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Yoriko Saito,
Hitomi Yuki,
Mitsuo Kuratani,
Yoshinobu Hashizume,
Shinsuke Takagi,
Teruki Honma,
Akiko Tanaka,
Mikako Shirouzu,
Junko Mikuni,
Noriko Handa, [......],
Nahoko Suzuki,
Hiroshi Kajita,
Yuki Aoki,
Osamu Ohara,
Leonard D Shultz,
Takehiro Fukami,
Toshio Goto,
Shuichi Taniguchi,
Shigeyuki Yokoyama, Fumihiko Ishikawa
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ABSTRACT: Leukemia stem cells (LSCs) that survive conventional chemotherapy are thought to contribute to disease relapse, leading to poor long-term outcomes for patients with acute myeloid leukemia (AML). We previously identified a Src-family kinase (SFK) member, hematopoietic cell kinase (HCK), as a molecular target that is highly differentially expressed in human primary LSCs compared with human normal hematopoietic stem cells (HSCs). We performed a large-scale chemical library screen that integrated a high-throughput enzyme inhibition assay, in silico binding prediction, and crystal structure determination and found a candidate HCK inhibitor, RK-20449, a pyrrolo-pyrimidine derivative with an enzymatic IC50 (half maximal inhibitory concentration) in the subnanomolar range. A crystal structure revealed that RK-20449 bound the activation pocket of HCK. In vivo administration of RK-20449 to nonobese diabetic (NOD)/severe combined immunodeficient (SCID)/IL2rgnull mice engrafted with highly aggressive therapy-resistant AML significantly reduced human LSC and non-stem AML burden. By eliminating chemotherapy-resistant LSCs, RK-20449 may help to prevent relapse and lead to improved patient outcomes in AML.
Science translational medicine 01/2013; 5(181):181ra52. · 7.80 Impact Factor
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Mitsuhiro Fukata, Fumihiko Ishikawa,
Yuho Najima,
Takuji Yamauchi,
Yoriko Saito,
Katsuto Takenaka,
Kohta Miyawaki,
Hideki Shimazu,
Kazuya Shimoda,
Takaaki Kanemaru,
Kei-Ichiro Nakamura,
Keita Odashiro,
Koji Nagafuji,
Mine Harada,
Koichi Akashi
[show abstract]
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ABSTRACT: BACKGROUND: Definite identification of the cell types and the mechanism relevant to cardiomyogenesis is essential for effective cardiac regenerative medicine. We aimed to identify the cell populations that can generate cardiomyocytes and to clarify whether generation of donor-marker(+) cardiomyocytes requires cell fusion between BM-derived cells and recipient cardiomyocytes. METHODOLOGYPRINCIPAL FINDINGS: Purified BM stem/progenitor cells from green fluorescence protein (GFP) mice were transplanted into C57BL/6 mice or cyan fluorescence protein (CFP)-transgenic mice. Purified human hematopoietic stem cells (HSCs) from cord blood were transplanted into immune-compromised NOD/SCID/IL2rγ(null) mice. GFP(+) cells in the cardiac tissue were analyzed for the antigenecity of a cardiomyocyte by confocal microscopy following immunofluorescence staining. GFP(+) donor-derived cells, GFP(+)CFP(+) fused cells, and CFP(+) recipient-derived cells were distinguished by linear unmixing analysis. Hearts of xenogeneic recipients were evaluated for the expression of human cardiomyocyte genes by real-time quantitative polymerase chain reaction. In C57BL/6 recipients, Lin(-/low)CD45(+) hematopoietic cells generated greater number of GFP(+) cardiomyocytes than Lin(-/low)CD45(-) mesenchymal cells (37.0+/-23.9 vs 0.00+/-0.00 GFP(+) cardiomyocytes per a recipient, P = 0.0095). The number of transplanted purified HSCs (Lin(-/low)Sca-1(+) or Lin(-)Sca-1(+)c-Kit(+) or CD34(-)Lin(-)Sca-1(+)c-Kit(+)) showed correlation to the number of GFP(+) cardiomyocytes (P<0.05 in each cell fraction), and the incidence of GFP(+) cardiomyocytes per injected cell dose was greatest in CD34(-)Lin(-)Sca-1(+)c-Kit(+) recipients. Of the hematopoietic progenitors, total myeloid progenitors generated greater number of GFP(+) cardiomyocytes than common lymphoid progenitors (12.8+/-10.7 vs 0.67+/-1.00 GFP(+) cardiomyocytes per a recipient, P = 0.0021). In CFP recipients, all GFP(+) cardiomyocytes examined coexpressed CFP. Human troponin C and myosin heavy chain 6 transcripts were detected in the cardiac tissue of some of the xenogeneic recipients. CONCLUSIONSSIGNIFICANCE: Our results indicate that HSCs resulted in the generation of cardiomyocytes via myeloid intermediates by fusion-dependent mechanism. The use of myeloid derivatives as donor cells could potentially allow more effective cell-based therapy for cardiac repair.
