Performance information

• Complete genome sequence of emm1 Streptococcus pyogenes 10-85, a strain isolated from a patient with streptococcal toxic shock syndrome in Japan　　　　　　　　　　　　　　　
  Ichiro Tatsuno; Masanori Isaka; Masakado Matsumoto; Naomi Nishio; Hideyuki Matsui; Tadao Hasegawa
  Microbiology Resource Announcements, Volume：8, Number：24, Jan. 2019, ［International magazine］
  Copyright © 2019 Tatsuno et al. Here, we announce the complete genome sequence of Streptococcus pyogenes strain 10-85 (type emm1), isolated from a patient with streptococcal toxic shock syndrome (STSS). The strain lacks the genomic regions encoding SalR-SalK, a two-component regulatory system, and the adjacent type I restriction modification system.
  English, Scientific journal
  DOI：https://doi.org/10.1128/MRA.00453-19
  DOI ID：10.1128/MRA.00453-19, PubMed ID：31196924, PubMed Central ID：PMC6588041, SCOPUS ID：85068464176
• Female GADD34 mice develop age-related inflammation and hepatocellular carcinoma　　　　　　　　　　　　　　　
  Naomi Nishio; Tadao Hasegawa; Ichiro Tatsuno; Masanori Isaka; Ken-ichi Isobe
  GERIATRICS & GERONTOLOGY INTERNATIONAL, Volume：17, Number：12, First page：2593, Last page：2601, Dec. 2017, [Reviewed]
  AimTo analyze the impact of sex on GADD34 function, we studied the aging of female GADD34-deficient mice and compared them with male GADD34-deficient mice.
  MethodsWe used GADD34-deficient mice on a C57BL/6 background. These mice were fed a normal diet throughout their life. Alternatively, they were fed a high-fat diet at 3months-of-age. Liver tissues taken from mice were analyzed by hematoxylin-eosin staining and immunohistochemical methods. Fresh liver cells were analyzed by flow cytometry.
  ResultsWe found that female GADD34-deficient mice did not develop obesity or fatty livers. However, female GADD34-deficient mice had infiltrations of myeloid cells in the liver, followed by liver atrophy. Many female GADD34-deficient mice developed hepatocellular carcinoma, whereas female wild-type (WT) mice did not show hepatocellular carcinoma during aging. Female GADD34-deficient mice and female WT mice developed the same percentages of lymphoma. Although a high-fat diet induced a higher level of steatosis in young male GADD34-deficient mice compared with WT mice, a high-fat diet induced the same level of steatosis in young female GADD34-deficient mice compared with WT mice. However, GADD34-deficient female young mice had a higher level of infiltration of myeloid cells and myofibroblasts than WT mice.
  ConclusionsIn contrast to male GADD34-deficient mice, female GADD34-deficient mice did not show obesity as they aged. However, similar to the males, they developed inflammation followed by hepatocellular carcinoma. Geriatr Gerontol Int 2017; 17: 2593-2601.
  WILEY, English, Scientific journal
  DOI：https://doi.org/10.1111/ggi.13080
  DOI ID：10.1111/ggi.13080, ISSN：1444-1586, eISSN：1447-0594, PubMed ID：28635009, Web of Science ID：WOS:000418347300043
• Immunological aspects of age-related diseases.　　　　　　　　　　　　　　　
  Ken-Ichi Isobe; Naomi Nishio; Tadao Hasegawa
  World journal of biological chemistry, Volume：8, Number：2, First page：129, Last page：137, May 2017, [Reviewed], ［International magazine］
  The proportion of elderly people rises in the developed countries. The increased susceptibility of the elderly to infectious diseases is caused by immune dysfunction, especially T cell functional decline. Age-related hematopoietic stem cells deviate from lymphoid lineage to myeloid lineage. Thymus shrinks early in life, which is followed by the decline of naïve T cells. T-cell receptor repertoire diversity declines by aging, which is caused by cytomegalovirus-driven T cell clonal expansion. Functional decline of B cell induces antibody affinity declines by aging. Many effector functions including phagocytosis of myeloid cells are down regulated by aging. The studies of aging of myeloid cells have some controversial results. Although M1 macrophages have been shown to be replaced by anti-inflammatory (M2) macrophages by advanced age, many human studies showed that pro-inflammatory cytokines are elevated in older human. To solve this discrepancy here we divide age-related pathological changes into two categories. One is an aging of immune cell itself. Second is involvement of immune cells to age-related pathological changes. Cellular senescence and damaged cells in aged tissue recruit pro-inflammatory M1 macrophages, which produce pro-inflammatory cytokines and proceed to age-related diseases. Underlying biochemical and metabolic studies will open nutritional treatment.
  English, Scientific journal
  DOI：https://doi.org/10.4331/wjbc.v8.i2.129
  DOI ID：10.4331/wjbc.v8.i2.129, PubMed ID：28588756, PubMed Central ID：PMC5439164
• GADD34 Promotes Tumor Growth by Inducing Myeloid-derived Suppressor Cells　　　　　　　　　　　　　　　
  Lintao Liu; Sachiko Ito; Naomi Nishio; Yang Sun; Yuriko Tanaka; Ken-Ichi Isobe
  ANTICANCER RESEARCH, Volume：36, Number：9, First page：4623, Last page：4628, Sep. 2016, [Reviewed]
  Background: Tumor hypoxia induces the expression of growth arrest and DNA damage-inducible protein (GADD34). However, the role of GADD34 in tumor growth remains unclear. Materials and Methods: Gadd34 expression was knocked-down through lentivirus-mediated short hairpin RNA (shRNA) in tumor cells, which were subsequently injected subcutaneously into mice. Tumor volumes and myeloid-derived suppressor cells (MDSCs) were monitored. Isolated MDSCs were incubated with tumor supernatant to investigate the impact of GADD34 on cytokine secretion of MDSCs. Results: We observed that reduction of GADD34 expression significantly suppressed tumor, and resulted in decreased accumulation of MDSCs and T-cells, and inhibition of GADD34 reduced secretion of vascular epithelial growth factor a and transforming growth factor beta by MDSCs. Conclusion: These findings provide a promising strategy for targeting GADD34 activity in order to inhibit tumor growth.
  INT INST ANTICANCER RESEARCH, English, Scientific journal
  DOI：https://doi.org/10.21873/anticanres.11012
  DOI ID：10.21873/anticanres.11012, ISSN：0250-7005, eISSN：1791-7530, PubMed ID：27630304, Web of Science ID：WOS:000384001800032
• Recruitment of Gr1(+)CD11b(+)F4/80(+)Population in the Bone Marrow and Spleen by Irradiation-Induced Pulmonary Damage (vol 38, pg 465, 2015)　　　　　　　　　　　　　　　
  Suganya Thanasegaran; Sachiko Ito; Naomi Nishio; Mohammad Nizam Uddin; Yang Sun; Ken-ichi Isobe
  INFLAMMATION, Volume：39, Number：1, First page：499, Last page：500, Feb. 2016, [Reviewed]
  SPRINGER/PLENUM PUBLISHERS, English
  DOI：https://doi.org/10.1007/s10753-015-0216-z
  DOI ID：10.1007/s10753-015-0216-z, ISSN：0360-3997, eISSN：1573-2576, PubMed ID：26238279, Web of Science ID：WOS:000370083500055
• Thymic Epithelial Cells Induced from Pluripotent Stem Cells by a Three-Dimensional Spheroid Culture System Regenerates Functional T Cells in Nude Mice　　　　　　　　　　　　　　　
  Motohito Okabe; Sachiko Ito; Naomi Nishio; Yuriko Tanaka; Ken-Ichi Isobe
  CELLULAR REPROGRAMMING, Volume：17, Number：5, First page：368, Last page：375, Oct. 2015, [Reviewed]
  The thymus is mainly composed of two types of epithelial cells, medullary thymic epithelial cells and cortex thymic epithelial cells (mTECs and cTECs). The tissue structure and mechanism for T cell development are complicated, with generation of the thymus regulated by complex molecular and cellular interactions of the thymic microenvironment during embryogenesis. Since the development of organ regeneration techniques became available, complete in vitro regeneration of the thymus has been attempted. Steric induction systems are thought to be optimal for tissue regeneration, but three-dimensional (3-D) induction of TECs from induced pluripotent stem cells (iPSCs) has not yet been reported. Here, we demonstrate the induction of functional TECs from iPSCs by a 3-D spheroid culture system with recruitment of robust numbers of T cells into the peripheral blood. Purified iPSC-derived TECs showed a sufficient expression level of FoxN1 comparable to TECs, and phenotypic analysis revealed that iPSC-derived TECs were expressing K5. Moreover, transplants of cell aggregations into recipient mice were not rejected and there was normal support of T cell development. Functional analysis revealed that T cells showed immune tolerance to both donor and recipient MHCs and could reject an allogeneic third party's skin graft without tumorigenesis. Taken together, these findings raised the possibility of using iPSC-derived TECs induced by 3-D spheroid culture in future regenerative therapy for patients with immunodeficiency.
  MARY ANN LIEBERT, INC, English, Scientific journal
  DOI：https://doi.org/10.1089/cell.2015.0006
  DOI ID：10.1089/cell.2015.0006, ISSN：2152-4971, eISSN：2152-4998, PubMed ID：26348437, Web of Science ID：WOS:000361523600006
• GADD34 Facilitates Cell Death Resulting from Proteasome Inhibition　　　　　　　　　　　　　　　
  Lintao Liu; Sachiko Ito; Naomi Nishio; Yang Sun; Nana Chen; Yuriko Tanaka; Ken-ichi Isobe
  ANTICANCER RESEARCH, Volume：35, Number：10, First page：5317, Last page：5324, Oct. 2015, [Reviewed]
  Background: Proteasome inhibition has been proven to be a promising therapeutic strategy in cancer clinical treatment. Inhibition of proteasome leads to failure of amino acid homeostasis, which causes cell death via activation of various mechanisms. Materials and Methods: To investigate the role of GADD34 in cell death following proteasome inhibition, we treated WT MEF and GADD34 KO MEF with MG132 and various analyses were performed, including PI staining, western blot, immunofluorescence and ROS production. Results: Expression of GADD34 dramatically enhanced MG132-induced cell death via protein synthesis. GADD34 decreased phosphorylated eIF2 alpha and increased ROS production and the levels of ubiquinated protein. Importantly, we found that accumulation of autophagy following MG132-treatment facilitated cell death in MEF. Conclusion: GADD34 plays a vital role in promoting cell death following proteasome inhibition via enhancing protein synthesis to activate death-associated mechanisms, including ER stress, ROS production and autophagy formation.
  INT INST ANTICANCER RESEARCH, English, Scientific journal
  ISSN：0250-7005, eISSN：1791-7530, PubMed ID：26408692, Web of Science ID：WOS:000361823200020
• Effects of growth arrest and DNA damage-inducible protein 34 (GADD34) on inflammation-induced colon cancer in mice　　　　　　　　　　　　　　　
  Yuriko Tanaka; Sachiko Ito; Reina Oshino; Nana Chen; Naomi Nishio; Ken-ichi Isobe
  BRITISH JOURNAL OF CANCER, Volume：113, Number：4, First page：669, Last page：679, Aug. 2015, [Reviewed]
  Background: Growth arrest and DNA damage-inducible protein 34 (GADD34/Ppp1r15a) is a family of GADD proteins that are induced by DNA damage. GADD34 protein has been suggested to regulate inflammation or host defense systems. However, the in vivo function of GADD34 in inflammation is still unclear. Long lasting inflammation, such as that seen in Crohn's disease and ulcerative colitis, is associated with a higher incidence of colorectal cancer (CRC).
