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Non-Lethal Ionizing Radiation Promotes Aging-Like Phenotypic Changes of Human Hematopoietic Stem and Progenitor Cells in Humanized Mice.

Wang C, Oshima M, Sashida G, Tomioka T, Hasegawa N, Mochizuki-Kashio M, Nakajima-Takagi Y, Kusunoki Y, Kyoizumi S, Imai K, Nakachi K, Iwama A - PLoS ONE (2015)

Bottom Line: Of interest, non-lethal radiation compromised contribution of HSPCs to the peripheral blood cells, particularly to CD19+ B lymphocytes, which resulted in myeloid-biased repopulation.Co-culture of limiting numbers of CD34+ HSPCs with stromal cells revealed that the frequency of B cell-producing CD34+ HSPCs at 2 weeks post-irradiation was reduced more than 10-fold.Given that compromised repopulating capacity and myeloid-biased differentiation are representative phenotypes of aged HSCs, our findings indicate that non-lethal ionizing radiation is one of the critical external stresses that promote aging of human HSPCs in the bone marrow niche.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Medicine, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan; College of Life Sciences, Inner Mongolia University, Hohhot 010021, China.

ABSTRACT
Precise understanding of radiation effects is critical to develop new modalities for the prevention and treatment of radiation-induced damage. We previously reported that non-lethal doses of X-ray irradiation induce DNA damage in human hematopoietic stem and progenitor cells (HSPCs) reconstituted in NOD/Shi-scid IL2rγ (NOG) immunodeficient mice and severely compromise their repopulating capacity. In this study, we analyzed in detail the functional changes in human HSPCs in NOG mice following non-lethal radiation. We transplanted cord blood CD34+ HSPCs into NOG mice. At 12 weeks post-transplantation, the recipients were irradiated with 0, 0.5, or 1.0 Gy. At 2 weeks post-irradiation, human CD34+ HSPCs recovered from the primary recipient mice were transplanted into secondary recipients. CD34+ HSPCs from irradiated mice showed severely impaired reconstitution capacity in the secondary recipient mice. Of interest, non-lethal radiation compromised contribution of HSPCs to the peripheral blood cells, particularly to CD19+ B lymphocytes, which resulted in myeloid-biased repopulation. Co-culture of limiting numbers of CD34+ HSPCs with stromal cells revealed that the frequency of B cell-producing CD34+ HSPCs at 2 weeks post-irradiation was reduced more than 10-fold. Furthermore, the key B-cell regulator genes such as IL-7R and EBF1 were downregulated in HSPCs upon 0.5 Gy irradiation. Given that compromised repopulating capacity and myeloid-biased differentiation are representative phenotypes of aged HSCs, our findings indicate that non-lethal ionizing radiation is one of the critical external stresses that promote aging of human HSPCs in the bone marrow niche.

No MeSH data available.


