Selective vulnerability of human-induced pluripotent stem cells to dihydroorotate dehydrogenase inhibition during mesenchymal stem/stromal cell purification

Ziadoon Al-Akashi; Denise Zujur; Daisuke Kamiya; Tomohisa Kato Jr; Toru Kondo; Makoto Ikeya

doi:10.3389/fcell.2023.1089945

Selective vulnerability of human-induced pluripotent stem cells to dihydroorotate dehydrogenase inhibition during mesenchymal stem/stromal cell purification

Front Cell Dev Biol. 2023 Feb 6:11:1089945. doi: 10.3389/fcell.2023.1089945. eCollection 2023.

Authors

Ziadoon Al-Akashi¹, Denise Zujur¹, Daisuke Kamiya^{1

2}, Tomohisa Kato Jr³, Toru Kondo⁴, Makoto Ikeya^{1

2}

Affiliations

¹ Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan.
² Takeda-CiRA Joint Program, Fujisawa, Kanagawa, Japan.
³ Medical Research Institute, Kanazawa Medical University, Kanazawa, Japan.
⁴ Institute for Genetic Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.

Abstract

The use of induced mesenchymal stem/stromal cells (iMSCs) derived from human induced pluripotent stem cells (hiPSCs) in regenerative medicine involves the risk of teratoma formation due to hiPSCs contamination in iMSCs. Therefore, eradicating the remaining undifferentiated hiPSCs is crucial for the effectiveness of the strategy. The present study demonstrates the Brequinar (BRQ)-induced inhibition of dihydroorotate dehydrogenase (DHODH), a key enzyme in de novo pyrimidine biosynthesis, selectively induces apoptosis, cell cycle arrest, and differentiation; furthermore, it promotes transcriptional changes and prevents the growth of 3-dimensional hiPSC aggregates. Contrastingly, BRQ-treated iMSCs showed no changes in survival, differentiation potential, or gene expression. The results suggest that BRQ is a potential agent for the effective purification of iMSCs from a mixed population of iMSCs and hiPSCs, which is a crucial step in successful iMSC-based therapy.

Keywords: brequinar (BRQ); dihydroorotate dehydrogenase (DHODH); induced mesenchymal stromal cells (iMSCs); induced pluripotent stem cells (hiPSC); mesenchymal stem/stromal cells (MSCs); pyrimidine biosynthesis; survival.

Grants and funding

This research was supported by the Japan Agency for Medical Research and Development (AMED) under Grant Number JP15bm0104001 and JP22bm1123006, grants-in-aid for scientific research from the Japan Society for the Promotion of Science (JSPS) (#22H03275, #22H03204), the iPS Cell Research Fund, KOSÉ Cosmetology Research Foundation, and Takeda Pharmaceutical Company Limited to MI. This work was also supported by JST SPRING (grant number JPMJSP2110) to ZA. This study was carried out under the Joint Research Program of the Institute for Genetic Medicine, Hokkaido University. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article or the decision to submit it for publication.