TY - JOUR
T1 - KLF1 mutation E325K induces cell cycle arrest in erythroid cells differentiated from congenital dyserythropoietic anemia patient-specific induced pluripotent stem cells
AU - Kohara, Hiroshi
AU - Utsugisawa, Taiju
AU - Sakamoto, Chika
AU - Hirose, Lisa
AU - Ogawa, Yoshie
AU - Ogura, Hiromi
AU - Sugawara, Ai
AU - Liao, Jiyuan
AU - Aoki, Takako
AU - Iwasaki, Takuya
AU - Asai, Takayoshi
AU - Doisaki, Sayoko
AU - Okuno, Yusuke
AU - Muramatsu, Hideki
AU - Abe, Takaaki
AU - Kurita, Ryo
AU - Miyamoto, Shohei
AU - Sakuma, Tetsushi
AU - Shiba, Masayuki
AU - Yamamoto, Takashi
AU - Ohga, Shouichi
AU - Yoshida, Kenichi
AU - Ogawa, Seishi
AU - Ito, Etsuro
AU - Kojima, Seiji
AU - Kanno, Hitoshi
AU - Tani, Kenzaburo
N1 - Funding Information:
We thank the CDA patient who participated in this study; the Flow Cytometry Core Facility and Center for Stem Cell Biology and Regenerative Medicine at The Institute of Medical Science, University of Tokyo, for assistance with instrumentation; Dr. Satoshi Yamazaki for technical assistance with the teratoma assay; and Dr. Hiroaki Ono and Dr. Hidetoshi Takada for providing helpful comments for blood collection. This work was supported by the Japan Agency for Medical Research and Development (AMED), SBI Pharmaceutical, and Shinnihonseiyaku. Hiroshi Kohara, Shohei Miyamoto and Kenzaburo Tani are supported by grants from Shinnihonseiyaku Company, SBI Pharmaceutical. Co, Ltd and neopharm Japan Co. Ltd. The terms of this arrangement have been reviewed and approved by the University of Tokyo in accordance with its conflict of interest policies. The remaining authors declare no competing financial interests.
Funding Information:
Hiroshi Kohara, Shohei Miyamoto and Kenzaburo Tani are supported by grants from Shinnihonseiyaku Company, SBI Pharmaceutical. Co, Ltd and neopharm Japan Co., Ltd. The terms of this arrangement have been reviewed and approved by the University of Tokyo in accordance with its conflict of interest policies. The remaining authors declare no competing financial interests.
Funding Information:
We thank the CDA patient who participated in this study; the Flow Cytometry Core Facility and Center for Stem Cell Biology and Regenerative Medicine at The Institute of Medical Science, University of Tokyo, for assistance with instrumentation; Dr. Satoshi Yamazaki for technical assistance with the teratoma assay; and Dr. Hiroaki Ono and Dr. Hidetoshi Takada for providing helpful comments for blood collection. This work was supported by the Japan Agency for Medical Research and Development (AMED), SBI Pharmaceutical, and Shinnihonseiyaku.
Publisher Copyright:
© 2019
PY - 2019/5
Y1 - 2019/5
N2 - Krüppel-like factor 1 (KLF1), a transcription factor controlling definitive erythropoiesis, is involved in sequential control of terminal cell division and enucleation via fine regulation of key cell cycle regulator gene expression in erythroid lineage cells. Type IV congenital dyserythropoietic anemia (CDA)is caused by a monoallelic mutation at the second zinc finger of KLF1 (c.973G>A, p.E325K). We recently diagnosed a female patient with type IV CDA with the identical missense mutation. To understand the mechanism underlying the dyserythropoiesis caused by the mutation, we generated induced pluripotent stem cells (iPSCs)from the CDA patient (CDA-iPSCs). The erythroid cells that differentiated from CDA-iPSCs (CDA-erythroid cells)displayed multinucleated morphology, absence of CD44, and dysregulation of the KLF1 target gene expression. In addition, uptake of bromodeoxyuridine by CDA-erythroid cells was significantly decreased at the CD235a+/CD71+ stage, and microarray analysis revealed that cell cycle regulator genes were dysregulated, with increased expression of negative regulators such as CDKN2C and CDKN2A. Furthermore, inducible expression of the KLF1 E325K, but not the wild-type KLF1, caused a cell cycle arrest at the G1 phase in CDA-erythroid cells. Microarray analysis of CDA-erythroid cells and real-time polymerase chain reaction analysis of the KLF1 E325K inducible expression system also revealed altered expression of several KLF1 target genes including erythrocyte membrane protein band 4.1 (EPB41), EPB42, glutathione disulfide reductase (GSR), glucose phosphate isomerase (GPI), and ATPase phospholipid transporting 8A1 (ATP8A1). Our data indicate that the E325K mutation in KLF1 is associated with disruption of transcriptional control of cell cycle regulators in association with erythroid membrane or enzyme abnormalities, leading to dyserythropoiesis.
AB - Krüppel-like factor 1 (KLF1), a transcription factor controlling definitive erythropoiesis, is involved in sequential control of terminal cell division and enucleation via fine regulation of key cell cycle regulator gene expression in erythroid lineage cells. Type IV congenital dyserythropoietic anemia (CDA)is caused by a monoallelic mutation at the second zinc finger of KLF1 (c.973G>A, p.E325K). We recently diagnosed a female patient with type IV CDA with the identical missense mutation. To understand the mechanism underlying the dyserythropoiesis caused by the mutation, we generated induced pluripotent stem cells (iPSCs)from the CDA patient (CDA-iPSCs). The erythroid cells that differentiated from CDA-iPSCs (CDA-erythroid cells)displayed multinucleated morphology, absence of CD44, and dysregulation of the KLF1 target gene expression. In addition, uptake of bromodeoxyuridine by CDA-erythroid cells was significantly decreased at the CD235a+/CD71+ stage, and microarray analysis revealed that cell cycle regulator genes were dysregulated, with increased expression of negative regulators such as CDKN2C and CDKN2A. Furthermore, inducible expression of the KLF1 E325K, but not the wild-type KLF1, caused a cell cycle arrest at the G1 phase in CDA-erythroid cells. Microarray analysis of CDA-erythroid cells and real-time polymerase chain reaction analysis of the KLF1 E325K inducible expression system also revealed altered expression of several KLF1 target genes including erythrocyte membrane protein band 4.1 (EPB41), EPB42, glutathione disulfide reductase (GSR), glucose phosphate isomerase (GPI), and ATPase phospholipid transporting 8A1 (ATP8A1). Our data indicate that the E325K mutation in KLF1 is associated with disruption of transcriptional control of cell cycle regulators in association with erythroid membrane or enzyme abnormalities, leading to dyserythropoiesis.
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U2 - 10.1016/j.exphem.2019.03.001
DO - 10.1016/j.exphem.2019.03.001
M3 - Article
C2 - 30876823
AN - SCOPUS:85063763048
SN - 0301-472X
VL - 73
SP - 25-37.e8
JO - Experimental Hematology
JF - Experimental Hematology
ER -