Related in: MedlinePlus

Early effects of irradiation on human HSPCs in humanized mice.(A) Changes in chimerism of human hematopoietic cells in the PB after irradiation. At 12 weeks post-irradiation, the mice were randomly grouped into three categories and irradiated with 0, 0.5 or 1.0 Gy for each group. At 2 weeks post-irradiation, mice were sacrificed to evaluate early effects of irradiation. White blood cell (WBC) counts in the PB (left panel) and chimerism of CD45+ human (middle panel) and mouse (right panel) hematopoietic cells in the PB are presented as scatter diagrams with median values (bars) (n = 9 each). PB data of NOG mice just before the irradiation (Pre) are presented as controls (n = 27). (B) The percentages of CD11b+ myeloid cells, CD19+ B cells, and CD3+ T cells in CD45+ human hematopoietic cells in the PB of NOG mice just before the irradiation (Pre, n = 27) and at 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (C) Total BM cell numbers (left panel) and chimerism of CD45+ human (middle) and mouse (right) hematopoietic cells in the BM just before the irradiation (Pre, n = 6) and at 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (D) Absolute numbers of CD34+CD38- HSCs, CD34+CD38+ HPCs, CMPs and LMPPs in the BM (bilateral femurs and tibiae) just before the irradiation (Pre, n = 6) and 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (E) Numbers of colony-forming cells included in 1,000 CD34+ human HSPCs recovered at 2 weeks after irradiation (0–1.0 Gy, n = 3 each). Bars in scatter diagrams indicate median values. P-values for trend were calculated by nonparametric Jonckheere-Terpstra test.
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pone.0132041.g002: Early effects of irradiation on human HSPCs in humanized mice.(A) Changes in chimerism of human hematopoietic cells in the PB after irradiation. At 12 weeks post-irradiation, the mice were randomly grouped into three categories and irradiated with 0, 0.5 or 1.0 Gy for each group. At 2 weeks post-irradiation, mice were sacrificed to evaluate early effects of irradiation. White blood cell (WBC) counts in the PB (left panel) and chimerism of CD45+ human (middle panel) and mouse (right panel) hematopoietic cells in the PB are presented as scatter diagrams with median values (bars) (n = 9 each). PB data of NOG mice just before the irradiation (Pre) are presented as controls (n = 27). (B) The percentages of CD11b+ myeloid cells, CD19+ B cells, and CD3+ T cells in CD45+ human hematopoietic cells in the PB of NOG mice just before the irradiation (Pre, n = 27) and at 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (C) Total BM cell numbers (left panel) and chimerism of CD45+ human (middle) and mouse (right) hematopoietic cells in the BM just before the irradiation (Pre, n = 6) and at 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (D) Absolute numbers of CD34+CD38- HSCs, CD34+CD38+ HPCs, CMPs and LMPPs in the BM (bilateral femurs and tibiae) just before the irradiation (Pre, n = 6) and 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (E) Numbers of colony-forming cells included in 1,000 CD34+ human HSPCs recovered at 2 weeks after irradiation (0–1.0 Gy, n = 3 each). Bars in scatter diagrams indicate median values. P-values for trend were calculated by nonparametric Jonckheere-Terpstra test.

Mentions: At 2 weeks post-irradiation, mice were sacrificed to evaluate early effects of radiation (9 mice each). In contrast to NOD/Rag2 IL2rγ mice, hematopoietic cells of NOG mice are susceptible to irradiation due to the scid/scid background [23]. As expected, chimerism of mouse CD45+ hematopoietic cells declined in a radiation dose-dependent manner in the PB while that of human hematopoietic cells increased (Fig 2A). Among human cells in the PB, chimerism of B cells significantly decreased in a radiation dose-dependent manner while that of myeloid and T cells increased (Fig 2B). Of interest, chimerism of human CD45+ hematopoietic cells declined in a radiation dose-dependent manner in the BM while that of mouse hematopoietic cells increased probably because of inappropriate support of human HSPCs by mouse BM niche cells (Fig 2C). As a consequence, human cells were gradually outcompeted by mouse cells in the PB afterwards as we have reported previously (Figs 3 and 4) [19]. In the BM, radiation dose-dependent decreases in both proportion (data not shown) and absolute numbers were commonly observed in human CD34+CD38- HSCs, CD34+CD38+ HPCs, CD34+CD38+CD45RA-CD10-CD7-Lineage marker- common myeloid progenitors (CMPs) and CD34+CD38+CD45RA+CD62LhiLineage marker- lymphoid-primed multipotent progenitors (LMPPs) (Fig 2D). In addition, CD34+ HSPCs recovered from recipient mice at 2 weeks post-irradiation gave rise to fewer colonies compared with non-irradiated HSPCs (0 Gy vs. 0.5 Gy, P = 0.015 and 0 Gy vs. 1.0 Gy P = 0.003 by Dunnett t-test) (Fig 2E).