PLoS ONE 01/2013; 8(5):e62506. · 4.09 Impact Factor
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Shunichi Suzuki,
Masaki Iwamoto,
Yoriko Saito,
Daiichiro Fuchimoto,
Shoichiro Sembon,
Misae Suzuki,
Satoshi Mikawa,
Michiko Hashimoto,
Yuki Aoki,
Yuho Najima, [......],
Nahoko Suzuki,
Emi Suzuki,
Masanori Kubo,
Jun Mimuro,
Yuji Kashiwakura,
Seiji Madoiwa,
Yoichi Sakata,
Anthony C F Perry, Fumihiko Ishikawa,
Akira Onishi
[show abstract]
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ABSTRACT: A porcine model of severe combined immunodeficiency (SCID) promises to facilitate human cancer studies, the humanization of tissue for xenotransplantation, and the evaluation of stem cells for clinical therapy, but SCID pigs have not been described. We report here the generation and preliminary evaluation of a porcine SCID model. Fibroblasts containing a targeted disruption of the X-linked interleukin-2 receptor gamma chain gene, Il2rg, were used as donors to generate cloned pigs by serial nuclear transfer. Germline transmission of the Il2rg deletion produced healthy Il2rg(+/-) females, while Il2rg(-/Y) males were athymic and exhibited markedly impaired immunoglobulin and T and NK cell production, robustly recapitulating human SCID. Following allogeneic bone marrow transplantation, donor cells stably integrated in Il2rg(-/Y) heterozygotes and reconstituted the Il2rg(-/Y) lymphoid lineage. The SCID pigs described here represent a step toward the comprehensive evaluation of preclinical cellular regenerative strategies.
Cell stem cell 06/2012; 10(6):753-8. · 23.56 Impact Factor
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ABSTRACT: Although physiological development of human lymphoid subsets has become well documented in humanized mice, in vivo development of human myeloid subsets in a xenotransplantation setting has remained unevaluated. Therefore, we investigated in vivo differentiation and function of human myeloid subsets in NOD/SCID/IL2rγ(null) (NSG) mouse recipients transplanted with purified lineage(-)CD34(+)CD38(-) cord blood hematopoietic stem cells. At 4-6 mo posttransplantation, we identified the development of human neutrophils, basophils, mast cells, monocytes, and conventional and plasmacytoid dendritic cells in the recipient hematopoietic organs. The tissue distribution and morphology of these human myeloid cells were similar to those identified in humans. After cytokine stimulation in vitro, phosphorylation of STAT molecules was observed in neutrophils and monocytes. In vivo administration of human G-CSF resulted in the recruitment of human myeloid cells into the recipient circulation. Flow cytometry and confocal imaging demonstrated that human bone marrow monocytes and alveolar macrophages in the recipients displayed intact phagocytic function. Human bone marrow-derived monocytes/macrophages were further confirmed to exhibit phagocytosis and killing of Salmonella typhimurium upon IFN-γ stimulation. These findings demonstrate the development of mature and functionally intact human myeloid subsets in vivo in the NSG recipients. In vivo human myelopoiesis established in the NSG humanized mouse system may facilitate the investigation of human myeloid cell biology including in vivo analyses of infectious diseases and therapeutic interventions.
