TY - JOUR
T1 - A single cell transcriptional roadmap of human pacemaker cell differentiation
AU - Wiesinger, Alexandra
AU - Li, Jiuru
AU - Fokkert, Lianne
AU - Bakker, Priscilla
AU - Verkerk, Arie O.
AU - Christoffels, Vincent M.
AU - Boink, Gerard J. J.
AU - Devalla, Harsha D.
N1 - Funding Information: We thank Berend Hooibrink from the Flow Cytometry facility, Department of Medical Biology, for assistance with cell sorting, Corrie de Gier-de Vries, Department of Medical Biology, for help with histology of mouse hearts and Likhitha Puliyadi, Department of Medical Biology for assistance with RT-qPCR. We also gratefully acknowledge funding from the European Research council starting grant 714866 and associated proof-of-concept grant 899422, Health Holland LentiPace II, Horizon 2020 Eurostars (E114245 and E115484), Dutch Research Council Open Technology Program 18485 to GJJB; Netherlands Organization for Health Research and Development (ZonMW), ZonMW TOP 40-00812-98-17061 to VMC, ZonMW and the Dutch Heart foundation MKMD grant 114021512 and Dutch Heart Foundation Dekker fellowship 2020T023 to HDD. Publisher Copyright: © Wiesinger et al.
PY - 2022/10/11
Y1 - 2022/10/11
N2 - Each heartbeat is triggered by the sinoatrial node (SAN), the primary pacemaker of the heart. Studies in animal models have revealed that pacemaker cells share a common progenitor with the (pro)epicardium, and that the pacemaker cardiomyocytes further diversify into 'transitional', 'tail', and 'head' subtypes. However, the underlying molecular mechanisms, especially of human pacemaker cell development, are poorly understood. Here, we performed single cell RNA sequencing (scRNA-seq) and trajectory inference on human induced pluripotent stem cells (hiPSCs) differentiating to SAN-like cardiomyocytes (SANCMs) to construct a roadmap of transcriptional changes and lineage decisions. In differentiated SANCM, we identified distinct clusters that closely resemble different subpopulations of the in vivo SAN. Moreover, the presence of a side population of proepicardial cells suggested their shared ontogeny with SANCM, as also reported in vivo. Our results demonstrate that the divergence of SANCM and proepicardial lineages is determined by WNT signaling. Furthermore, we uncovered roles for TGFβ and WNT signaling in the branching of transitional and head SANCM subtypes, respectively. These findings provide new insights into the molecular processes involved in human pacemaker cell differentiation, opening new avenues for complex disease modeling in vitro and inform approaches for cell therapy-based regeneration of the SAN.
AB - Each heartbeat is triggered by the sinoatrial node (SAN), the primary pacemaker of the heart. Studies in animal models have revealed that pacemaker cells share a common progenitor with the (pro)epicardium, and that the pacemaker cardiomyocytes further diversify into 'transitional', 'tail', and 'head' subtypes. However, the underlying molecular mechanisms, especially of human pacemaker cell development, are poorly understood. Here, we performed single cell RNA sequencing (scRNA-seq) and trajectory inference on human induced pluripotent stem cells (hiPSCs) differentiating to SAN-like cardiomyocytes (SANCMs) to construct a roadmap of transcriptional changes and lineage decisions. In differentiated SANCM, we identified distinct clusters that closely resemble different subpopulations of the in vivo SAN. Moreover, the presence of a side population of proepicardial cells suggested their shared ontogeny with SANCM, as also reported in vivo. Our results demonstrate that the divergence of SANCM and proepicardial lineages is determined by WNT signaling. Furthermore, we uncovered roles for TGFβ and WNT signaling in the branching of transitional and head SANCM subtypes, respectively. These findings provide new insights into the molecular processes involved in human pacemaker cell differentiation, opening new avenues for complex disease modeling in vitro and inform approaches for cell therapy-based regeneration of the SAN.
KW - cardiac differentiation
KW - developmental biology
KW - human
KW - iPSC
KW - pacemaker cell
KW - regenerative medicine
KW - scRNA-seq
KW - sinoatrial node
KW - stem cells
KW - trajectory inference
UR - http://www.scopus.com/inward/record.url?scp=85139532223&partnerID=8YFLogxK
U2 - https://doi.org/10.7554/eLife.76781
DO - https://doi.org/10.7554/eLife.76781
M3 - Article
C2 - 36217819
SN - 2050-084X
VL - 11
JO - eLife
JF - eLife
M1 - e76781
ER -