Hematopoiesis by iPSC-derived hematopoietic stem cells of aplastic anemia that escape cytotoxic T-cell attack

J. Luis Espinoza, Mahmoud I. Elbadry, Kazuhisa Chonabayashi, Yoshinori Yoshida, Takamasa Katagiri, Kenichi Harada, Noriharu Nakagawa, Yoshitaka Zaimoku, Tatsuya Imi, Hiroyuki Takamatsu, Tatsuhiko Ozawa, Hiroyuki Maruyama, Hassan A. Hassanein, Amal Khalifa A. Noreldin, Katsuto Takenaka, Koichi Akashi, Hiroshi Hamana, Hiroyuki Kishi, Yoshiki Akatsuka, Shinji Nakao

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Hematopoietic stem cells (HSCs) that lack HLA-class I alleles as a result of copy-number neutral loss of heterozygosity of the short arm of chromosome 6 (6pLOH) or HLA allelic mutations often constitute hematopoiesis in patients with acquired aplastic anemia (AA), but the precise mechanisms underlying clonal hematopoiesis induced by these HLA-lacking (HLA-) HSCs remain unknown. To address this issue, we generated induced pluripotent stem cells (iPSCs) from an AA patient who possessed HLA-B4002-lacking (B4002-) leukocytes. Three different iPSC clones (wild-type [WT], 6pLOH+, and B∗40:02-mutant) were established from the patient's monocytes. Three-week cultures of the iPSCs in the presence of various growth factors produced hematopoietic cells that make up 50% to 70%of the CD34+ cells of each phenotype. When 106 iPSC-derived CD34+ (iCD34+) cells with the 3 different genotypes were injected into the femoral bone of C57BL/6.Rag2 mice, 2.1% to 7.3% human multilineage CD45+ cells of each HLA phenotype were detected in the bone marrow, spleen, and peripheral blood of themice at 9 to 12weeks after the injection, with no significant difference in the human: mouse chimerism ratio among the 3 groups. Stimulation of the patient's CD8+ T cellswith the WTiCD34+ cells generated a cytotoxic T lymphocyte (CTL) line capable of killingWT iCD34+ cells but not B4002- iCD34+ cells. These data suggest that B4002- iCD34+ cells show a repopulating ability similar to that of WT iCD34+ cells when autologous T cells are absent and CTL precursors capable of selectively killing WT HSCs are present in the patient's peripheral blood.

Original languageEnglish
Pages (from-to)390-400
Number of pages11
JournalBlood Advances
Volume2
Issue number4
DOIs
Publication statusPublished - 27-02-2018

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Induced Pluripotent Stem Cells
Aplastic Anemia
Hematopoiesis
Hematopoietic Stem Cells
T-Lymphocytes
Cytotoxic T-Lymphocytes
Hematopoietic Cell Growth Factors
Phenotype
Chimerism
Chromosomes, Human, Pair 6
Loss of Heterozygosity
Thigh
Monocytes
Leukocytes
Spleen
Clone Cells
Bone Marrow
Alleles
Genotype
Bone and Bones