The Journal of Immunology 05/2012; 188(12):6145-55. · 5.79 Impact Factor
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Shinsuke Takagi,
Yoriko Saito,
Atsushi Hijikata,
Satoshi Tanaka,
Takashi Watanabe,
Takanori Hasegawa,
Shinobu Mochizuki,
Jun Kunisawa,
Hiroshi Kiyono,
Haruhiko Koseki,
Osamu Ohara,
Takashi Saito,
Shuichi Taniguchi,
Leonard D Shultz, Fumihiko Ishikawa
[show abstract]
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ABSTRACT: In recent years, advances in the humanized mouse system have led to significantly increased levels of human hematopoietic stem cell (HSC) engraftment. The remaining limitations in human HSC engraftment and function include lymphoid-skewed differentiation and inefficient myeloid development in the recipients. Limited human HSC function may partially be attributed to the inability of the host mouse microenvironment to provide sufficient support to human hematopoiesis. To address this problem, we created membrane-bound human stem cell factor (SCF)/KIT ligand (KL)-expressing NOD/SCID/IL2rgKO (hSCF Tg NSG) mice. hSCF Tg NSG recipients of human HSCs showed higher levels of both human CD45(+) cell engraftment and human CD45(+)CD33(+) myeloid development compared with NSG recipients. Expression of hSCF/hKL accelerated the differentiation of the human granulocyte lineage cells in the recipient bone marrow. Human mast cells were identified in bone marrow, spleen, and gastrointestinal tissues of the hSCF Tg NSG recipients. This novel in vivo humanized mouse model demonstrates the essential role of membrane-bound hSCF in human myeloid development. Moreover, the hSCF Tg NSG humanized recipients may facilitate investigation of in vivo differentiation, migration, function, and pathology of human mast cells.
Blood 01/2012; 119(12):2768-77. · 9.90 Impact Factor
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Yoshikane Kikushige, Fumihiko Ishikawa,
Toshihiro Miyamoto,
Takahiro Shima,
Shingo Urata,
Goichi Yoshimoto,
Yasuo Mori,
Tadafumi Iino,
Takuji Yamauchi,
Tetsuya Eto,
Hiroaki Niiro,
Hiromi Iwasaki,
Katsuto Takenaka,
Koichi Akashi
[show abstract]
[hide abstract]
ABSTRACT: We report here that in chronic lymphocytic leukemia (CLL), the propensity to generate clonal B cells has been acquired already at the hematopoietic stem cell (HSC) stage. HSCs purified from patients with CLL displayed lymphoid-lineage gene priming and produced a high number of polyclonal B cell progenitors. Strikingly, their maturation into B cells was restricted always to mono- or oligo-clones with CLL-like phenotype in xenogeneic recipients. These B cell clones were independent of the original CLL clones because they had their own immunoglobulin VDJ genes. Furthermore, they used preferentially VH genes frequently used in human CLL, presumably reflecting the role of B cell receptor signaling in clonal selection. These data suggest that HSCs can be involved in leukemogenesis even in mature lymphoid tumors.
Cancer cell 08/2011; 20(2):246-59. · 25.29 Impact Factor
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ABSTRACT: Adult T-cell leukemia/lymphoma (ATLL) is a T-cell malignancy associated with HTLV-1. The HTLV-1 provirus genome has the pX region that encodes tax and HTLV-1 basic leucine zipper factor (HBZ). Previous studies have reported that the tax gene is expressed in few ATLL cases, but the HBZ gene in all ATLL cases. In this study, we used HBZ gene in situ hybridization (HBZ-ISH) for detection of the HBZ gene in formalin-fixed paraffin-embedded tissues. This method showed that all cases (n = 19) were positive for the ATLL cell line (MT-1, MT-2, and MT-4) and ATLL mouse model (HBZ-Tg mice and NOD/SCID/β2-microglobulin(null) mice with ATLL transplanted), and the HBZ gene was also detected in all human ATLL cases (n = 16). The percentage of positive cells in HBZ-ISH was 5-70%. Immunohistochemical staining for Tax protein showed positivity in seven of 11 cases in NOD/SCID/β2-microglobulin(null) mice with ATLL transplanted and in six of eight human ATLL cases, but the percentage of positive cells was very low (range, 1-5%). Although HBZ-ISH is unsuitable to detect HTLV-1 clonality, this method is convenient and can be useful for the histological diagnosis of ATLL in HTLV-1 sero-indeterminate patients.