  Methods: Using a colitis-associated cancer model, we analysed GADD34-deficient (KO) mice to study the effect of GADD34 on colitis and colorectal tumorigenesis.
  Results: We found a higher incidence of CRC in wild-type (WT) mice than in GADD34KO mice. Moreover, dextran sodium sulfate (DSS)-induced inflammatory responses were downregulated by GADD34 deficiency. The expression of pro-inflammatory mediators such as TNF alpha, IL-6, and iNOS/NOS2 was higher in the colons of WT mice than GADD34KO mice. IL-6 is known to activate STAT3 signalling in colonic epithelial cells and subsequently induced epithelial proliferation. We found that IL-6-STAT3 signalling and epithelial proliferation were higher in WT mice compared with GADD34KO mice.
  Conclusions: These results indicated that GADD34 upregulated pro-inflammatory mediator production leading to a higher tumour burden following azoxymethane (AOM)/DSS treatment.
  NATURE PUBLISHING GROUP, English, Scientific journal
  DOI：https://doi.org/10.1038/bjc.2015.263
  DOI ID：10.1038/bjc.2015.263, ISSN：0007-0920, eISSN：1532-1827, PubMed ID：26196182, Web of Science ID：WOS:000359438300013
• GADD34-deficient mice develop obesity, nonalcoholic fatty liver disease, hepatic carcinoma and insulin resistance　　　　　　　　　　　　　　　
  Naomi Nishio; Ken-ichi Isobe
  SCIENTIFIC REPORTS, Volume：5, First page：13519, Aug. 2015, [Reviewed]
  The prevalence of nonalcoholic fatty liver disease (NAFLD) is increasing in parallel with the prevalence of obesity. DNA damage-inducible protein 34 (GADD34/Ppp1r15a), originally isolated from UV-inducible transcripts in Chinese hamster ovary (CHO) cells, dephosphorylates several kinases that function in important signaling cascades, including dephosphorylation of eIF2 alpha. We examined the effects of GADD34 on natural life span by using GADD34-deficient mice. Here we observed for the first time that with age GADD34-deficient mice become obese, developing fatty liver followed by liver cirrhosis, hepatocellular carcinoma, and insulin resistance. We found that myofibroblasts and immune cells infiltrated the portal veins of aged GADD34-deficient mouse livers. A high-fat diet (HFD) induced a higher level of steatosis in young GADD34-deficient mice compared with WT mice. Differentiation into fat is dependent on insulin signaling. Insulin signaling in young GADD34-deficient mice was higher than that in WT mice, which explained the higher fat differentiation of mouse embryonic fibroblasts (MEFs) observed in GADD34-deficient mice. Through aging or a HFD, insulin signaling in GADD34-deficient liver converted to be down regulated compared with WT mice. We found that a HFD or palmitate treatment converted insulin signaling by up-regulating TNF-alpha and JNK.
  NATURE PUBLISHING GROUP, English, Scientific journal
  DOI：https://doi.org/10.1038/srep13519
  DOI ID：10.1038/srep13519, ISSN：2045-2322, PubMed ID：26316333, Web of Science ID：WOS:000360239500001
• Growth arrest and DNA damage-inducible protein (GADD34) enhanced liver inflammation and tumorigenesis in a diethylnitrosamine (DEN)-treated murine model　　　　　　　　　　　　　　　
  Nana Chen; Naomi Nishio; Sachiko Ito; Yuriko Tanaka; Yang Sun; Ken-ichi Isobe
  CANCER IMMUNOLOGY IMMUNOTHERAPY, Volume：64, Number：6, First page：777, Last page：789, Jun. 2015, [Reviewed]
  Growth arrest and DNA damage-inducible protein (GADD34/Ppp1r15a) is induced by various stimuli including DNA damage and ER stress. DNA damage and oncogene activation, accompanied by tumor-specific DNA repair defects and a failure to stall the cell cycle, are early markers of hepatocellular carcinoma (HCC). However, whether GADD34 accounts for regulating HCC tumorigenesis remains elusive. Here, we demonstrated that GADD34 expression was upregulated in the liver of mice after exposure to a carcinogen, diethylnitrosamine (DEN). In both acute and chronic DEN treatment models, GADD34 deficiency not only decreased oncogene expression, but also reduced hepatic damage. Moreover, loss of GADD34 attenuated immune cell infiltration, pro-inflammatory cytokine expression and hepatic compensatory proliferation. Finally, GADD34-deficient mice showed impaired hepatocarcinogenesis. Thus, the process of DEN-induced HCC proceeded as follows. First, DEN treatment induced DNA damage in hepatocytes, resulting in elevated expression of GADD34 in the liver. The increased expression of GADD34 augmented hepatic necrosis followed by elevated expression of interleukin (IL)-1 beta and monocyte chemoattractant protein 1. This process promoted immune cell infiltration and Kupffer cell/macrophage activation followed by production of reactive oxygen species and pro-tumorigenic cytokines such as IL-6 and tumor necrosis factor-alpha. The pro-tumorigenic cytokines stimulated compensatory proliferation of surviving and mutant hepatocytes. Together with oncogene c-Myc expression, these processes led to HCC. Our results suggest therapeutic opportunities for HCC by targeting GADD34-related pathways.
  SPRINGER, English, Scientific journal
  DOI：https://doi.org/10.1007/s00262-015-1690-8
  DOI ID：10.1007/s00262-015-1690-8, ISSN：0340-7004, eISSN：1432-0851, PubMed ID：25832002, Web of Science ID：WOS:000355775500014
• Recruitment of Gr1(+)CD11b(+)F4/80(+) Population in the Bone Marrow and Spleen by Irradiation-Induced Pulmonary Damage　　　　　　　　　　　　　　　
  Suganya Thanasegaran; Sachiko Ito; Naomi Nishio; Mohammad Nizam Uddin; Yang Sun; Ken-ichi Isobe
  INFLAMMATION, Volume：38, Number：2, First page：465, Last page：475, Apr. 2015, [Reviewed]
  Radiation-induced lung injury is a kind of sterile inflammation, which may lead to morbidity and mortality. The mechanism by which ionizing radiation activate the immune system is not well understood. In the present study, we have investigated the immunological responses induced by local irradiation-induced damage in mouse lung. The left lungs of C57BL/6 mice were irradiated at a high dose of 100 Gy. The histology of the lungs and spleen showed evidences of alveolar inflammation and congestion at 2 weeks after X-ray treatment. Also, prominent increase in cells expressing the cell surface markers, Gr(+)CD11b(+)F4/80(+) and Ly6C(+) Ly6G(+) were observed 2 weeks after X-ray treatment (100 Gy). Gr1(+)CD11b(+)F4/80(+) cell depletion by clodronate treatment reversed the histological effects and also failed to recruit Gr(+)CD11b(+) cells or F4/80(+) cells caused by irradiation. The origin of recruited Gr1(+)CD11b(+) cells was found to be a mixed resident and recruited phenotype.
  SPRINGER/PLENUM PUBLISHERS, English, Scientific journal
  DOI：https://doi.org/10.1007/s10753-014-9952-8
  DOI ID：10.1007/s10753-014-9952-8, ISSN：0360-3997, eISSN：1573-2576, PubMed ID：25008148, Web of Science ID：WOS:000350553800001
• GADD34 inhibits activation-induced apoptosis of macrophages through enhancement of autophagy　　　　　　　　　　　　　　　
  Sachiko Ito; Yuriko Tanaka; Reina Oshino; Keiko Aiba; Suganya Thanasegaran; Naomi Nishio; Ken-ichi Isobe
  SCIENTIFIC REPORTS, Volume：5, First page：8327, Feb. 2015, [Reviewed]
  Autophagy is a common physiological function in all eukaryotes. The process is induced by depletion of nutrients including amino acids. GADD34 is expressed following DNA damage, ER stresses and amino acid deprivation. Here, we investigated the effects of GADD34 on autophagy and cell activation in macrophages. The deprivation of tyrosine and cysteine markedly induced the expression of GADD34 in macrophages. LPS stimulation combined with tyrosine/cysteine-deprivation initially activated macrophages, but then shifted to cell death in late phase of stimulation. When LPS stimulation was combined with tyrosine/cysteine-deprivation, a deficiency of GADD34 enhanced cell activation signaling such as Src-family, Erk1/2, p38MAPKand Akt. In the late phase of stimulation, a deficiency of GADD34 increased apoptosis more than that in wild-type macrophages. Further we found that mTOR-S6K signaling was highly enhanced in GADD34-deficient macrophages compared with wild-type cells when cells were treated by LPS combined with tyrosine/cysteine-deprivation. LC3-II was increased by LPS stimulation combined with tyrosine/cysteine-deprivation. Defective GADD34 reduced LC3-II and autophagosome formation induced by LPS-stimulation and tyrosine/cysteine-deprivation compared with that seen in wild-type macrophages. These results indicates that GADD34 enhances autophagy and suppresses apoptosis stimulated by LPS combined with amino acid deprivation through regulation of mTOR signaling pathway in macrophages.
  NATURE PUBLISHING GROUP, English, Scientific journal
  DOI：https://doi.org/10.1038/srep08327
  DOI ID：10.1038/srep08327, ISSN：2045-2322, PubMed ID：25659802, Web of Science ID：WOS:000348990600016
• Reprint of "iPSCs, aging and age-related diseases"　　　　　　　　　　　　　　　
  Ken-Ichi Isobe; Zhao Cheng; Naomi Nishio; Thanasegan Suganya; Yuriko Tanaka; Sachiko Ito
  NEW BIOTECHNOLOGY, Volume：32, Number：1, First page：169, Last page：179, Jan. 2015, [Reviewed]
  Human histocompatibility antigens are quite heterogeneous and promote the rejection of transplanted tissue. Recent advances in stem cell research that enable the use of a patient's own stem cells for transplantation are very important because rejection could be avoided. In particular, Yamanaka's group in Japan gave new hope to patients with incurable diseases when they developed induced murine pluripotent stem cells (iPSCs) in 2006 and human iPSCs in 2007. Whereas embryonic stem cells (ESCs) are derived from the inner cell mass and are supported in culture by LIF, iPSCs are derived from fetal or adult somatic cells. Through the application of iPSC technology, adult somatic cells can develop a pluripotent state. One advantage of using iPSCs instead of ESCs in regenerative medicine is that (theoretically) immune rejection could be avoided, although there is some debate about immune rejection of a patient's own iPSCs. Many diseases occur in elderly patients. In order to use regenerative medicine with the elderly, it is important to demonstrate that iPSCs can indeed be generated from older patients. Recent findings have shown that iPSCs can be established from aged mice and aged humans. These iPSCs can differentiate to cells from all three germ layers. However, it is not known whether iPSCs from aged mice or humans show early senescence. Before clinical use of iPSCs, issues related to copy number variation, tumorigenicity and immunogenicity must be resolved. It is particularly important that researchers have succeeded in generating iPSCs that have differentiated to somatic cells related to specific diseases of the elderly, including atherosclerosis, diabetes, Alzheimer's disease and Parkinson's disease. These efforts will facilitate the use of personalized stem cell transplantation therapy for currently incurable diseases.