Non-Lethal Ionizing Radiation Promotes Aging-Like Phenotypic Changes of Human Hematopoietic Stem and Progenitor Cells in Humanized Mice.

Wang C, Oshima M, Sashida G, Tomioka T, Hasegawa N, Mochizuki-Kashio M, Nakajima-Takagi Y, Kusunoki Y, Kyoizumi S, Imai K, Nakachi K, Iwama A - PLoS ONE (2015)

Early effects of irradiation on human HSPCs in humanized mice.(A) Changes in chimerism of human hematopoietic cells in the PB after irradiation. At 12 weeks post-irradiation, the mice were randomly grouped into three categories and irradiated with 0, 0.5 or 1.0 Gy for each group. At 2 weeks post-irradiation, mice were sacrificed to evaluate early effects of irradiation. White blood cell (WBC) counts in the PB (left panel) and chimerism of CD45+ human (middle panel) and mouse (right panel) hematopoietic cells in the PB are presented as scatter diagrams with median values (bars) (n = 9 each). PB data of NOG mice just before the irradiation (Pre) are presented as controls (n = 27). (B) The percentages of CD11b+ myeloid cells, CD19+ B cells, and CD3+ T cells in CD45+ human hematopoietic cells in the PB of NOG mice just before the irradiation (Pre, n = 27) and at 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (C) Total BM cell numbers (left panel) and chimerism of CD45+ human (middle) and mouse (right) hematopoietic cells in the BM just before the irradiation (Pre, n = 6) and at 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (D) Absolute numbers of CD34+CD38- HSCs, CD34+CD38+ HPCs, CMPs and LMPPs in the BM (bilateral femurs and tibiae) just before the irradiation (Pre, n = 6) and 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (E) Numbers of colony-forming cells included in 1,000 CD34+ human HSPCs recovered at 2 weeks after irradiation (0–1.0 Gy, n = 3 each). Bars in scatter diagrams indicate median values. P-values for trend were calculated by nonparametric Jonckheere-Terpstra test.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4498777&req=5

pone.0132041.g002: Early effects of irradiation on human HSPCs in humanized mice.(A) Changes in chimerism of human hematopoietic cells in the PB after irradiation. At 12 weeks post-irradiation, the mice were randomly grouped into three categories and irradiated with 0, 0.5 or 1.0 Gy for each group. At 2 weeks post-irradiation, mice were sacrificed to evaluate early effects of irradiation. White blood cell (WBC) counts in the PB (left panel) and chimerism of CD45+ human (middle panel) and mouse (right panel) hematopoietic cells in the PB are presented as scatter diagrams with median values (bars) (n = 9 each). PB data of NOG mice just before the irradiation (Pre) are presented as controls (n = 27). (B) The percentages of CD11b+ myeloid cells, CD19+ B cells, and CD3+ T cells in CD45+ human hematopoietic cells in the PB of NOG mice just before the irradiation (Pre, n = 27) and at 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (C) Total BM cell numbers (left panel) and chimerism of CD45+ human (middle) and mouse (right) hematopoietic cells in the BM just before the irradiation (Pre, n = 6) and at 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (D) Absolute numbers of CD34+CD38- HSCs, CD34+CD38+ HPCs, CMPs and LMPPs in the BM (bilateral femurs and tibiae) just before the irradiation (Pre, n = 6) and 2 weeks after irradiation (0–1.0 Gy, n = 9 each). (E) Numbers of colony-forming cells included in 1,000 CD34+ human HSPCs recovered at 2 weeks after irradiation (0–1.0 Gy, n = 3 each). Bars in scatter diagrams indicate median values. P-values for trend were calculated by nonparametric Jonckheere-Terpstra test.
Mentions: At 2 weeks post-irradiation, mice were sacrificed to evaluate early effects of radiation (9 mice each). In contrast to NOD/Rag2 IL2rγ mice, hematopoietic cells of NOG mice are susceptible to irradiation due to the scid/scid background [23]. As expected, chimerism of mouse CD45+ hematopoietic cells declined in a radiation dose-dependent manner in the PB while that of human hematopoietic cells increased (Fig 2A). Among human cells in the PB, chimerism of B cells significantly decreased in a radiation dose-dependent manner while that of myeloid and T cells increased (Fig 2B). Of interest, chimerism of human CD45+ hematopoietic cells declined in a radiation dose-dependent manner in the BM while that of mouse hematopoietic cells increased probably because of inappropriate support of human HSPCs by mouse BM niche cells (Fig 2C). As a consequence, human cells were gradually outcompeted by mouse cells in the PB afterwards as we have reported previously (Figs 3 and 4) [19]. In the BM, radiation dose-dependent decreases in both proportion (data not shown) and absolute numbers were commonly observed in human CD34+CD38- HSCs, CD34+CD38+ HPCs, CD34+CD38+CD45RA-CD10-CD7-Lineage marker- common myeloid progenitors (CMPs) and CD34+CD38+CD45RA+CD62LhiLineage marker- lymphoid-primed multipotent progenitors (LMPPs) (Fig 2D). In addition, CD34+ HSPCs recovered from recipient mice at 2 weeks post-irradiation gave rise to fewer colonies compared with non-irradiated HSPCs (0 Gy vs. 0.5 Gy, P = 0.015 and 0 Gy vs. 1.0 Gy P = 0.003 by Dunnett t-test) (Fig 2E).