Cancer Science 03/2011; 102(7):1432-6. · 3.33 Impact Factor
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Jun An,
Hiroshi Fujiwara,
Koichiro Suemori,
Toshiyuki Niiya,
Taichi Azuma,
Kazushi Tanimoto,
Toshiki Ochi,
Yoshiki Akatsuka,
Junichi Mineno,
Hidetoshi Ozawa, Fumihiko Ishikawa,
Kiyotaka Kuzushima,
Masaki Yasukawa
[show abstract]
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ABSTRACT: Peripheral T-cell lymphoma (PTCL) is a biologically diverse lymphoid malignancy. The clinical aggressiveness associated with hemophagocytic syndrome (HS) is a characteristic of PTCL, being more distinctive in CD8(+) PTCL. However, the underlying mechanism of PTCL-associated HS has not yet been fully investigated. We newly established a novel IL-2-dependent CD8(+) PTCL lymphoma cell line (T8ML-1) from a patient with CD8(+) PTCL who suffered recurrent HS accompanying disease flare-up. Focusing on the lymphoma cell T-cell receptor (TCR), we examined the lymphoma cell functions responsible for such clinical manifestations. First, T8ML-1.1 in which endogenous TCR-α/β chains were silenced by siRNAs, and T8ML-1.2 in which endogenous TCR-α/β chains were replaced with HLA-A*24:02-restricted and WT1(235-243)-specific TCR-α/β, were established. T8ML-1 exerted phytohemagglutinin (PHA)-dependent cytotoxicity via granular exocytosis. Additionally, soluble factors produced by PHA-stimulated T8ML-1, which included INF-γ and TNF-α, but not by simple-cultured T8ML-1, caused human monocytes to exhibit erythrophagocytosis and thrombophagocytosis in vitro. PHA binding induced phosphorylation of CD3ζ chain. Furthermore, both cytotoxicity and hemophagocytosis were completely inhibited by T8ML-1.1, but eventually restored by T8ML-1.2. These data suggest that exogenous activation of TCR signaling in PTCL cells might play an important role in the formation of PTCL-associated HS.
International journal of hematology 02/2011; 93(2):176-85. · 1.17 Impact Factor
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Noriaki Kawano,
Shuro Yoshida,
Nobuyuki Ono,
Daisuke Himeji,
Yuri Nagahiro,
Sayaka Kawano,
Kiyoshi Yamashita,
Naoko Ikeda,
Shigehiro Uezono,
Hidenobu Ochiai,
Fumiko Kawano,
Ikuo Kikuchi, Fumihiko Ishikawa,
Kazuya Shimoda,
Akira Ueda,
Koichi Akashi
[show abstract]
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ABSTRACT: Disseminated intravascular coagulation (DIC) is a clinical entity with high mortality and is characterized by multiple organ failure caused by activation of systemic intravascular coagulation. Although a standard treatment for DIC has not been established owing to the absence of randomized controlled trials, recent reports have indicated that recombinant human soluble thrombomodulin (rTM) is effective against DIC. To elucidate the clinical characteristics and outcomes of DIC, we retrospectively analyzed 35 DIC patients treated with rTM at our institution over a 2-year period (infectious disease: 21 cases; hematological disease: 14 cases). Diagnosis of DIC was based on the diagnostic criteria for DIC of the Japanese Ministry of Health and Welfare. In addition to the treatment of underlying diseases, we administered rTM for 6 consecutive days. Twenty-one (60.0%) of the DIC patients attained resolution of DIC at 7 days after administration (infectious disease: 61.9%; hematological disease: 57.1%). Furthermore, 7 of the remaining 14 DIC patients (who did not attain resolution at 7 days) attained resolution at an average of 12.1 days. Consequently, 28 (80.0%) of the 35 patients were alive with resolution of DIC after a 28-day observation period (infectious disease: 76.2%; hematological disease: 85.7%). Among them, for 7 (70%) of the 10 DIC patients with severe life-threatening bleeding symptoms without hemorrhagic shock, treatment with heparin was contraindicated; these patients were successfully treated with rTM without the progression of hemorrhage. In the majority of DIC patients, rTM administration may be an effective, safe, and feasible therapeutic modality producing a good outcome.