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Espinoza, J. L., Elbadry, M. I., Chonabayashi, K., Yoshida, Y., Katagiri, T., Harada, K., ... Nakao, S. (2018). Hematopoiesis by iPSC-derived hematopoietic stem cells of aplastic anemia that escape cytotoxic T-cell attack. Blood Advances, 2(4), 390-400. https://doi.org/10.1182/bloodadvances.2017013342
Espinoza, J. Luis ; Elbadry, Mahmoud I. ; Chonabayashi, Kazuhisa ; Yoshida, Yoshinori ; Katagiri, Takamasa ; Harada, Kenichi ; Nakagawa, Noriharu ; Zaimoku, Yoshitaka ; Imi, Tatsuya ; Takamatsu, Hiroyuki ; Ozawa, Tatsuhiko ; Maruyama, Hiroyuki ; Hassanein, Hassan A. ; Noreldin, Amal Khalifa A. ; Takenaka, Katsuto ; Akashi, Koichi ; Hamana, Hiroshi ; Kishi, Hiroyuki ; Akatsuka, Yoshiki ; Nakao, Shinji. / Hematopoiesis by iPSC-derived hematopoietic stem cells of aplastic anemia that escape cytotoxic T-cell attack. In: Blood Advances. 2018 ; Vol. 2, No. 4. pp. 390-400.
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abstract = "Hematopoietic stem cells (HSCs) that lack HLA-class I alleles as a result of copy-number neutral loss of heterozygosity of the short arm of chromosome 6 (6pLOH) or HLA allelic mutations often constitute hematopoiesis in patients with acquired aplastic anemia (AA), but the precise mechanisms underlying clonal hematopoiesis induced by these HLA-lacking (HLA-) HSCs remain unknown. To address this issue, we generated induced pluripotent stem cells (iPSCs) from an AA patient who possessed HLA-B4002-lacking (B4002-) leukocytes. Three different iPSC clones (wild-type [WT], 6pLOH+, and B∗40:02-mutant) were established from the patient's monocytes. Three-week cultures of the iPSCs in the presence of various growth factors produced hematopoietic cells that make up 50{\%} to 70{\%}of the CD34+ cells of each phenotype. When 106 iPSC-derived CD34+ (iCD34+) cells with the 3 different genotypes were injected into the femoral bone of C57BL/6.Rag2 mice, 2.1{\%} to 7.3{\%} human multilineage CD45+ cells of each HLA phenotype were detected in the bone marrow, spleen, and peripheral blood of themice at 9 to 12weeks after the injection, with no significant difference in the human: mouse chimerism ratio among the 3 groups. Stimulation of the patient's CD8+ T cellswith the WTiCD34+ cells generated a cytotoxic T lymphocyte (CTL) line capable of killingWT iCD34+ cells but not B4002- iCD34+ cells. These data suggest that B4002- iCD34+ cells show a repopulating ability similar to that of WT iCD34+ cells when autologous T cells are absent and CTL precursors capable of selectively killing WT HSCs are present in the patient's peripheral blood.",
author = "Espinoza, {J. Luis} and Elbadry, {Mahmoud I.} and Kazuhisa Chonabayashi and Yoshinori Yoshida and Takamasa Katagiri and Kenichi Harada and Noriharu Nakagawa and Yoshitaka Zaimoku and Tatsuya Imi and Hiroyuki Takamatsu and Tatsuhiko Ozawa and Hiroyuki Maruyama and Hassanein, {Hassan A.} and Noreldin, {Amal Khalifa A.} and Katsuto Takenaka and Koichi Akashi and Hiroshi Hamana and Hiroyuki Kishi and Yoshiki Akatsuka and Shinji Nakao",
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Espinoza, JL, Elbadry, MI, Chonabayashi, K, Yoshida, Y, Katagiri, T, Harada, K, Nakagawa, N, Zaimoku, Y, Imi, T, Takamatsu, H, Ozawa, T, Maruyama, H, Hassanein, HA, Noreldin, AKA, Takenaka, K, Akashi, K, Hamana, H, Kishi, H, Akatsuka, Y & Nakao, S 2018, 'Hematopoiesis by iPSC-derived hematopoietic stem cells of aplastic anemia that escape cytotoxic T-cell attack', Blood Advances, vol. 2, no. 4, pp. 390-400. https://doi.org/10.1182/bloodadvances.2017013342

Hematopoiesis by iPSC-derived hematopoietic stem cells of aplastic anemia that escape cytotoxic T-cell attack. / Espinoza, J. Luis; Elbadry, Mahmoud I.; Chonabayashi, Kazuhisa; Yoshida, Yoshinori; Katagiri, Takamasa; Harada, Kenichi; Nakagawa, Noriharu; Zaimoku, Yoshitaka; Imi, Tatsuya; Takamatsu, Hiroyuki; Ozawa, Tatsuhiko; Maruyama, Hiroyuki; Hassanein, Hassan A.; Noreldin, Amal Khalifa A.; Takenaka, Katsuto; Akashi, Koichi; Hamana, Hiroshi; Kishi, Hiroyuki; Akatsuka, Yoshiki; Nakao, Shinji.

In: Blood Advances, Vol. 2, No. 4, 27.02.2018, p. 390-400.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Hematopoiesis by iPSC-derived hematopoietic stem cells of aplastic anemia that escape cytotoxic T-cell attack

AU - Espinoza, J. Luis

AU - Elbadry, Mahmoud I.