  ELSEVIER SCIENCE BV, English
  DOI：https://doi.org/10.1016/j.nbt.2014.11.002
  DOI ID：10.1016/j.nbt.2014.11.002, ISSN：1871-6784, eISSN：1876-4347, PubMed ID：25479728, Web of Science ID：WOS:000347507800021
• Enhancement of the Acrolein-Induced Production of Reactive Oxygen Species and Lung Injury by GADD34　　　　　　　　　　　　　　　
  Yang Sun; Sachiko Ito; Naomi Nishio; Yuriko Tanaka; Nana Chen; Lintao Liu; Ken-ichi Isobe
  OXIDATIVE MEDICINE AND CELLULAR LONGEVITY, Volume：2015, First page：170309, 2015, [Reviewed]
  Chronic obstructive pulmonary disease (COPD) is characterized by lung destruction and inflammation. As a major compound of cigarette smoke, acrolein plays a critical role in the induction of respiratory diseases. GADD34 is known as a growth arrest and DNA damage-related gene, which can be overexpressed in adverse environmental conditions. Here we investigated the effects of GADD34 on acrolein-induced lung injury. The intranasal exposure of acrolein induced the expression of GADD34, developing the pulmonary damage with inflammation and increase of reactive oxygen species (ROS). Conversely, the integrality of pulmonary structure was preserved and the generation of ROS was reduced in GADD34-knockout mice. Acrolein-induced phosphorylation of eIF2 alpha in GADD34-knockout epithelial cells by shRNA protected cell death by reducing misfolded protein-caused oxidative stress. These data indicate that GADD34 participates in the development of acrolein-induced lung injury.
  HINDAWI PUBLISHING CORP, English, Scientific journal
  DOI：https://doi.org/10.1155/2015/170309
  DOI ID：10.1155/2015/170309, ISSN：1942-0900, eISSN：1942-0994, PubMed ID：25821552, Web of Science ID：WOS:000351601500001
• iPSCs, aging and age-related diseases　　　　　　　　　　　　　　　
  Ken-ichi Isobe; Zhao Cheng; Naomi Nishio; Thanasegan Suganya; Yuriko Tanaka; Sachiko Ito
  NEW BIOTECHNOLOGY, Volume：31, Number：5, First page：411, Last page：421, Sep. 2014, [Reviewed]
  Human histocompatibility antigens are quite heterogeneous and promote the rejection of transplanted tissue. Recent advances in stem cell research that enable the use of a patient's own stem cells for transplantation are very important because rejection could be avoided. In particular, Yamanaka's group in Japan gave new hope to patients with incurable diseases when they developed induced murine pluripotent stem cells (iPSCs) in 2006 and human iPSCs in 2007. Whereas embryonic stem cells (ESCs) are derived from the inner cell mass and are supported in culture by LIF, iPSCs are derived from fetal or adult somatic cells. Through the application of iPSC technology, adult somatic cells can develop a pluripotent state. One advantage of using iPSCs instead of ESCs in regenerative medicine is that (theoretically) immune rejection could be avoided, although there is some debate about immune rejection of a patient's own iPSCs. Many diseases occur in elderly patients. In order to use regenerative medicine with the elderly, it is important to demonstrate that iPSCs can indeed be generated from older patients. Recent findings have shown that iPSCs can be established from aged mice and aged humans. These iPSCs can differentiate to cells from all three germ layers. However, it is not known whether iPSCs from aged mice or humans show early senescence. Before clinical use of iPSCs, issues related to copy number variation, tumorigenicity and immunogenicity must be resolved. It is particularly important that researchers have succeeded in generating iPSCs that have differentiated to somatic cells related to specific diseases of the elderly, including atherosclerosis, diabetes, Alzheimer's disease and Parkinson's disease. These efforts will facilitate the use of personalized stem cell transplantation therapy for currently incurable diseases.
  ELSEVIER SCIENCE BV, English, Scientific journal
  DOI：https://doi.org/10.1016/j.nbt.2014.04.004
  DOI ID：10.1016/j.nbt.2014.04.004, ISSN：1871-6784, eISSN：1876-4347, PubMed ID：24784583, Web of Science ID：WOS:000341314700003
• Acrolein induced both pulmonary inflammation and the death of lung epithelial cells　　　　　　　　　　　　　　　
  Yang Sun; Sachiko Ito; Naomi Nishio; Yuriko Tanaka; Nana Chen; Ken-ichi Isobe
  TOXICOLOGY LETTERS, Volume：229, Number：2, First page：384, Last page：392, Sep. 2014, [Reviewed]
  Acrolein, a compound found in cigarette smoke, is a major risk factor for respiratory diseases. Previous research determined that both acrolein and cigarette smoke produced reactive oxygen species (ROS). As many types of pulmonary injuries are associated with inflammation, this study sought to ascertain the extent to which exposure to acrolein advanced inflammatory state in the lungs. Our results showed that intranasal exposure of mice to acrolein increased CD11c(+)F4/80(high) macrophages in the lungs and increased ROS formation via induction of NF-kappa B signaling. Treatment with acrolein activated macrophages and led to their increased production of ROS and expression of several key pro-inflammatory cytokines. In in vitro studies, acrolein treatment of bone marrow-derived GM-CSF-dependent immature macrophages (GM-IMs), activated the cells and led to their increased production of ROS and expression of several key pro-inflammatory cytokines. Acrolein treatment of macrophages induced apoptosis of lung epithelial cells. Inclusion of an inhibitor of ROS formation markedly decreased acrolein-mediated macrophage activation and reduced the extent of epithelial cell death. These results indicate that acrolein can cause lung damage, in great part by mediating the increased release of proinflammatory cytokines/factors by macrophages. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
  ELSEVIER IRELAND LTD, English, Scientific journal
  DOI：https://doi.org/10.1016/j.toxlet.2014.06.021
  DOI ID：10.1016/j.toxlet.2014.06.021, ISSN：0378-4274, eISSN：1879-3169, PubMed ID：24999835, Web of Science ID：WOS:000340045700008
• Negative Regulation of GADD34 on Myofibroblasts during Cutaneous Wound Healing　　　　　　　　　　　　　　　
  Lintao Liu; Naomi Nishio; Sachiko Ito; Yuriko Tanaka; Ken-ichi Isobe
  BIOMED RESEARCH INTERNATIONAL, Volume：2014, First page：137049, 2014, [Reviewed]
  The growth arrest and DNA damage-inducible protein, GADD34, has been proved to be involved in TGF-beta signaling pathway and correlates with cell death, which are two important mechanisms in regulating myofibroblast differentiation and apoptosis during tissue repair. But roles of GADD34 in myofibroblasts differentiation and apoptosis remain unknown. To investigate the function of GADD34 in these processes, we subjected WT and GADD34(-/)-mice to dermal wound healing. Here we show that GADD34(-/)-mice exhibited accelerated wound closure compared withWT mice. In addition, GADD34(-/)-mice showed increased number of myofibroblasts, elevated collagen production, and decreased cell apoptosis during wound healing. Moreover, we found that ADD34(-/)-mice showed increased phosphorylation of Smad3 and lower level of cleaved caspase-3. Thus these results indicate that GADD34 appears to suppress myofibroblast differentiation through inhibiting Smad3-dependent TGF-beta signal pathway and promote its apoptosis by activating caspase-3 pathway.
  HINDAWI PUBLISHING CORPORATION, English, Scientific journal
  DOI：https://doi.org/10.1155/2014/137049
  DOI ID：10.1155/2014/137049, ISSN：2314-6133, eISSN：2314-6141, PubMed ID：25210702, Web of Science ID：WOS:000347824500001
• [Accuracy estimation of non-invasive X-ray output analyzer].　　　　　　　　　　　　　　　
  Ioka Y; Ariga E; Nishio N; Ooshima T
  Nihon Hoshasen Gijutsu Gakkai zasshi, Volume：69, Number：10, First page：1153, Last page：1160, Oct. 2013, [Reviewed]
  This study estimated the accuracy of an X-ray analyzer by comparing it with an ionization chamber and a tube voltage current meter, and investigated whether it was usable as a substitute for a reference meter for output measurements for quality control purposes. The X-ray output analyzer used was a Piranha (RTI Electronics), a non-invasive instrument. The two subjects of measurements were as follows: the tube voltage, exposure, and half-value layer used in ordinary X-ray radiographic system equipment, and the exposure and half-value layer in X-ray equipment for mammographic systems. The results for a conventional radiographic system showed the error rates for tube voltage, exposure, and half value layer to be within ±1.0%, ±1.8%, and ±4.3%, respectively. The Piranha is not influenced by the dependence of the beam quality in a range of the tube voltage in clinical use. In X-ray equipment for mammographic system results, error rates for exposure and half value layer were ±2.2% and within ±4.0%, respectively. We conclude that it is possible to use the Piranha as an alternative reference meter for quality control of X-ray equipment for typical radiographic and mammographic systems.
  Japanese Society of Radiological Technology, Japanese
  DOI：https://doi.org/10.6009/jjrt.2013_JSRT_69.10.1153
  DOI ID：10.6009/jjrt.2013_JSRT_69.10.1153, ISSN：0369-4305, CiNii Articles ID：10031196868, CiNii Books ID：AN00197784, PubMed ID：24140904
• Characteristics of cardiac aging in C57BL/6 mice　　　　　　　　　　　　　　　
  Zhao Cheng; Sachiko Ito; Naomi Nishio; Suganya Thanasegaran; He Fang; Ken-ichi Isobe
  EXPERIMENTAL GERONTOLOGY, Volume：48, Number：3, First page：341, Last page：348, Mar. 2013, [Reviewed]
  The specific processes that cause aging of the cardiac tissue remain elusive. C57BL/6 (B6) mice are commonly used for investigating age-related diseases in mammals. We thus sought to evaluate the cardiac aging process in B6 mice. Cardiac tissues from the newborn (B6 NB), 2 month-old (B6 2M) and 21-27 month-old B6 mice (B6 aged) were used for the investigation. Several age-related cellular processes were evaluated, including telomere shortening, changes in p53 and p16 expression, changes in mitochondria DNA expression and DNA deletion, and alteration of mitochondria. We found that the aging of the B6 mice cardiac tissue is associated with the maintenance of telomere length, increased expression of p53 and p16, mild changes in mitochondrial DNA expression but widespread DNA deletion, and significant alterations of the mitochondrial ultrastructure within the cardiac tissue. The results of our studies suggest that mitochondrial DNA deletions, which affect the mitochondrial ultrastructure, cytochrome C oxidase activity, and p53 expression, are significantly associated with cardiac aging and may be a source of age-related heart failure. (C) 2013 Elsevier Inc. All rights reserved.
  PERGAMON-ELSEVIER SCIENCE LTD, English, Scientific journal
  DOI：https://doi.org/10.1016/j.exger.2013.01.005
  DOI ID：10.1016/j.exger.2013.01.005, ISSN：0531-5565, PubMed ID：23337778, Web of Science ID：WOS:000315683800006
• iPS cell sheets created by a novel magnetite tissue engineering method for reparative angiogenesis　　　　　　　　　　　　　　　
  Tetsutaro Kito; Rei Shibata; Masakazu Ishii; Hirohiko Suzuki; Tatsuhito Himeno; Yoshiyuki Kataoka; Yumiko Yamamura; Takashi Yamamoto; Naomi Nishio; Sachiko Ito; Yasushi Numaguchi; Tohru Tanigawa; Jun K. Yamashita; Noriyuki Ouchi; Hiroyuki Honda; Kenichi Isobe; Toyoaki Murohara
  SCIENTIFIC REPORTS, Volume：3, First page：1418, Mar. 2013, [Reviewed]
  Angiogenic cell therapy represents a novel strategy for ischemic diseases, but some patients show poor responses. We investigated the therapeutic potential of an induced pluripotent stem (iPS) cell sheet created by a novel magnetite tissue engineering technology (Mag-TE) for reparative angiogenesis. Mouse iPS cell-derived Flk-1(+) cells were incubated with magnetic nanoparticle-containing liposomes (MCLs). MCL-labeled Flk-1(+) cells were mixed with diluted extracellular matrix (ECM) precursor and a magnet was placed on the reverse side. Magnetized Flk-1(+) cells formed multi-layered cell sheets according to magnetic force. Implantation of the Flk-1(+) cell sheet accelerated revascularization of ischemic hindlimbs relative to the contralateral limbs in nude mice as measured by laser Doppler blood flow and capillary density analyses. The Flk-1(+) cell sheet also increased the expressions of VEGF and bFGF in ischemic tissue. iPS cell-derived Flk-1(+) cell sheets created by this novel Mag-TE method represent a promising new modality for therapeutic angiogenesis.