Journal of Clinical and Experimental Hematopathology 01/2011; 51(2):101-7.
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Leonard D Shultz,
Yoriko Saito,
Yuho Najima,
Satoshi Tanaka,
Toshiki Ochi,
Mariko Tomizawa,
Takehiko Doi,
Akiko Sone,
Nahoko Suzuki,
Hiroshi Fujiwara,
Masaki Yasukawa, Fumihiko Ishikawa
[show abstract]
[hide abstract]
ABSTRACT: Whereas humanized mouse models have contributed significantly to human immunology research, human T cells developing in mouse thymic environment fail to demonstrate HLA-restricted function. To achieve HLA-restricted human immune response, we created an immune-compromised non-obese diabetic/SCID/IL2rg(null) strain (NSG) with homozygous expression of HLA class I heavy chain and light chain (NSG-HLA-A2/HHD). Transplantation of purified Lin-CD34+CD38- human hematopoietic stem cells into NSG-HLA-A2/HHD newborns resulted in the development of human CD4+ and CD8+ TCR alphabeta+ T cells and CD4-CD8- and CD8+ TCR gammadelta+ cells in recipient bone marrow and spleen. Human cytotoxic T lymphocytes (CTLs) become functionally mature, as evidenced by the production of granzyme corresponding to phenotypic transition from naïve to effector memory CTLs. In these recipients, human Th17 cells developed along with Th1 and Th2 cells. Epstein-Barr virus (EBV) infection in the humanized NSG-HLA-A2/HHD recipients resulted in the formation of lymphoproliferative lesions consisting mainly of human B cells with scattered human T cells. Human CTLs developing in the recipients recognized EBV-derived peptides in an HLA-restricted manner and exerted HLA-restricted cytotoxicity against EBV-infected human B cells. The HLA-expressing humanized mouse with functional HLA-restricted T cells and consistent representation of rare T-cell subsets overcomes a major constraint in human immunology, and serves as a useful model for investigation of human immune responses against pathogens and for the development of therapeutic strategies against human diseases.
Proceedings of the National Academy of Sciences 07/2010; 107(29):13022-7. · 9.68 Impact Factor
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Yoriko Saito,
Naoyuki Uchida,
Satoshi Tanaka,
Nahoko Suzuki,
Mariko Tomizawa-Murasawa,
Akiko Sone,
Yuho Najima,
Shinsuke Takagi,
Yuki Aoki,
Atsushi Wake,
Shuichi Taniguchi,
Leonard D Shultz, Fumihiko Ishikawa
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ABSTRACT: Cancer stem cells have been proposed to be important for initiation, maintenance and recurrence of various malignancies, including acute myeloid leukemia (AML). We have previously reported that CD34+CD38- human primary AML stem cells residing in the endosteal region of the bone marrow are relatively chemotherapy resistant. Using a NOD/SCID/IL2rgamma(null) mouse model of human AML, we now show that the AML stem cells in the endosteal region are cell cycle quiescent and that these stem cells can be induced to enter the cell cycle by treatment with granulocyte colony-stimulating factor (G-CSF). In combination with cell cycle-dependent chemotherapy, G-CSF treatment significantly enhances induction of apoptosis and elimination of human primary AML stem cells in vivo. The combination therapy leads to significantly increased survival of secondary recipients after transplantation of leukemia cells compared with chemotherapy alone.
