Expression dynamics of HAND1/2 in in vitro human cardiomyocyte differentiation

Chikako Okubo; Megumi Narita; Azusa Inagaki; Misato Nishikawa; Akitsu Hotta; Shinya Yamanaka; Yoshinori Yoshida

doi:10.1016/j.stemcr.2021.06.014

Expression dynamics of HAND1/2 in in vitro human cardiomyocyte differentiation

Stem Cell Reports. 2021 Aug 10;16(8):1906-1922. doi: 10.1016/j.stemcr.2021.06.014. Epub 2021 Jul 22.

Authors

Chikako Okubo¹, Megumi Narita¹, Azusa Inagaki¹, Misato Nishikawa¹, Akitsu Hotta², Shinya Yamanaka³, Yoshinori Yoshida⁴

Affiliations

¹ Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
² Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan.
³ Department of Life Science Frontiers, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan; Gladstone Institute of Cardiovascular Disease, Gladstone Institutes, San Francisco, CA 94158, USA.
⁴ Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto 606-8507, Japan. Electronic address: yoshinor@cira.kyoto-u.ac.jp.

Abstract

Hand1 and Hand2 are transcriptional factors, and knockout mice of these genes show left and right ventricular hypoplasia, respectively. However, their function and expression in human cardiogenesis are not well studied. To delineate their expressions and assess their functions in human cardiomyocytes (CMs) in vitro, we established two triple-reporter human induced pluripotent stem cell lines that express HAND1^mCherry, HAND2^EGFP and either MYH6-driven iRFP670 or tagBFP constitutively and investigated their expression dynamics during cardiac differentiation. On day 5 of the differentiation, HAND1 expression marked cardiac progenitor cells. We profiled the CM subpopulations on day 20 with RNA sequencing and found that mCherry+ CMs showed higher proliferative ability than mCherry- CMs and identified a gene network of LEF1, HAND1, and HAND2 to regulate proliferation in CMs. Finally, we identified CD105 as a surface marker of highly proliferative CMs.

Keywords: HAND1; HAND2; cardiomyocyte; development; pluripotent stem cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Basic Helix-Loop-Helix Transcription Factors / genetics*
Basic Helix-Loop-Helix Transcription Factors / metabolism
Cell Cycle / genetics
Cell Differentiation / genetics*
Cell Proliferation / genetics
Cells, Cultured
Gene Regulatory Networks / genetics
Humans
Induced Pluripotent Stem Cells / metabolism
Luminescent Proteins / genetics
Luminescent Proteins / metabolism
Myocytes, Cardiac / cytology
Myocytes, Cardiac / metabolism*
RNA Interference
RNA-Seq / methods
Time Factors
Transcriptome / genetics*

Substances

Basic Helix-Loop-Helix Transcription Factors
HAND2 protein, human
Luminescent Proteins
helix-loop-helix protein, eHAND