  NATURE PUBLISHING GROUP, English, Scientific journal
  DOI：https://doi.org/10.1038/srep01418
  DOI ID：10.1038/srep01418, ISSN：2045-2322, PubMed ID：23475393, Web of Science ID：WOS:000315852100004
• No immunogenicity of IPS cells in syngeneic host studied by in vivo injection and 3D scaffold experiments.　　　　　　　　　　　　　　　
  Suganya Thanasegaran; Zhao Cheng; Sachiko Ito; Naomi Nishio; Ken-ichi Isobe
  BioMed research international, Volume：2013, First page：378207, Last page：378207, 2013, [Reviewed], ［International magazine］
  Induced Pluripotent Stem Cells (IPSCs) open the great possibility to employ patient's own tissue to the previously incurable diseases. However these cells can be used in cell therapy only if they are not rejected when transplanted back into the syngeneic host. We found that the injection of iPSCs derived from different ages of mice into syngeneic C57BL/6 mice produced teratoma and was not rejected. Then we cultured iPSCs and myeloid differentiated iPSCs in three-dimensional porous scaffold and transplanted to C57BL/6 mice and BALB/C mice. After transplantation, we could observe the cell density inside the scaffold increased rapidly in syngeneic mice compared to the allogeneic mice indicating the favorable conditions supporting the growth of iPSCs in vivo. Unlike the allogeneic counterpart, we could not observe few infiltrating T cells inside the scaffold of syngeneic mice. These results contribute to the optimistic view of iPSCs for regenerative medicine in near future.
  English, Scientific journal
  DOI：https://doi.org/10.1155/2013/378207
  DOI ID：10.1155/2013/378207, ISSN：2314-6133, PubMed ID：23691499, PubMed Central ID：PMC3652117
• Establishment of self-renewable GM-CSF-dependent immature macrophages in vitro from murine bone marrow.　　　　　　　　　　　　　　　
  Sachiko Ito; Yuriko Tanaka; Naomi Nishio; Suganya Thanasegaran; Ken-Ichi Isobe
  PloS one, Volume：8, Number：10, First page：e76943, 2013, [Reviewed], ［International magazine］
  Macrophages play a key role in the innate immune system. Macrophages are thought to originate from hematopoietic precursors or the yolk sac. Here, we describe the in vitro establishment of self-renewable GM-CSF-dependent immature macrophages (GM-IMs) from murine bone marrow (BM). GM-IMs grow continuously in vitro in conditioned medium containing GM-CSF. The immunophenotype of GM-IMs is F4/80(high) CD11b(high) CD11c(low) Ly6C(low). By comparing gene expression in GM-IMs and BM dendritic cells, we found that GM-IMs expressed lower levels of chemokines, cytokines and their receptors. GM-IMs are round in shape, attach loosely to non-coated culture dishes and have a marked phagocytic capacity. These results indicate that GM-IMs are macrophage precursor cells. Following stimulation with LPS, monocyte-like GM-IMs converted to flat macrophage-like cells that tightly adhered to non-coated culture dishes and produced pro-inflammatory cytokines TNFα, IL-6 and IL-1β. These results indicated that GM-IMs differentiated to M1 pro-inflammatory macrophages. This was confirmed by stimulation of GM-IMs with IFNγ, an inducer of M1 markers. GM-IMs showed enhanced expression of M2 macrophage markers such as Arg1 and Retnla following stimulation by Th2 cytokines IL-4 and IL-13. When GM-IMs were injected into mice at sites of wounding, wound repair was enhanced. These results indicate that GM-IMs can differentiate to M2 macrophages. When GM-IMs were injected into clodronate-treated mice, they induced resident macrophage proliferation by producing M-CSF. In conclusion we have established self-renewable GM-CSF-dependent immature macrophages in vitro from murine BM, which differentiate to M1 or M2 macrophages.
  English, Scientific journal
  DOI：https://doi.org/10.1371/journal.pone.0076943
  DOI ID：10.1371/journal.pone.0076943, PubMed ID：24124601, PubMed Central ID：PMC3790761
• Comparative Angiogenic Activities of Induced Pluripotent Stem Cells Derived from Young and Old Mice　　　　　　　　　　　　　　　
  Hirohiko Suzuki; Rei Shibata; Tetsutaro Kito; Takashi Yamamoto; Masakazu Ishii; Naomi Nishio; Sachiko Ito; Ken-ichi Isobe; Toyoaki Murohara
  PLOS ONE, Volume：7, Number：6, First page：e39562, Jun. 2012, [Reviewed]
  Advanced age is associated with decreased stem cell activity. However, the effect of aging on the differentiation capacity of induced pluripotent stem (iPS) cells into cardiovascular cells has not been fully clarified. We investigated whether iPS cells derived from young and old mice are equally capable of differentiating into vascular progenitor cells, and whether these cells regulate vascular responses in vivo. iPS cells from mouse embryonic fibroblasts (young) or 21 month-old mouse bone marrow (old) were used. Fetal liver kinase-1 positive (Flk-1(+)) cells, as a vascular progenitor marker, were induced after 3 to 4 days of culture from iPS cells derived from young and old mice. These Flk-1(+) cells were sorted and shown to differentiate into VE-cadherin(+) endothelial cells and alpha-SMA(+) smooth muscle cells. Tube-like formation was also successfully induced in both young and old murine Flk-1(+) cells. Next, hindlimb ischemia was surgically induced, and purified Flk-1(+) cells were directly injected into ischemic hindlimbs of nude mice. Revascularization of the ischemic hindlimb was significantly accelerated in mice transplanted with Flk-1(+) cells derived from iPS cells from either young or old mice, as compared to control mice as evaluated by laser Doppler blood flowmetry. The degree of revascularization was similar in the two groups of ischemic mice injected with iPS cell-derived Flk-1(+) cells from young or old mice. Transplantation of Flk-1(+) cells from both young and old murine iPS cells also increased the expression of VEGF, HGF and IGF mRNA in ischemic tissue as compared to controls. iPS cell-derived Flk-1(+) cells differentiated into vascular progenitor cells, and regulated angiogenic vascular responses both in vitro and in vivo. These properties of iPS cells derived from old mice are essentially the same as those of iPS cells from young mice, suggesting the functionality of generated iPS cells themselves to be unaffected by aging.
  PUBLIC LIBRARY SCIENCE, English, Scientific journal
  DOI：https://doi.org/10.1371/journal.pone.0039562
  DOI ID：10.1371/journal.pone.0039562, ISSN：1932-6203, PubMed ID：22761825, Web of Science ID：WOS:000305825800043
• Autophagic activity in thymus and liver during aging　　　　　　　　　　　　　　　
  Mohammad Nizam Uddin; Naomi Nishio; Sachiko Ito; Haruhiko Suzuki; Ken-ichi Isobe
  AGE, Volume：34, Number：1, First page：75, Last page：85, Feb. 2012, [Reviewed]
  Impaired or deficient autophagy is believed to cause or contribute to aging, as well as several age-related pathologies. Thymic epithelial cells had a high constitutive level of autophagy. The autophagic process may play a supporting role or even a crucial role in the presentation of self-Ags in the thymus to shape the T-cell repertoires. Autophagic activity in the liver is important for the balance of energy and nutrients for basic cell functions. The abundance of autophagic structure in both cortical and medullary thymic epithelial cells and liver with mouse age has not been examined in detail. Here, we demonstrated that the architecture of mouse thymus and liver markedly changed with age. We found that the expression of LC3 detected by immunofluorescence and Western blot analysis was greatly decreased in thymus and liver of 12-month-old mice. The same level of reduction was observed in thymus and liver of 24-month-old mice. Ultrastructure analysis by an electron microscope revealed that the number of autophagic structure/vacuole in total thymic epithelial cells and hepatocytes decrease with age. The age-related decrease of autophagic structure in thymic epithelial cells may cause the reduction of immunocompetent T-cell pool in aged mice. The age-related decrease of autophagy in liver may induce accumulation of cellular materials in liver of aged mice.
  SPRINGER, English, Scientific journal
  DOI：https://doi.org/10.1007/s11357-011-9221-9
  DOI ID：10.1007/s11357-011-9221-9, ISSN：0161-9152, PubMed ID：21387084, Web of Science ID：WOS:000299294100007
• 3D-aggregated dermal stem cells with partial-pluripotency　　　　　　　　　　　　　　　
  Masaki Kondo; Hideki Kamiya; Tetsuji Okawa; Sachiko Ito; Naomi Nishio; Tatsuhito Himeno; Yutaka Oiso; Jiro Nakamura; Ken-Ichi Isobe
  2012 International Symposium on Micro-NanoMechatronics and Human Science, MHS 2012, First page：156, Last page：159, 2012, [Reviewed]
  Current stem cell therapy is expected to be an efficient method for replacement of lost cells in many intractable diseases, and several types of cells are considered candidate sources including iPSCs and ESCs. However, these cells still need to elucidate several serious problems. We are trying to establish safe pluripotent stem cells for early use to clinic. Here we have identified and established 3D spheroidal dermal stem cells, which possess partial pluripotency. We cultured the dermal cells from GFP carrying C57BL/6 male mice. After their second passage, we dissociate and quickly aggregate these cells and cultured on U-bottom 96-well low-cell-adhesion plates. The cells self aggregated in 3D structure. We examined pluripotent markers. We found that 3D-aggregated sphere expressed Nanog, Oct4, Sox2 and SSEA-1 protein. We named these 3D sphere as spheroidal dermal stem cells (sDSCs). Then we tried to differentiate sDSCs to ectodermal-lineage cells on PA6 cells. We found the appearance of neuron-like cells, having the neuronal markers such as Neurofilament and S-100b positive. For endodermal-lineage cell induction, sDSCs were cultured on collagen-coated dishes with FBS and Activin A. The differentiated cells express α-fetoprotein and Pdx-1. This study shows that sDSCs would be the low risk and potential applications of cell transplantation therapy and be able to breakthrough in stem cell treatment. © 2012 IEEE.
  English, International conference proceedings
  DOI：https://doi.org/10.1109/MHS.2012.6492401
  DOI ID：10.1109/MHS.2012.6492401, SCOPUS ID：84876543437
• Establishment of neutrophil-lineage stem cells from C57BL/6 mice.　　　　　　　　　　　　　　　
  Naomi Nishio; Sachiko Ito; Yuriko Tanaka; Ken-ichi Isobe
  2012 INTERNATIONAL SYMPOSIUM ON MICRO-NANOMECHATRONICS AND HUMAN SCIENCE (MHS), First page：160, Last page：163, 2012, [Reviewed]
  Neutrophils are key effector player of the innate immune system that are rapidly recruited to infected tissues to clear pathogens. Neutrophils also have the capacity to engulf damaged tissue cells to clear, although at the same time neutrophil granules including myeloperioxidase, elastase and ROS may damage tissues. We have shown previously that neutrophils help to repair wounded tissue. However, neutrophils are already differentiated cells, which cannot divide. For future personalized stem cells therapy, it is important to establish neutrophil-precursor cells, which will differentiate to mature neutrophils to engulf pathogens or damaged cells by the transplantation in vivo. First we characterized 32Dcl3 cell line, which has been established previously by WEHI-3 conditional medium. These cell lines differentiated to neutrophils by the stimulation of G-CSF. Then we established similar clone from C57BL/6 mice by culturing bone marrow cells in WEHI-3 conditional medium. We could obtain a neutrophil-precursor cell line (B6NPC), which grows more than six months in WEHI-3 conditional medium. By FACS analysis, we found that both 32Dcl3 and B6NPC had GR-1, CD11b and F4/80 cell surface markers, which are myeloid-lineage markers. However, these cells also have early T cell markers, which fit the hypothesis of myeloid-based model of hematopoietic cell differentiation. By the transplantation of B6NPC to syngeneic mice at the same time of wounding, we found early recovery from wound healing.