Nature Biotechnology 02/2010; 28(3):275-80. · 29.50 Impact Factor
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Yoriko Saito,
Hiroshi Kitamura,
Atsushi Hijikata,
Mariko Tomizawa-Murasawa,
Satoshi Tanaka,
Shinsuke Takagi,
Naoyuki Uchida,
Nahoko Suzuki,
Akiko Sone,
Yuho Najima,
Hidetoshi Ozawa,
Atsushi Wake,
Shuichi Taniguchi,
Leonard D Shultz,
Osamu Ohara, Fumihiko Ishikawa
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[hide abstract]
ABSTRACT: Human acute myeloid leukemia (AML) originates from rare leukemia stem cells (LSCs). Because these chemotherapy-resistant LSCs are thought to underlie disease relapse, effective therapeutic strategies specifically targeting these cells may be beneficial. Here, we report identification of a primary human LSC gene signature and functional characterization of human LSC-specific molecules in vivo in a mouse xenotransplantation model. In 32 of 61 (53%) patients with AML, either CD32 or CD25 or both were highly expressed in LSCs. CD32- or CD25-positive LSCs could initiate AML and were cell cycle-quiescent and chemotherapy-resistant in vivo. Normal human hematopoietic stem cells depleted of CD32- and CD25-positive cells maintained long-term multilineage hematopoietic reconstitution capacity in vivo, indicating the potential safety of treatments targeting these molecules. In addition to CD32 and CD25, quiescent LSCs within the bone marrow niche also expressed the transcription factor WT1 and the kinase HCK. These molecules are also promising targets for LSC-specific therapy.
Science translational medicine 02/2010; 2(17):17ra9. · 7.80 Impact Factor
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Tetsuya Takayama,
Teruyoshi Kondo,
Masatoshi Kobayashi,
Keisuke Ohta,
Yoshihiro Ishibashi,
Takaaki Kanemaru,
Hideki Shimazu, Fumihiko Ishikawa,
Takahiro Nakamura,
Shigeru Kinoshita,
Kei-Ichiro Nakamura
[show abstract]
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ABSTRACT: Enhanced green fluorescence protein (eGFP)-labeled bone marrow (BM) cells were transplanted into syngeneic C57BL/6 (wild-type) mice to investigate the distribution pattern, immunohistochemical characteristics, three-dimensional structure, and ultrastructure of the BM-derived cells in the mouse cornea using a fluorescence microscope, a confocal laser scanning microscope, and a transmission electron microscope. This study provided direct evidence that two morphologically distinct types of BM-derived cells were distributed in the mouse cornea. The majority of the GFP+ cells showed a flattened polygonal form with obtuse angles and these cells were distributed in the corneal stroma. The other type was the GFP+ cells demonstrating slim cell bodies with long and extremely thin dendrites and which were distributed in the corneal epithelium. The immunohistochemical characteristics and ultrastructure of BM-derived cells suggest that most of these cells have a macrophage lineage, whereas some cells in the corneal stroma do not. Interestingly, the direct intimate contact between GFP-labeled BM derived cells and non-GFP-labeled resident cells within the corneal stroma were also clearly visualized at the fine structural level. These data provide new and more detailed insight into the nature of BM-derived cells in the cornea.
The Anatomical Record Advances in Integrative Anatomy and Evolutionary Biology 03/2009; 292(5):756-63. · 1.47 Impact Factor
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12/2008: pages 121-132;
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ABSTRACT: Ex vivo hematopoiesis from embryonic sources offers exciting promises in basic research and medicine. In this issue of Cell Stem Cell, Ledran et al. (2008) describe human embryonic stem cell (hESC)-derived hematopoiesis, while Taoudi et al. (2008) define the origin of definitive hematopoietic stem cells (HSCs) from the mouse aorta-gonad-mesonephros (AGM) region.
