² Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.
³ The Donnelly Centre, University of Toronto, Toronto, ON, Canada.
⁴ Centre for Heart and Lung Innovation, University of British Columbia and St. Paul's Hospital, Vancouver, BC, Canada.
⁵ School of Biomedical Engineering, University of British Columbia, Vancouver, BC, Canada.
⁶ Experimental Medicine Program, University of British Columbia, Vancouver, BC, Canada.
⁷ Marie-Josée and Henry R. Kravis Center for Molecular Oncology, Memorial Sloan Kettering Cancer Center, New York, NY, USA.
⁸ Institute of Biomedical Engineering, University of Toronto, Toronto, ON, Canada.
⁹ Division of Cardiac Surgery, Department of Surgery, University of Toronto, Toronto, ON, Canada.
¹⁰ Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada.
¹¹ Department of Computer Science, University of Toronto, Toronto, ON, Canada.
¹² Princess Margaret Cancer Centre, University Health Network, Toronto, ON, Canada.
¹³ Lunenfeld-Tanenbaum Research Institute, Sinai Health System, Toronto, ON, Canada.
¹⁴ Department of Medicine, University of British Columbia, Vancouver, BC, Canada.
¹⁵ McEwen Stem Cell Institute, University Health Network, Toronto, ON, Canada. [email protected].
¹⁶ Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada. [email protected].

Abstract

The sinoatrial node regulates the heart rate throughout life. Failure of this primary pacemaker results in life-threatening, slow heart rhythm. Despite its critical function, the cellular and molecular composition of the human sinoatrial node is not resolved. Particularly, no cell surface marker to identify and isolate sinoatrial node pacemaker cells has been reported. Here we use single-nuclei/cell RNA sequencing of fetal and human pluripotent stem cell-derived sinoatrial node cells to reveal that they consist of three subtypes of pacemaker cells: Core Pacemaker, Sinus Venosus, and Transitional Cells. Our study identifies a host of sinoatrial node pacemaker markers including MYH11, BMP4, and the cell surface antigen CD34. We demonstrate that sorting for CD34⁺ cells from stem cell differentiation cultures enriches for sinoatrial node cells exhibiting a functional pacemaker phenotype. This sinoatrial node pacemaker cell surface marker is highly valuable for stem cell-based disease modeling, drug discovery, cell replacement therapies, and the targeted delivery of therapeutics to sinoatrial node cells in vivo using antibody-drug conjugates.

MeSH terms

Antigens, CD34* / metabolism
Biomarkers / metabolism
Cell Differentiation*
Gene Expression Profiling / methods
Humans
Induced Pluripotent Stem Cells / cytology
Induced Pluripotent Stem Cells / metabolism
Myocytes, Cardiac* / cytology
Myocytes, Cardiac* / metabolism
Pluripotent Stem Cells / cytology
Pluripotent Stem Cells / metabolism
Single-Cell Analysis* / methods
Single-Cell Gene Expression Analysis
Sinoatrial Node* / cytology
Sinoatrial Node* / metabolism
Transcriptome

Substances

Antigens, CD34
Biomarkers

Abstract

MeSH terms

Substances

Grants and funding