AU - Chonabayashi, Kazuhisa

AU - Yoshida, Yoshinori

AU - Katagiri, Takamasa

AU - Harada, Kenichi

AU - Nakagawa, Noriharu

AU - Zaimoku, Yoshitaka

AU - Imi, Tatsuya

AU - Takamatsu, Hiroyuki

AU - Ozawa, Tatsuhiko

AU - Maruyama, Hiroyuki

AU - Hassanein, Hassan A.

AU - Noreldin, Amal Khalifa A.

AU - Takenaka, Katsuto

AU - Akashi, Koichi

AU - Hamana, Hiroshi

AU - Kishi, Hiroyuki

AU - Akatsuka, Yoshiki

AU - Nakao, Shinji

PY - 2018/2/27

Y1 - 2018/2/27

N2 - Hematopoietic stem cells (HSCs) that lack HLA-class I alleles as a result of copy-number neutral loss of heterozygosity of the short arm of chromosome 6 (6pLOH) or HLA allelic mutations often constitute hematopoiesis in patients with acquired aplastic anemia (AA), but the precise mechanisms underlying clonal hematopoiesis induced by these HLA-lacking (HLA-) HSCs remain unknown. To address this issue, we generated induced pluripotent stem cells (iPSCs) from an AA patient who possessed HLA-B4002-lacking (B4002-) leukocytes. Three different iPSC clones (wild-type [WT], 6pLOH+, and B∗40:02-mutant) were established from the patient's monocytes. Three-week cultures of the iPSCs in the presence of various growth factors produced hematopoietic cells that make up 50% to 70%of the CD34+ cells of each phenotype. When 106 iPSC-derived CD34+ (iCD34+) cells with the 3 different genotypes were injected into the femoral bone of C57BL/6.Rag2 mice, 2.1% to 7.3% human multilineage CD45+ cells of each HLA phenotype were detected in the bone marrow, spleen, and peripheral blood of themice at 9 to 12weeks after the injection, with no significant difference in the human: mouse chimerism ratio among the 3 groups. Stimulation of the patient's CD8+ T cellswith the WTiCD34+ cells generated a cytotoxic T lymphocyte (CTL) line capable of killingWT iCD34+ cells but not B4002- iCD34+ cells. These data suggest that B4002- iCD34+ cells show a repopulating ability similar to that of WT iCD34+ cells when autologous T cells are absent and CTL precursors capable of selectively killing WT HSCs are present in the patient's peripheral blood.

AB - Hematopoietic stem cells (HSCs) that lack HLA-class I alleles as a result of copy-number neutral loss of heterozygosity of the short arm of chromosome 6 (6pLOH) or HLA allelic mutations often constitute hematopoiesis in patients with acquired aplastic anemia (AA), but the precise mechanisms underlying clonal hematopoiesis induced by these HLA-lacking (HLA-) HSCs remain unknown. To address this issue, we generated induced pluripotent stem cells (iPSCs) from an AA patient who possessed HLA-B4002-lacking (B4002-) leukocytes. Three different iPSC clones (wild-type [WT], 6pLOH+, and B∗40:02-mutant) were established from the patient's monocytes. Three-week cultures of the iPSCs in the presence of various growth factors produced hematopoietic cells that make up 50% to 70%of the CD34+ cells of each phenotype. When 106 iPSC-derived CD34+ (iCD34+) cells with the 3 different genotypes were injected into the femoral bone of C57BL/6.Rag2 mice, 2.1% to 7.3% human multilineage CD45+ cells of each HLA phenotype were detected in the bone marrow, spleen, and peripheral blood of themice at 9 to 12weeks after the injection, with no significant difference in the human: mouse chimerism ratio among the 3 groups. Stimulation of the patient's CD8+ T cellswith the WTiCD34+ cells generated a cytotoxic T lymphocyte (CTL) line capable of killingWT iCD34+ cells but not B4002- iCD34+ cells. These data suggest that B4002- iCD34+ cells show a repopulating ability similar to that of WT iCD34+ cells when autologous T cells are absent and CTL precursors capable of selectively killing WT HSCs are present in the patient's peripheral blood.

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