Cell stem cell 08/2008; 3(1):8-10. · 23.56 Impact Factor
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Yoshikane Kikushige,
Goichi Yoshimoto,
Toshihiro Miyamoto,
Tadafumi Iino,
Yasuo Mori,
Hiromi Iwasaki,
Hiroaki Niiro,
Katsuto Takenaka,
Koji Nagafuji,
Mine Harada, Fumihiko Ishikawa,
Koichi Akashi
[show abstract]
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ABSTRACT: FLT3/FLK2, a member of the receptor tyrosine kinase family, plays a critical role in maintenance of hematopoietic homeostasis, and the constitutively active form of the FLT3 mutation is one of the most common genetic abnormalities in acute myelogenous leukemia. In murine hematopoiesis, Flt3 is not expressed in self-renewing hematopoietic stem cells, but its expression is restricted to the multipotent and the lymphoid progenitor stages at which cells are incapable of self-renewal. We extensively analyzed the expression of Flt3 in human (h) hematopoiesis. Strikingly, in both the bone marrow and the cord blood, the human hematopoietic stem cell population capable of long-term reconstitution in xenogeneic hosts uniformly expressed Flt3. Furthermore, human Flt3 is expressed not only in early lymphoid progenitors, but also in progenitors continuously along the granulocyte/macrophage pathway, including the common myeloid progenitor and the granulocyte/macrophage progenitor. We further found that human Flt3 signaling prevents stem and progenitors from spontaneous apoptotic cell death at least through up-regulating Mcl-1, an indispensable survival factor for hematopoiesis. Thus, the distribution of Flt3 expression is considerably different in human and mouse hematopoiesis, and human FLT3 signaling might play an important role in cell survival, especially at stem and progenitor cells that are critical cellular targets for acute myelogenous leukemia transformation.
The Journal of Immunology 07/2008; 180(11):7358-67. · 5.79 Impact Factor
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ABSTRACT: Pancreatic acinar cells and hepatocytes arise from the same cell population located within the embryonic endoderm. It has been reported that a multipotent population of liver cells is capable of differentiating into pancreatic cells. Recent studies revealed that murine and human hematopoietic cells could generate hepatocytes in vivo. Based on this developmental proximity between liver and pancreatic acinar cells, we examined whether human cord blood (CB) cells can generate pancreatic cells in vivo using a murine xenograft model.
We transplanted 1 x 10 CD34 human CB cells into "conditioned" newborn nonobese diabetic-severe combined immunodeficiency/beta-2 microglobulin-null mice via facial vein injection and, 3 to 4 months later, examined the pancreata from recipient mice showing high-level human multilineage hematopoietic engraftment in the bone marrow.
Reverse transcriptase-polymerase chain reaction and immunohistochemical analyses revealed human amylase mRNA and protein expression, respectively, in the pancreata from recipient mice. Using fluorescence in situ hybridization, we identified human alpha-satellite, DNA-positive cells with a morphology characteristic of pancreatic acinar cells. We also identified cells in paraffin sections of the pancreata that expressed amylase mRNA, had morphological characteristics of acinar cells, and contained human but not mouse centromeric DNA.
These findings establish that human umbilical CB cells are capable of generating pancreatic acinar cells via a nonfusion mechanism.
Pancreas 04/2008; 36(2):e30-5. · 2.39 Impact Factor
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Eri Miyata,
Masahiro Masuya,
Shuro Yoshida,
Shiho Nakamura,
Keizo Kato,
Yuka Sugimoto,
Tetsunori Shibasaki,
Kentaro Yamamura,
Kohshi Ohishi,
Kazuhiro Nishii, Fumihiko Ishikawa,
Hiroshi Shiku,
Naoyuki Katayama
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ABSTRACT: Hepatic stellate cells are believed to play a key role in the development of liver fibrosis. Several studies have reported that bone marrow cells can give rise to hepatic stellate cells. We hypothesized that hepatic stellate cells are derived from hematopoietic stem cells. To test this hypothesis, we generated chimeric mice by transplantation of clonal populations of cells derived from single enhanced green fluorescent protein (EGFP)-marked Lin(-)Sca-1(+)c-kit(+)CD34(-) cells and examined the histology of liver tissues obtained from the chimeric mice with carbon tetrachloride (CCl(4))-induced injury. After 12 weeks of CCl(4) treatment, we detected EGFP(+) cells in the liver, and some cells contained intracytoplasmic lipid droplets. Immunofluorescence analysis demonstrated that 50% to 60% of the EGFP(+) cells were negative for CD45 and positive for vimentin, glial fibrillary acidic protein, ADAMTS13, and alpha-smooth muscle actin. Moreover, EGFP(+) cells isolated from the liver synthesized collagen I in culture. These phenotypes were consistent with those of hepatic stellate cells. The hematopoietic stem cell-derived hepatic stellate cells seen in male-to-male transplants revealed only one Y chromosome. Our findings suggest that hematopoietic stem cells contribute to the generation of hepatic stellate cells after liver injury and that the process does not involve cell fusion.