  IEEE, English, International conference proceedings
  ISSN：2474-378X, eISSN：2474-3798, Web of Science ID：WOS:000318860500034
• Possibility to use iPS-technology in age-related diseases　　　　　　　　　　　　　　　
  Zhao Cheng; Sachiko Ito; Naomi Nishio; Thanasegaran Suganya; Ken-ichi Isobe
  2012 INTERNATIONAL SYMPOSIUM ON MICRO-NANOMECHATRONICS AND HUMAN SCIENCE (MHS), First page：116, Last page：120, 2012, [Reviewed]
  When applying the iPSCs for regenerative therapy in elderly patients, it is necessary to establish the iPSCs from elderly patients themselves then differentiate the iPSCs to specific cell types for transplantation treatments. The mouse is a perfect model to study aging in mammals. Bone marrow (BM) cells from aged C57BL/6 mice (15 months to 21 months) were cultured with granulocyte macrophage-colony stimulating factor (GM-CSF). The efficiency to produce iPSCs from aged mice BM cells was lower than that of young mice. We established several clones of iPSCs from aged mice. All the established clones have pluripotent markers. Aged-iPSCs could differentiate to three germ layers in vitro and made teratoma in vivo. We are currently making chimeric mice between Aged-iPSC-1 and ICR mice.
  IEEE, English, International conference proceedings
  ISSN：2474-378X, eISSN：2474-3798, Web of Science ID：WOS:000318860500026
• Aging in the mouse and perspectives of rejuvenation through induced pluripotent stem cells (iPSCs).　　　　　　　　　　　　　　　
  Ken-ichi Isobe; Zhao Cheng; Sachiko Ito; Naomi Nishio
  Results and problems in cell differentiation, Volume：55, First page：413, Last page：27, 2012, [Reviewed], ［International magazine］
  The mouse is a perfect model to study aging in mammals. It has a relatively short life span and genetic manipulations in this species are well established. Most interestingly, the mouse is a fantastic tool to produce stem cells. Forced expression of only four transcription factors (Oct3/4, Sox2, Klf4, and c-Myc) in murine and human somatic cells resets the expression of genes that are characteristic of differentiated cells and consequently induces the formation of pluripotent stem cells (iPSCs). This technology opens new and exciting possibilities in medical research, especially personalized cell therapies for treating human disease. To treat damaged tissues or repair organs in elderly patients, it will be necessary to establish iPSCs from their tissues. To determine the feasibility of using this technology with elderly patients, we asked whether it was indeed possible to establish iPSCs from the tissues of aged mice and to differentiate them to tissue cells. We succeeded in establishing iPSC clones using bone marrow (BM) from 21-month-old EGFP-C57BL/6 mice, which had been cultured for 4 days in the presence of granulocyte macrophage-colony stimulating factor (GM-CSF). Our iPSCs from aged mice (aged iPSCs) and those from mouse embryonic fibroblasts (MEFs) strongly expressed SSEA-1 and Pou5f1, and showed strong alkaline phosphatase (AP) activity. Our aged iPSCs made teratomas when injected into the back skin of syngeneic mice, and differentiated to tissue cells of three germ lines in vitro. Further experiments to make chimeric mice and germ line cells will determine whether the aged iPSCs possess the properties of much younger cells and are capable of regenerating aged mice.
  English, Scientific journal
  DOI：https://doi.org/10.1007/978-3-642-30406-4_21
  DOI ID：10.1007/978-3-642-30406-4_21, ISSN：0080-1844, PubMed ID：22918818
• Dextran sulphate sodium increases splenic Gr1+CD11b+ cells which accelerate recovery from colitis following intravenous transplantation　　　　　　　　　　　　　　　
  R. Zhang; S. Ito; N. Nishio; Z. Cheng; H. Suzuki; K. I. Isobe
  Clinical and Experimental Immunology, Volume：164, Number：3, First page：417, Last page：427, Jun. 2011, [Reviewed]
  Summary: While Gr1+CD11b+ cells are known to regulate immune responses and accumulate in most cancer tissues, the function of Gr1+CD11b+ cells in inflammation is poorly understood. We investigated the role of Gr1+CD11b+ cells in a dextran sulphate sodium (DSS)-treated mouse model of ulcerative colitis (UC). C57BL/6 mice were treated with 2% DSS in drinking water for 5 days. Disease progression and recovery were assessed by body weight, disease activity index score (DAI) score and colon length. Splenic Gr1+CD11b+ cell number was greatly increased during the recovery phase of DSS-induced colitis. DSS-derived splenic Gr1+CD11b+ cells were administered intravenously to recipient (C57BL/6) mice during the early phase of DSS treatment. The transplanted splenic DSS-induced Gr1+CD11b+ cells improved DSS-induced colitis and promoted efficient colonic mucosal healing. We found that the CD11b+ single positive cells increased in the course of DSS-induced colitis in lamina propria. The transplantation of splenic Gr1+CD11b+ cells induced feedback suppression of myeloid-lineage cell development. Namely, the transplantation of splenic Gr1+CD11b+ cells greatly suppressed the migration of CD11b+ single positive cells to the lamina propria. Further, transplantation of Gr-1+CD11b+ cells greatly suppressed the increase of the same population, especially during the late phase of DSS colitis both in spleen and bone marrow. © 2011 The Authors. Clinical and Experimental Immunology © 2011 British Society for Immunology.
  English, Scientific journal
  DOI：https://doi.org/10.1111/j.1365-2249.2011.04374.x
  DOI ID：10.1111/j.1365-2249.2011.04374.x, ISSN：0009-9104, PubMed ID：21413942, SCOPUS ID：79955417245
• Establishment of induced pluripotent stem cells from aged mice using bone marrow-derived myeloid cells　　　　　　　　　　　　　　　
  Zhao Cheng; Sachiko Ito; Naomi Nishio; Hengyi Xiao; Rong Zhang; Haruhiko Suzuki; Yayoi Okawa; Toyoaki Murohara; Ken-ichi Isobe
  JOURNAL OF MOLECULAR CELL BIOLOGY, Volume：3, Number：2, First page：91, Last page：98, Apr. 2011, [Reviewed]
  If induced pluripotent stem (iPS) cells are to be used to treat damaged tissues or repair organs in elderly patients, it will be necessary to establish iPS cells from their tissues. To determine the feasibility of using this technology with elderly patients, we asked if it was indeed possible to establish iPS cells from the bone marrow (BM) of aged mice. BM cells from aged C57BL/6 mice carrying the green fluorescence protein (GFP) gene were cultured with granulocyte macrophage-colony stimulating factor (GM-CSF) for 4 days. Four factors (Oct3/4, Sox2, Klf4 and c-Myc) were introduced into the BM-derived myeloid (BM-M) cells. The efficiency of generating iPS cells from aged BM cultured in GM-CSF was low. However, we succeeded in obtaining BM-M-iPS cells from aged C57BL/6 mice, which carried GFP. Our BM-M-iPS cells expressed SSEA-1 and Pou5f1 and were positive for alkaline phosphatase staining. The iPS cells did make teratoma with three germ layers following injection into syngeneic C57BL/6 mice, and can be differentiated to three germ layers in vitro. By co-culturing with OP9, the BM-M-iPS cells can be differentiated to the myeloid lineage. The differentiated BM-M-iPS cells proliferated well in the presence of GM-CSF, and lost expression of Nanog and Pou5f1, at least in part, due to methylation of their promoters. On the contrary, Tnf and Il1b gene expression was upregulated and their promoters were hypomethylated.
  OXFORD UNIV PRESS, English, Scientific journal
  DOI：https://doi.org/10.1093/jmcb/mjq044
  DOI ID：10.1093/jmcb/mjq044, ISSN：1674-2788, eISSN：1759-4685, PubMed ID：21228011, Web of Science ID：WOS:000289306800004
• Gadd34 induces autophagy through the suppression of the mTOR pathway during starvation　　　　　　　　　　　　　　　
  Mohammad Nizam Uddin; Sachiko Ito; Naomi Nishio; Thanasegaran Suganya; Ken-ichi Isobe
  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, Volume：407, Number：4, First page：692, Last page：698, Apr. 2011, [Reviewed]
  Several types of cellular stress induce expression of growth arrest and DNA damage protein 34 (Gadd34). Autophagy occurs under both basal conditions and conditions of stress, such as starvation. Gadd34 and autophagy are both induced under starvation conditions. In this study we found that starvation induced the expression of Gadd34, reduced mTOR activity, and induced autophagy in wild type mice, but not Gadd34 KO mice. Gadd34 bound to and dephosphorylated pTSC2 at Thr1462. Dephosphorylation of TSC2 during the starvation time period leads to the suppression of mTOR, which is a potent inhibitor of autophagy. We concluded that starvation-induced Gadd34 suppresses mTOR and, thereby, induces autophagy. (C) 2011 Elsevier Inc. All rights reserved.
  ACADEMIC PRESS INC ELSEVIER SCIENCE, English, Scientific journal
  DOI：https://doi.org/10.1016/j.bbrc.2011.03.077
  DOI ID：10.1016/j.bbrc.2011.03.077, ISSN：0006-291X, PubMed ID：21439266, Web of Science ID：WOS:000290240500012
• Differentiation of induced pluripotent stem cells to thymic epithelial cells by phenotype　　　　　　　　　　　　　　　
  Yuta Inami; Tohru Yoshikai; Sachiko Ito; Naomi Nishio; Haruhiko Suzuki; Hidetoshi Sakurai; Ken-Ichi Isobe
  IMMUNOLOGY AND CELL BIOLOGY, Volume：89, Number：2, First page：314, Last page：321, Feb. 2011, [Reviewed]
  Thymic epithelial cells (TECs) are present in both cortical and medullary thymic areas, and have crucial roles in functional T-cell development. In this study, we studied the differentiation of induced pluripotent stem cells (iPSCs) to TEC. When iPSC were cultured for 4 days in collagen IV-coated dishes in the presence of both activin A and lithium chloride (LiCl), the cells differentiated to definitive endoderm through mesendoderm. Further treatment with Fgf8 followed by Fgf7, Fgf10 and BMP4 differentiated iPSC to thymic epithelial progenitor cells (TEPCs) by phenotype. Gene expression of Hoxa3, Pax1 and Pax9 was observed and cell surface proteins EpCAM1 and MTS24 were detected at day 14 of iPSC differentiation. TEPCs differentiated to medullary TECs (mTECs) by phenotype following the addition of receptor activator nuclear factor B ligand with LiCl. Thus, we successfully induced efficient differentiation from mouse iPSC to TEPCs and mTEC by phenotype using chemically defined conditions. Immunology and Cell Biology (2011) 89, 314-321; doi:10.1038/icb.2010.96; published online 3 August 2010
  NATURE PUBLISHING GROUP, English, Scientific journal
  DOI：https://doi.org/10.1038/icb.2010.96
  DOI ID：10.1038/icb.2010.96, ISSN：0818-9641, PubMed ID：20680027, Web of Science ID：WOS:000287445400021
• Up-regulation of Gr1+CD11b+ population in spleen of dextran sulfate sodium administered mice works to repair colitis.　　　　　　　　　　　　　　　
