Modification of Lhx2 activity for ex vivo amplification of human iPSC-derived hematopoietic stem/progenitor cells

Kenji Kitajima; Yuna Takahashi; Hikaru Ando; Minako Shingai; Mako Hamasaki; Miyu Tanikawa; Mai Kanokoda; Marino Nakajima; Yasumasa Nishito; Takahiko Hara

doi:10.3389/fcell.2024.1482989

Modification of Lhx2 activity for ex vivo amplification of human iPSC-derived hematopoietic stem/progenitor cells

Front Cell Dev Biol. 2024 Oct 15:12:1482989. doi: 10.3389/fcell.2024.1482989. eCollection 2024.

Authors

Kenji Kitajima¹, Yuna Takahashi^{1

2}, Hikaru Ando^{1

2}, Minako Shingai^{1

2}, Mako Hamasaki^{1

3}, Miyu Tanikawa^{1

2}, Mai Kanokoda^{1

2}, Marino Nakajima^{1

2}, Yasumasa Nishito⁴, Takahiko Hara^{1

2

3}

Affiliations

¹ Stem Cell Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.
² Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
³ Graduate School of Science, Department of Biological Science, Tokyo Metropolitan University, Tokyo, Japan.
⁴ Center for Basic Technology Research, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan.

Abstract

Hematopoietic stem cells (HSCs) obtained from patient-derived human induced pluripotent stem cells (iPSCs) are a promising tool for curing various hematological disorders. We previously demonstrated that enforced expression of the LIM-homeobox transcription factor Lhx2, which is essential for mouse embryonic hematopoiesis, leads to generation of engraftable and expandable hematopoietic stem cells (HSCs) from mouse iPSCs. However, it remained unknown whether Lhx2 can induce HSCs from human iPSCs. Here, we investigated the effect of Lhx2 overexpression on hematopoietic differentiation of human iPSCs. Unexpectedly, Lhx2 severely inhibited proliferation of human iPSC-derived hematopoietic cells. Thus, Lhx2 exhibited differential effects on mouse and human hematopoietic cells. Further studies implied that the inhibitory effect of Lhx2 on human iPSC-derived hematopoietic cells was due to insufficient transcriptional activation ability. Therefore, we modified Lhx2 to strengthen its activity as a transcriptional activator. This modified Lhx2 could induce ex vivo amplification of human iPSC-derived hematopoietic stem/progenitor cells (HSPCs). We believe that these findings will facilitate the development of a method to efficiently produce HSCs from human iPSCs.

Keywords: HSPCs; Lhx2; differentiation; ex vivo amplification; homeobox; iPSCs; organoid.

Grants and funding

The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (17K09911 to KK and 16H04728 to KK and TH).