Blood 03/2008; 111(4):2427-35. · 9.90 Impact Factor
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Fumihiko Ishikawa,
Hiroaki Niiro,
Tadafumi Iino,
Shuro Yoshida,
Noriyuki Saito,
Shinya Onohara,
Toshihiro Miyamoto,
Hiroko Minagawa,
Shin-Ichiro Fujii,
Leonard D Shultz,
Mine Harada,
Koichi Akashi
[show abstract]
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ABSTRACT: Two distinct dendritic cell (DC) subsets, conventional DCs (cDCs) and plasmacytoid DCs (pDCs), have been shown to develop via either the myeloid or the lymphoid pathway in murine hematopoiesis. Lineage-specific phenotypes or functions of "myeloid" and "lymphoid" DCs, however, still remain elusive. Furthermore, such analysis has been particularly difficult in humans, due to lack of an assay system appropriate for the analysis of human stem and progenitor cell differentiation. Here, using a highly efficient xenotransplantation model, we extensively analyze the origin and the molecular signature of human DCs. Purified human common myeloid progenitors (CMPs) and common lymphoid progenitors (CLPs) were intravenously transplanted into nonobese diabetic-severe combined immunodeficiency (NOD-scid)/IL2rgamma(null) newborn mice. CMPs and CLPs displayed significant expansion in the xenogeneic host, and human cDC and pDC progeny were isolatable. Strikingly, each human DC subset possessed indistinguishable expression patterns of surface phenotype and gene transcripts regardless of their CMP or CLP origin, even at the genome-wide level. Thus, cDC and pDC normally develop after cells have committed to the myeloid or the lymphoid lineage in human hematopoiesis, while their transcriptional signatures are well preserved irrespective of their lineage origin. We propose that human DCs use unique and flexible developmental programs that cannot be categorized into the conventional myeloid or lymphoid pathway.
Blood 12/2007; 110(10):3591-660. · 9.90 Impact Factor
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Fumihiko Ishikawa,
Shuro Yoshida,
Yoriko Saito,
Atsushi Hijikata,
Hiroshi Kitamura,
Satoshi Tanaka,
Ryu Nakamura,
Toru Tanaka,
Hiroko Tomiyama,
Noriyuki Saito,
Mitsuhiro Fukata,
Toshihiro Miyamoto,
Bonnie Lyons,
Koichi Ohshima,
Naoyuki Uchida,
Shuichi Taniguchi,
Osamu Ohara,
Koichi Akashi,
Mine Harada,
Leonard D Shultz
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ABSTRACT: Acute myelogenous leukemia (AML) is the most common adult leukemia, characterized by the clonal expansion of immature myeloblasts initiating from rare leukemic stem (LS) cells. To understand the functional properties of human LS cells, we developed a primary human AML xenotransplantation model using newborn nonobese diabetic/severe combined immunodeficient/interleukin (NOD/SCID/IL)2r gamma(null) mice carrying a complete null mutation of the cytokine gamma c upon the SCID background. Using this model, we demonstrated that LS cells exclusively recapitulate AML and retain self-renewal capacity in vivo. They home to and engraft within the osteoblast-rich area of the bone marrow, where AML cells are protected from chemotherapy-induced apoptosis. Quiescence of human LS cells may be a mechanism underlying resistance to cell cycle-dependent cytotoxic therapy. Global transcriptional profiling identified LS cell-specific transcripts that are stable through serial transplantation. These results indicate the potential utility of this AML xenograft model in the development of novel therapeutic strategies targeted at LS cells.
Nature Biotechnology 12/2007; 25(11):1315-21. · 23.27 Impact Factor