  Rong Zhang; Sachiko Ito; Naomi Nishio; Zhao Cheng; Haruhiko Suzuki; Ken-ichi Isobe
  Inflammation & allergy drug targets, Volume：10, Number：1, First page：39, Last page：46, Feb. 2011, [Reviewed], ［International magazine］
  Dextran sulfate sodium (DSS) is commonly used in rodent IBD models to chemically induce acute intestinal inflammation. The acute course of colitis includes colon tissue damages and recovery from wounded tissues. As skin wound repair was delayed by splenectomy, we asked whether splenectomy would induce the delay of colonic wound healing. In splenectomized mice, body weight recovery, disease score and colon length were delayed. Surprisingly we found a great increase of Gr1+CD11b+ cells in spleen and bone marrow of DSS-administered mice. Anti-Gr-1 antibody treatment worsened the DSS- administered colitis. These results indicate that Gr1+CD11b+ cells induced by DSS worked to repair colon wound healing and repair colitis.
  English, Scientific journal
  ISSN：1871-5281, eISSN：2212-4055, PubMed ID：21184649
• Toxic effects of D-galactose on thymus and spleen that resemble aging　　　　　　　　　　　　　　　
  Mohammad Nizam Uddin; Naomi Nishio; Sachiko Ito; Haruhiko Suzuki; Ken-ichi Isobe
  JOURNAL OF IMMUNOTOXICOLOGY, Volume：7, Number：3, First page：165, Last page：173, Sep. 2010, [Reviewed]
  Continuous low-dose injection of D-galactose induces changes in mice that resemble accelerated aging. As such, these mice have been used as models to study mechanisms of aging. Here, we examined whether repeated (daily, for 60 days) subcutaneous injections (at 50 mg D-galactose/kg) into young adult (i.e., 2-month-old) mice induced changes in key immune system organs that were on par with those associated with aging. The results showed that galactose-treated mice develop histologic changes in their thymic cortical and medullary regions; immunohistochemical analysis revealed unorganized distributions of keratin-5 and keratin-8 proteins in the thymus of these hosts. These histological changes in the thymus of D-galactose-treated mice were also observed in the organs of aged (i.e., 24-month-old control mice); however, in this latter group, these changes were accompanied by a strong infiltration of adipose cells. Galactose-treated mice also evinced alterations within their splenic white and red pulp. Further, ultrastructural analyses of the thymus and spleen of the treated mice revealed increases in irregularly shaped lymphocytes bearing visible pyknosis. It was also seen that levels of autophagy within thymic epithelial cells were greatly decreased in the tissues of the galactose-treated mice, an outcome also seen in aged mice. Lastly, the level of memory T-lymphocytes and percentage of IgM-B220-B-lymphocytes in spleens of the galactose-treated mice were both increased (albeit insignificantly so) relative to values among splenocytes of age-matched control; however, these levels were not clealy as elevated as would be expected in "elderly" mice. Taken together, our results strongly suggest that D-galactose treatment can induce structural changes in the thymus and spleen, and some changes in organ-associated cell phenotypes, that are similar to several effects seen with aging. However, the fact that many endpoints do not appear to be truly reflective of what should be seen in immune system organs/cells of "elderly" mice now calls into question the appropriateness of the use of D-galactose (i.e., is it histologically/immunotoxicologically-proper?) to create age-mimicry in mice.
  INFORMA HEALTHCARE, English, Scientific journal
  DOI：https://doi.org/10.3109/15476910903510806
  DOI ID：10.3109/15476910903510806, ISSN：1547-691X, PubMed ID：20050818, Web of Science ID：WOS:000281919600003
• Therapeutic angiogenesis by transplantation of induced pluripotent stem cell-derived Flk-1 positive cells　　　　　　　　　　　　　　　
  Hirohiko Suzuki; Rei Shibata; Tetsutaro Kito; Masakazu Ishii; Ping Li; Toru Yoshikai; Naomi Nishio; Sachiko Ito; Yasushi Numaguchi; Jun K. Yamashita; Toyoaki Murohara; Kenichi Isobe
  BMC CELL BIOLOGY, Volume：11, First page：72, Sep. 2010, [Reviewed]
  Background: Induced pluripotent stem (iPS) cells are the novel stem cell population induced from somatic cells. It is anticipated that iPS will be used in the expanding field of regenerative medicine. Here, we investigated whether implantation of fetal liver kinase-1 positive (Flk-1(+)) cells derived from iPS cells could improve angiogenesis in a mouse hind limb model of ischemia.
  Results: Flk-1(+) cells were induced from iPS cells after four to five days of culture. Hind limb ischemia was surgically induced and sorted Flk-1(+) cells were directly injected into ischemic hind limbs of athymic nude mice. Revascularization of the ischemic hind limb was accelerated in mice that were transplanted with Flk-1(+) cells compared with control mice, which were transplanted with vehicle, as evaluated by laser Doppler blood flowmetry. Transplantation of Flk-1(+) cells also increased expression of VEGF mRNA in ischemic tissue compared to controls.
  Conclusions: Direct local implantation of iPS cell-derived Flk-1(+) cells would salvage tissues from ischemia. These data indicate that iPS cells could be valuable in the therapeutic induction of angiogenesis.
  BIOMED CENTRAL LTD, English, Scientific journal
  DOI：https://doi.org/10.1186/1471-2121-11-72
  DOI ID：10.1186/1471-2121-11-72, ISSN：1471-2121, PubMed ID：20860813, Web of Science ID：WOS:000283075600001
• GADD34 suppresses wound healing by upregulating expression of myosin IIA　　　　　　　　　　　　　　　
  Chie Tanaka; Sachiko Ito; Naomi Nishio; Yasuhiro Kodera; Hidetoshi Sakurai; Haruhiko Suzuki; Akimasa Nakao; Ken-Ichi Isobe
  TRANSGENIC RESEARCH, Volume：19, Number：4, First page：637, Last page：645, Aug. 2010, [Reviewed]
  Wound healing consists of sequential steps of tissue repair, and cell migration is particularly important. In order to analyze the potential function of growth arrest and DNA damage inducible protein 34 (GADD34) in tissue repair, we performed in vitro and in vivo wound healing experiments. In an in vitro scratch assay, GADD34 knockout (KO) mouse embryonic fibroblasts (MEFs) had higher migration rates than did wild type (WT) MEFs. Furthermore, the rate of wound closure was faster in GADD34 KO MEFs than in WT MEFs. Using in vivo punch biopsy assays, GADD34 KO mice had accelerated wound healing compared to WT mice. WT mice expressed higher amounts of myosin IIA in migrating macrophages and myofibroblasts than did GADD34 KO mice. These results indicate that GADD34 negatively regulates cell migration in wound healing via expression of myosin IIA.
  SPRINGER, English, Scientific journal
  DOI：https://doi.org/10.1007/s11248-009-9347-z
  DOI ID：10.1007/s11248-009-9347-z, ISSN：0962-8819, PubMed ID：20625881, Web of Science ID：WOS:000279844300011
• The wound repair is control by monocyte linage cells　　　　　　　　　　　　　　　
  Naomi Nishio; Sachiko Ito; Yayoi Okawa; Ken-Ichi Isobe
  2010 International Symposium on Micro-NanoMechatronics and Human Science: From Micro and Nano Scale Systems to Robotics and Mechatronics Systems, MHS 2010, Micro-Nano GCOE 2010, Bio-Manipulation 2010, First page：278, Last page：282, 2010, [Reviewed]
  One of the most important clinical problems in caring for elderly patients is treatment of pressure ulcers. One component of normal wound healing is the generation of an inflammatory reaction, which is characterized by the sequential infiltration of neutrophils, macrophages and lymphocytes. In aged C57BL/6 mice, wound healing is relatively inefficient. We examined the effects of monocytes lineage cells for wound healing. First, we i.v.transplanted bone marrow cells from GFP mice (C57BL/6 background) to C57BL/6 mice. We found that the rate of wound repair was enhanced by the bone marrow transplantation. By Immunohistochemistry, we can detect GFP positive cells in wound site. By FACS analysis Gr-1+ (neutrophils) and CD11b+ cells (macrophages) are observed in wound tissues. These results indicate that macro phages and/or neutrophils have the capacity to repair wounded tissues. We compare the capacity to repair wound by aging. Although old bone marrow cells also enhanced wound repair by bone marrow transplantation, the rate of wound repair old bone marrow was lower than the transplantation of young bone marrow. These results indicate that wound healing is controlled by monocytes lineage cells including neutrophils and macro phages. These monocytes lineage cells of aged mice have lower capacity to repair wound than those of young mice. ©2010 IEEE.
  English, International conference proceedings
  DOI：https://doi.org/10.1109/MHS.2010.5669544
  DOI ID：10.1109/MHS.2010.5669544, SCOPUS ID：78751481881
• Antibodies to wounded tissue enhance cutaneous wound healing　　　　　　　　　　　　　　　
  Naomi Nishio; Sachiko Ito; Haruhiko Suzuki; Ken-Ichi Isobe
  IMMUNOLOGY, Volume：128, Number：3, First page：369, Last page：380, Nov. 2009, [Reviewed]
  The wound repair process is a highly ordered sequence of events that encompasses haemostasis, inflammatory cell infiltration, tissue regrowth and remodelling. Wound healing follows tissue destruction so we hypothesized that antibodies might bind to wounded tissues, which would facilitate the engulfment of damaged tissues by macrophages. Here, we show that B cells, which produce antibodies to damaged tissues, are engaged in the process of wound healing. Splenectomy delayed wound healing, and transfer of spleen cells into splenectomized mice recovered the delay in wound healing. Furthermore, wound healing in splenectomized nude mice was also delayed. Transfer of enriched B220(+) cells by magnetic beads accelerated wound healing in splenectomized mice. We detected immunoglobulin G1 (IgG1) binding to wounded tissues by using fluorescein isothiocyanate-labelled anti-IgG1 6-24 hr after wounding. Splenectomy reduced the amount of IgG1 binding to wounded tissues. Immunoblotting studies revealed several bands, which were reduced by splenectomy. Using immunoprecipitation with anti-IgG bound to protein G we found that the intensity of several bands was lower in the serum from splenectomized mice than in that from sham-operated mice. These bands were matched to myosin IIA, carbamoyl-phosphate synthase, argininosuccinate synthase, actin and alpha-actinin-4 by liquid chromatography tandem mass spectrometry analysis.
  WILEY-BLACKWELL PUBLISHING, INC, English, Scientific journal
  DOI：https://doi.org/10.1111/j.1365-2567.2009.03119.x
  DOI ID：10.1111/j.1365-2567.2009.03119.x, ISSN：0019-2805, PubMed ID：20067537, Web of Science ID：WOS:000270588300007
• [Age-related decline of immune function and age-related diseases].　　　　　　　　　　　　　　　
  Ken-ichi Isobe; Naomi Nishio; Sachiko Ito
  Nihon rinsho. Japanese journal of clinical medicine, Volume：67, Number：7, First page：1327, Last page：31, Jul. 2009, [Reviewed], ［Domestic magazine］
  Effects of aging on immune system are widespread. The development of T and B cells declines with age. The functions of matured T and B cell also decline with age. Consequently, infections present major clinical problems for elderly patients. Many of age-related diseases are related to innate immunity. For example, the pathogenesis of atherosclerosis and Alzheimer disease are related to macrophages (microglia). The Ox-LDL or A-beta induces macrophages or microglia to produce inflammatory cytokines, chemokines and matrix metalloproteinases. Recently neutrophils have been shown to be an important immune cells in atherosclerosis. Neutrophils secrete inflammatory cytokines. In wound healing neutrophils also work as first important immune cells, which affect age-related decline of wound repair.