Bottom Line: Of interest, non-lethal radiation compromised contribution of HSPCs to the peripheral blood cells, particularly to CD19+ B lymphocytes, which resulted in myeloid-biased repopulation.Co-culture of limiting numbers of CD34+ HSPCs with stromal cells revealed that the frequency of B cell-producing CD34+ HSPCs at 2 weeks post-irradiation was reduced more than 10-fold.Given that compromised repopulating capacity and myeloid-biased differentiation are representative phenotypes of aged HSCs, our findings indicate that non-lethal ionizing radiation is one of the critical external stresses that promote aging of human HSPCs in the bone marrow niche.

View Article: PubMed Central - PubMed

Affiliation: Department of Cellular and Molecular Medicine, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan; College of Life Sciences, Inner Mongolia University, Hohhot 010021, China.

ABSTRACT
Precise understanding of radiation effects is critical to develop new modalities for the prevention and treatment of radiation-induced damage. We previously reported that non-lethal doses of X-ray irradiation induce DNA damage in human hematopoietic stem and progenitor cells (HSPCs) reconstituted in NOD/Shi-scid IL2rγ (NOG) immunodeficient mice and severely compromise their repopulating capacity. In this study, we analyzed in detail the functional changes in human HSPCs in NOG mice following non-lethal radiation. We transplanted cord blood CD34+ HSPCs into NOG mice. At 12 weeks post-transplantation, the recipients were irradiated with 0, 0.5, or 1.0 Gy. At 2 weeks post-irradiation, human CD34+ HSPCs recovered from the primary recipient mice were transplanted into secondary recipients. CD34+ HSPCs from irradiated mice showed severely impaired reconstitution capacity in the secondary recipient mice. Of interest, non-lethal radiation compromised contribution of HSPCs to the peripheral blood cells, particularly to CD19+ B lymphocytes, which resulted in myeloid-biased repopulation. Co-culture of limiting numbers of CD34+ HSPCs with stromal cells revealed that the frequency of B cell-producing CD34+ HSPCs at 2 weeks post-irradiation was reduced more than 10-fold. Furthermore, the key B-cell regulator genes such as IL-7R and EBF1 were downregulated in HSPCs upon 0.5 Gy irradiation. Given that compromised repopulating capacity and myeloid-biased differentiation are representative phenotypes of aged HSCs, our findings indicate that non-lethal ionizing radiation is one of the critical external stresses that promote aging of human HSPCs in the bone marrow niche.

No MeSH data available.


Related in: MedlinePlus