  Japanese, Scientific journal
  ISSN：0047-1852, PubMed ID：19591280
• Injection of bleomycin in newborn mice induces autoimmune sialitis that is transferred by CD4 T cells　　　　　　　　　　　　　　　
  Hideaki Ishikawa; Sachiko Ito; Naomi Nishio; Yukio Yuzawa; Sei-ichi Matsuo; Ken-ichi Isobe
  IMMUNOLOGY AND CELL BIOLOGY, Volume：87, Number：4, First page：351, Last page：358, May 2009, [Reviewed]
  Bleomycin (BLM) induces cellular apoptosis or necrosis by producing reactive oxygen species, and has been used to induce scleroderma in adult mice. We wondered whether BLM induces the same pathological phenotype in newborn mice as in adult mice. BLM was subcutaneously administrated to newborn BALB/c mice. At 1 month of age, BLM-treated mice showed severe destruction of salivary glands with enlargement of nearby lymph nodes. These nodes contained CD4(+) T cells and B220(+) cells with high expression of MHC class II molecules. In addition, autoantibodies were detected by HEp-2 staining and western blotting. The cell transfer experiments were performed to evaluate the role of autoimmune phenomena in these pathological changes. Following the transfer of enriched CD4(+) T cells to 1-month-old BALB/c nude mice, the salivary glands were severely damaged with CD4(+) T cell and B220(+) cells infiltrations. The number of T-cell antigen receptor V beta 8.3(+) CD4(+) T cells was significantly increased in BLM-treated murine spleen. These findings will provide new insights into the causal factors of environment in autoimmunity and the relationship between autoreactive CD4(+) T cells and autoantibodies. Immunology and Cell Biology ( 2009) 87, 351-358; doi:10.1038/icb.2009.1; published online 10 February 2009
  NATURE PUBLISHING GROUP, English, Scientific journal
  DOI：https://doi.org/10.1038/icb.2009.1
  DOI ID：10.1038/icb.2009.1, ISSN：0818-9641, PubMed ID：19204734, Web of Science ID：WOS:000266208800013
• Thymic Involution Correlates with Severe Ulcerative Colitis Induced by Oral Administration of Dextran Sulphate Sodium in C57BL/6 Mice but not in BALB/c Mice　　　　　　　　　　　　　　　
  Shin Sasaki; Yoshiyuki Ishida; Naomi Nishio; Sachiko Ito; Ken-ichi Isobe
  INFLAMMATION, Volume：31, Number：5, First page：319, Last page：328, Oct. 2008, [Reviewed]
  There is accumulating evidence to support the interactions between psychological stress and inflammatory bowel disease (IBD). In order to elucidate the relationship between psycoimmunological stress and IBD, we examined the alteration of immune system during the disease course of experimental Ulcerative colitis(UC)-model induced by dextran sulfate sodium (DSS). When C57BL/6 mice were treated with 4.5% DSS, they developed progressive weight loss. In contrast, the same treatment applied to BALB/c mice led to a small weight loss from which they soon recovered. Surprisingly, we found significant involution of the thymus and a reduction in the number of double positive thymocytes in DSS-treated C57BL/6 mice but not in DSS-treated BALB/c mice. Double negative thymocytes, especially DN1 (CD25-CD44+) and DN2 (CD25+CD44+) thymocytes, were relatively upregulated. The weights of spleens were slightly increased in both C57BL/6 and BALB/c mice following oral administration of DSS. In C57BL/6 spleens, both CD4 and CD8 single positive T cells gradually decreased (day 3), then recovered (day 14) after treatment. Because oral administration causes starvation, we examined the effects of starvation on the thymus and spleen. Although involution of thymus was observed both in starvation and DSS-treatment, the weight of spleen was reduced only in starvation. Also, the population changes in thymocytes in starvation was different from DSS-treatment. The administration of the steroid inhibitor RU486 partially reversed the thymic involution in C57BL/6 mice, thus DSS-treated UC might induce psycoimmunological changes partly through hypothalamic-pituitary-adrenal axis.
  SPRINGER/PLENUM PUBLISHERS, English, Scientific journal
  DOI：https://doi.org/10.1007/s10753-008-9081-3
  DOI ID：10.1007/s10753-008-9081-3, ISSN：0360-3997, PubMed ID：18696222, Web of Science ID：WOS:000260059800005
• Neutrophil depletion delays wound repair in aged mice　　　　　　　　　　　　　　　
  Naomi Nishio; Yayoi Okawa; Hidetoshi Sakurai; Ken-ichi Isobe
  AGE, Volume：30, Number：1, First page：11, Last page：19, Mar. 2008, [Reviewed]
  One of the most important clinical problems in caring for elderly patients is treatment of pressure ulcers. One component of normal wound healing is the generation of an inflammatory reaction, which is characterized by the sequential infiltration of neutrophils, macrophages and lymphocytes. Neutrophils migrate early in the wound healing process. In aged C57BL/6 mice, wound healing is relatively inefficient. We examined the effects of neutrophil numbers on wound healing in both young and aged mice. We found that the depletion of neutrophils by anti-Gr-1 antibody dramatically delayed wound healing in aged mice. The depletion of neutrophils in young mice had less effect on the kinetics of wound healing. Intravenous G-CSF injection increased the migration of neutrophils to the wound site. While the rate of wound repair did not change significantly in young mice following G-CSF injection, it increased significantly in old mice.
  SPRINGER, English, Scientific journal
  DOI：https://doi.org/10.1007/s11357-007-9043-y
  DOI ID：10.1007/s11357-007-9043-y, ISSN：0161-9152, PubMed ID：19424869, Web of Science ID：WOS:000255964700002
• Efficient induction of osteogenic and chondrogenic progenitors and myogenic progenitors from mouse ES cells in chemically defined medium　　　　　　　　　　　　　　　
  Hidetoshi Sakurai; Yuta Inami; Naomi Nishio; Tohru Yoshikai; Ken-Ichi Isobe
  2008 International Symposium on Micro-NanoMechatronics and Human Science, MHS 2008, First page：487, Last page：490, 2008, [Reviewed]
  Embryonic stem (ES) cells and induced pluripotent stem (iPS) cells represent a renewable cellular resources for regenerative therapies. Here, we try to differentiate ES cells to paraxial mesodermal progenitors cells (PMPs), which give rise to osteogenic and chondrogenic progenitors and myogenic progenitors without fetal calf serum. Here, we demonstrate the methods for inducing paraxial mesodermal progenitors efficiently by treatment of BMP4 using PDGFR-a and ECD as markers for purifying them. The BMP4 induced PMPs exhibit fine potentials of differentiation to osto- and chondro-genic cells. Furthermore, early extraction of BMP4 and additional LiCl treatment promote the differentiation of myogenic progenitor cells. Further addition of IGF-1, bFGF and HGF induced the differentiation of PMPs to myocytes in vitro. © 2008 IEEE.
  English, International conference proceedings
  DOI：https://doi.org/10.1109/MHS.2008.4752501
  DOI ID：10.1109/MHS.2008.4752501, SCOPUS ID：62449169784
• Paraxial mesodermal progenitors derived from mouse embryonic stem cells contribute to muscle regeneration via differentiation into muscle satellite cells　　　　　　　　　　　　　　　
  Hidetoshi Sakurai; Yayoi Okawa; Yuta Inami; Naomi Nishio; Ken-ichi Isobea
  STEM CELLS, Volume：26, Number：7, First page：1865, Last page：1873, 2008, [Reviewed]
  Pluripotent embryonic stem (ES) cells hold great potential for cell-based therapies. Although several recent studies have reported the potential of ES cell-derived progenitors for skeletal muscle regeneration, how the cells contribute to reconstitution of the damaged myofibers has remained elusive. Here, we demonstrated the process of injured muscle regeneration by the engraftment of ES cell-derived mesodermal progenitors. Mesodermal progenitor cells were induced by a conventional differentiation system and isolated by flow cytometer of platelet-derived growth factor receptor-alpha (PDGFR-alpha), a marker of paraxial mesoderm, and vascular endothelial growth factor receptor-2 (VEGFR-2), a marker of lateral mesoderm. The PDGFR-alpha(+) population that represented the paraxial mesodermal character demonstrated significant engraftment when transplanted into the injured muscle of immunodeficient mouse. Moreover, the PDGFR-alpha(+) population could differentiate into the muscle satellite cells that were the stem cells of adult muscle and characterized by the expression of Pax7 and CD34. These ES cell-derived satellite cells could form functional mature myofibers in vitro and generate myofibers fused with the damaged host myofibers in vivo. On the other hand, the PDGFR-alpha(-)VEGFR-2(+) population that showed lateral mesodermal character exhibited restricted potential to differentiate into the satellite cells in injured muscle. Our results show the potential of ES cell-derived paraxial mesodermal progenitor cells to generate functional muscle stem cells in vivo without inducing or suppressing gene manipulation. This knowledge could be used to form the foundation of the development of stem cell therapies to repair diseased and damaged muscles.
  ALPHAMED PRESS, English, Scientific journal
  DOI：https://doi.org/10.1634/stemcells.2008-0173
  DOI ID：10.1634/stemcells.2008-0173, ISSN：1066-5099, PubMed ID：18450822, Web of Science ID：WOS:000258004400022
• Differential effects of physical and psychological stressors on immune functions of rats　　　　　　　　　　　　　　　
  K Oishi; N Nishio; K Konishi; M Shimokawa; T Okuda; T Kuriyama; K Machida
  STRESS-THE INTERNATIONAL JOURNAL ON THE BIOLOGY OF STRESS, Volume：6, Number：1, First page：33, Last page：40, Mar. 2003, [Reviewed]
  To study the effects of different types or durations of stressors on immune functions, male Fischer rats were exposed to chronic physical (electric foot shock) or psychological (non-foot shock) stress induced in the communication box. Superoxide production by alveolar macrophages (AMs), mitogen-induced splenic lymphocyte proliferation, and splenic natural killer (NK) cell cytolysis were examined in vitro. Repeated exposure to physical stress suppressed superoxide production by AMs (-58%, p < 0.05 for opsonized zymosan (OZ) and -51%, p < 0.05 for phorbol 12-myristate 13-acetate (PMA)), although psychological stress suppressed superoxide production after 24 h of repeated exposures (-40%, p < 0.05 for OZ and -47%, p < 0.05 for PMA). Acute suppression of the blastic response of splenic lymphocytes was only found in the physical stress group (p < 0.05), although the chronic effects were only found in the psychological stress group (p < 0.05). NK cell activity was suppressed immediately after the acute physical stress (-30%, p < 0.05), but no effects were found in the psychological stress group. These results underline the importance of distinguishing between physical versus psychological stressors when examining the effects of stress on immune functions.
  TAYLOR & FRANCIS LTD, English, Scientific journal
  DOI：https://doi.org/10.1080/1025389031000101330
  DOI ID：10.1080/1025389031000101330, ISSN：1025-3890, PubMed ID：12637205, Web of Science ID：WOS:000184580400006
• SST-2 tumor inoculation is a useful model for studying the anti-tumor immune response in SHR rats　　　　　　　　　　　　　　　
  Naomi Nishio; Katsutaka Oishi; Kazuhiko Machida
  Environmental Health and Preventive Medicine, Volume：8, Number：1, First page：1, Last page：5, 2003, [Reviewed]
  Objective: The purpose of the present study was to investigate the relation between the dose of tumor cell inoculation (especially the doses less than minimum required to evoke tumor growth) and the anti-tumor immune system, particularly lymphoblast formation and cytotoxic activity of lymphcytes. Method: We inoculated rats with various doses of SST-2 tumor cells and examined natural killer (NK) cell activity and lymphoblast formation in vitro. Result: The results showed that the cytotoxicities against SST-2 cells and lymphoblast formation of lymphocytes were enhanced by small dose inoculation of tumor cells that could not induce tumor growth. Conclusion: It was suggested that was lymphocutes play an important role as an anti-tumor immune system at small doses of tumor inoculation, which appears to reflect an early stage of tumor growth in vivo. It was also suggested that SST-2 tumor inoculation might be a useful model for studying the anti-tumor immune response in SHR rats.
  Japanese Society for Hygiene, English, Scientific journal
  DOI：https://doi.org/10.1007/BF02897936
  DOI ID：10.1007/BF02897936, ISSN：1342-078X, PubMed ID：21432108, SCOPUS ID：0037358027
• [The effect of chronic stress on neutrophil function in tumor-inoculated rats].　　　　　　　　　　　　　　　
  Nishio N; Fujioka Y; Midorikawa T; Ishizaki K; Machida K
  Nihon eiseigaku zasshi. Japanese journal of hygiene, Volume：57, Number：4, First page：655, Last page：660, Jan. 2003, [Reviewed]
  Objectives: In order to clarify the relation between stress load and inoculated tumor growth in conjunction with neutrophil functions, several kinds of stress such as physical (Ph) and psychological (Ps) stress were loaded on rats either SST-2 tumor cell inoculated or control, and the functions of their peripheral neutrophils were determined.
  Methods: A communication box was used for stress load on rats. SST-2 cells were inoculated in to rats in the tumor-inoculation groups. Two weeks after inoculation, the tumors were removed from the backs of the rats and weighed. The functions of neutrophils in the peripheral blood collected from the tail vein, were determined by the NBT deoxidization method.
  Results: Tumor growth was enhanced when rats were loaded with either Ph or Ps stresses, but was inhibited when tumor cells were inoculated following either Ph or Ps stresses. These results show that chronological differences of loaded stresses influence immunological functions differently. The O₂^- production from the neutrophils stimulated by NBT-treated Staphilococcus aureus was suppressed in tumor-inoculated Ph and Ps groups, more markedly in the tumor-inoculated Ph group. It is logically relevant that the size of tumors increased in these groups, predominantly in the tumor-inoculated Ph group. On the other hand, O₂^- production from the neutrophils was enhanced and tumor growth decreased in tumor-inoculated animals following either Ph or Ps stresses.
  Conclusions: Our experiments, it revealed that the function of neutrophils is strongly enhanced by stress load and O₂^- production is inhibited by the tumor inoculation as shown in stimulation tests. Therefore, our findings suggested that neutrophils may participate in the inhibition of tumor growth.
  The Japanese Society for Hygiene, Japanese
  DOI：https://doi.org/10.1265/jjh.57.655
  DOI ID：10.1265/jjh.57.655, ISSN：0021-5082, CiNii Articles ID：130000997718, CiNii Books ID：AN00185923, PubMed ID：12638170

• Loss of GADD34 induces early age- dependent deviation to the myeloid lineage　　　　　　　　　　　　　　　
  Naomi Nishio; Sachiko Ito; Ken-ichi Isobe
  IMMUNOLOGY AND CELL BIOLOGY, Volume：92, Number：2, First page：170, Last page：180, Feb. 2014
  Hematopoietic stem cells (HSCs) generate all known hematopoietic lineages and are capable of self-renewal. Upon aging, myeloid-biased HSCs are maintained, whereas lymphoid-biased HSCs are lost. GADD34 protein is expressed in myeloid-lineage cells and has been cloned from them. However, the function of GADD34 in the myeloid lineage has not yet been elucidated. Here, we show that early age-dependent deviation to the myeloid lineage occurs in GADD34-deficient mice. Early increases of GR-1(int)CD11b(+) and GR-1(high)CD11b(+) neutrophils were observed in the spleen, bone marrow (BM) and blood of GADD34-deficient mice. We found that BM Lin(-) c-Kit(+) Sca1(+) and Lin(-) c-Kit(+) Sca1(-) cells expressed GADD34 protein without stimulation and increased GADD34 expression following intravenous injection of Staphylococcus aureus (S. aureus). These cell populations were high in GADD34-deficient BM and were increased by the injection of S. aureus. Because of the increase in granulocyte colony-stimulating factor (G-CSF) induced by S. aureus injection, we examined the signaling pathway from the G-CSF receptor (G-CSFR). We found that phosphorylation of signal transducer and activator of transcription factor 3 was highly increased in GADD34-deficient Lin(-) BM cells by the stimulation of G-CSF. These results indicate that GADD34 binds to Lyn and inhibit G-CSFR signaling. We show here that GADD34 works to inhibit the proliferation and differentiation of HSCs or myeloid precursor cells and maintains homeostatic differentiation of neutrophil-lineage cells to avoid early immunological senescence.
  NATURE PUBLISHING GROUP, English
  DOI：https://doi.org/10.1038/icb.2013.78
  DOI ID：10.1038/icb.2013.78, ISSN：0818-9641, eISSN：1440-1711, Web of Science ID：WOS:000330999500012
• Transplantation of Induced Neural Crest Cells from Spheroidal Dermal Stem Cells Improves Diabetic Polyneuropathy in Mice　　　　　　　　　　　　　　　
  Masaki Kondo; Hideki Kamiya; Tatsuhito Himeno; Tetsuji Okawa; Jiro Kato; Atsushi Hujiya; Yoji Hamada; Keiko Naruse; Naomi Nishio; Sachiko Ito; Yutaka Oiso; Ken-Ichi Isobe; Jiro Nakamura
  DIABETES, Volume：62, First page：A146, Last page：A146, Jul. 2013
  AMER DIABETES ASSOC, English, Summary international conference
  ISSN：0012-1797, eISSN：1939-327X, Web of Science ID：WOS:000209473600571
• Transplantation of Neural Progenitor-Like Cells Derived from Spheroidal Dermal Cells Improves Diabetic Polyneuropathy in Mice　　　　　　　　　　　　　　　
  Masaki Kondo; Hideki Kamiya; Tatsuhito Himeno; Jiro Kato; Tetsuji Okawa; Atsushi Fujiya; Keiko Naruse; Yoji Hamada; Naomi Nishio; Sachiko Ito; Yutaka Oiso; Ken-Ichi Isobe; Jiro Nakamura
  DIABETES, Volume：61, First page：A20, Last page：A20, Jun. 2012
  AMER DIABETES ASSOC, English, Summary international conference
  ISSN：0012-1797, eISSN：1939-327X, Web of Science ID：WOS:000209842900071
• Analysis of beta-Amyloid Peptide -Binding Proteins in Microglial Cells　　　　　　　　　　　　　　　
  Sachiko Ito; Naomi Nishio; Ken-chi Isobe
  The Open Geriatric Medicine Journal, Volume：5, First page：1-6, 2012, [Reviewed]
  English, Report scientific journal
• The change of neutrophil function on tumor inoculated rats　　　　　　　　　　　　　　　
  NISHIO Naomi
  Volume：56, Number：1, First page：235, Last page：235, 01 Apr. 2001
  Japanese
  ISSN：0021-5082, CiNii Articles ID：10012355772, CiNii Books ID：AN00185923
• The effects of physical and pshychological stress on rat injected tumor cells : (2) Nonspecific defense function　　　　　　　　　　　　　　　
  NISHIO Naomi
  Volume：54, Number：1, First page：384, Last page：384, 15 Apr. 1999
  Japanese
  ISSN：0021-5082, CiNii Articles ID：10013389224, CiNii Books ID：AN00185923
• The effects of physical and pshychological stress on rat injected tumor cells : (1) Biological function　　　　　　　　　　　　　　　
  FUJIOKA Yosei
  Volume：54, Number：1, First page：383, Last page：383, 15 Apr. 1999
  Japanese
  ISSN：0021-5082, CiNii Articles ID：10013389223, CiNii Books ID：AN00185923

• Apr. 2018 - Present
  微生物学(免疫学を含む)
• Apr. 2018 - Present
  生理学
• Apr. 2018 - Present
  解剖生理学
• Apr. 2018 - Present
  臨床実習
• 2018 - Present
  栄養学(食品学を含む)
• Apr. 2019 - Mar. 2022
  薬理概論
• Oct. 2018 - Mar. 2022
  physiology, Tokyo Gakugei University
• Oct. 2019 - Mar. 2020
  栄養学
• Apr. 2018
  解剖生理学実習

• THE JAPANESE SOCIETY FOR HYGIENE

• 「チーム学校」による心身緊急時等対応体制構築のための教育プログラム開発と情報支援　　　　　　　　　　　　　　　
  01 Apr. 2022 - 31 Mar. 2027
  Grant amount(Total)：16510000, Direct funding：12700000, Indirect funding：3810000
  Grant number：22H00961
• GADD34による老化関連疾患の制御メカニズムの解析　　　　　　　　　　　　　　　
  01 Apr. 2019 - 31 Mar. 2023
  Grant amount(Total)：4290000, Direct funding：3300000, Indirect funding：990000
  Grant number：19K11687
• Stress responsive GADD34 and metabolic syndrome　　　　　　　　　　　　　　　
  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (B), 01 Apr. 2016 - 31 Mar. 2019
  isobe ken-ichi, Nagoya Women's University
  Grant amount(Total)：17290000, Direct funding：13300000, Indirect funding：3990000
  metabolic syndrome represent serious in Japan. We found that aged GADD34 -deficient mice showed fatty liver.We examined the effects of GADD34 on natural life span by using GADD34-deficient mice. We observed that with age GADD34-deficient mice become obese, developing fatty liver followed by liver cirrhosis, hepatocellular carcinoma and insulin resistance. During these studies, we found that nutrients in the foods were important to induce fatty liver. Feeding of sugar, glucose or fructose only diet induced strong nonalcoholic fatty liver disease only in 2 weeks.. High fats (60% fat) without choline diet also induced fatty liver in 2 weeks. For more than one year feeding of adult C57BL/6 female mice egg-only diet did not induce fatty liver. Adding boiled eggs or law egg yolk to sugar, glucose or fructose or only diet or colline deficient diet strongly suppressed fatty liver.
  Grant number：16H05280
• Age-related diseases analyzed by ER stresses and immune functions using murine models including iPSCs　　　　　　　　　　　　　　　
  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (B), 01 Apr. 2013 - 31 Mar. 2017
  isobe ken-ichi
  Grant amount(Total)：18330000, Direct funding：14100000, Indirect funding：4230000
  We intended to analyze age-related disease from the points of ER stresses and immune responses. Acrolein, which is abundant in cigarette smoke, and cooking emission,
  plays a major role in COPD . Acrolein induced the expression of GADD34 and acute inflammation in airways, which followed COPD. Acrolein-induced phosphorylation of eIF2α in GADD34-knockout epithelial cells by shRNA protected cell death by reducing misfolded protein-caused oxidative stress. We examine the effects of GADD34 on natural life span by using GADD34-deficient mice. We found that with age GADD34-deficient mice become obese, developing fatty liver followed by liver cirrhosis, hepatocellular carcinoma, and insulin resistance. GADD34 suppresses insulin signaling in young mice. However, by aging macrophages in fat tissues work to induce insulin resistance. We succeeded to differentiate to various tissue cells from iPSCs established from aged mice, which may repair damaged tissues by age-related diseases.
  Grant number：25293166