Epigenetic regulation of proliferation and differentiation of hematopoietic stem cells

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 17390300
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Pediatrics
• Research Institution: Shinshu University
• Principal Investigator: KOIKE Kenichi Shinshu University, Department of Pediatrics, Professor (40143979)
• Co-Investigator(Kenkyū-buntansha): SHIOHARA Masaaki Shinshu University, Department of Pediatrics, Assistant Professor (00293514) ; NAKAZAWA Yozo Shinshu University, Department of Pediatrics, Senior Assistant Professor (60397312)
• Project Period (FY): 2005 – 2007
• Keywords: Stem cells / GM-CSF / p15 / methylation / histone acetylation / MO7F Cells / AML

Research Abstract

In the present study, to understand the process of GM-CSF-induced methylation in the CpG. island region of p15, we examined the methylation status of the gene in a human GM-CSF-dependent megakaryoblastic leukemic cell line, MO7_c, using bisulfite genomic sequencing. The process of p15 CpG island methylation induced by granulocyte-macrophage colony-stimulating factor (GM-CSF) was investigated, using MO7_c cells. The cells proliferating in response to GM-CSF plus fetal bovine serum (FBS) were almost fully methylated in the p15 CpG island. The withdrawal of both GM-CSF and IBS for 48 hours reduced the cell viability, and increased the frequency of alleles with completely or partially demethylated CpG sites by approximately 50%. Viable cells were responsible for this epigenetic change. The add-back of GM-CSF restored the methylation. Seventy-two hours withdrawal of GM-CSF plus FBS followed by 24 hours exposure to inhibitors for DNA methyltransferase (DNMT) and histone deacctylase (HDAC) ca used the demethylation of nearly all CpG sites in the p15 CpG island on every allele sequenced. When GM-CSF was re-added after the 96-hour treatment, the cells exhibited p15 transcriptional silencing via the methylation. The initial methylation event encompassed the entire CpG island. No new methylated alleles appeared in the coexistence of the DNMT and HDAC inhibitors. Taken together, GM-CSF may be able to induce de novo methylation of the p15 gene, using 11DAC(s) as well as DNMT(s).
Histone modifications regulating expression of the genes which encode CDK inhibitors in normal peripheral blood/bone marrow cells and acute myeloblastic leukemia (AML) remain unclear. We attempted, for the first time, to elucidate histone modifications around the CpG island region of the p15 gene in AML cells, using the chromatin immunoprecipitation (ChIP) assay and methylation-specific PCR with bisulfite genomic sequencing. Seven of 11 patients with AML had allele(s) in which more than half of 27 CpG sites in the p15 gene were methylated. The p15 CpG island region was surrounded with both the acetylated histone H3 (A_cH3) and dimethylated histone H3-lysine 9 (M_eH3K9) in bone marrow cells of AML patients, whereas with A_cH3 alone in normal marrow cells. The p 15 CpG islands of DNA immunoprecipitated with anti-A_cH3 antibody and anti-M_eH3K9 antibody were not always unmethylated and methylated, respectively, in the patients. These results suggest perturbed modifications of histone H3 around the p15 CpG island region in AML.

Research Products

• [Journal Article] Recent advances in the pathogenesis and management of juvenile mvelomonocytic leukemia. (2008)
  • Author(s): Koike K. Matsuda K.
  • Journal Title: Br J Haematol. 141(In press) Pages: 567-75
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Anakinra improved sensory deafness in a Japanese patient with Muckle-Wells syndrome by Possibly inhibiting cryopyrininflammasome. (2008)
  • Author(s): Yamazaki T, Masumoto J, Agematsu K, Sawai N, Kobayashi S, Shigemura T, Yasui K, Koike K.
  • Journal Title: Arthritis Rheum 2007 58(In press) Pages: 864-8
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Neurodegenerative central nervous system disease as late sequelae of Langerhans cell histiocytosis. (2008)
  • Author(s): Imashuku S, Koike K, 他12名, 4番目.
  • Journal Title: Haematologica, 93(in press.) Pages: 615-8
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Acquirement of loss of the wild-type NRAS locus with aggres sive disease progression in a patient with juvenile myelomonocytic leukemia and a heterozygous NRAS mutation. (2007)
  • Author(s): Matsuda K, Nakazawa Y. Sakashita K, Shiohara M, Yamauchi K, Koike K.
  • Journal Title: Haematologica. 92 Pages: 1576-1578
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Treatment of neurodegenerative CNS disease in langerhans cell histiocytosis with a combination of intravenous immunoglobulin and chemotherapy. Pediatr (2007)
  • Author(s): Imashuku S, Okazaki NA, Nakayama M, Fujita N, Fukuyama T, Koike K, Minato T, Kobayashi R, Morimoto A
  • Journal Title: Blood Cancer 50 Pages: 308-11
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Spontaneous improvement of hematologic abnormalities in patients having juvenile myelomonocytic leukemia with specific RAS mutations (2007)
  • Author(s): Matsuda K, Shimada A, Yoshida N. Ogawa A, Watanabe A, Yajima S, Iizuka S, Koike K, Yanai F, Kawasaki K, Yanagimachi M, Kikuchi A, Ohtsuka Y, Hidaka E, Yamauchi K, Tanaka M, Yanagisawa R, Nakazawa Y, Shiohara M, Manabe A. Kojima S, Koike K.
  • Journal Title: blood, 109 Pages: 5477-80
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] A complex karyotype. including a three-way translocation generating a NUP98-HOXD13 transcript. in an infant with acute myeloid leukemia. (2007)
  • Author(s): Hidaka E, Tanaka M, Matsuda K, Ishikawa-Matsumura M, Yamauchi K, Sano K, Honda T, Wakui K, Yanagisawa R, Nakazawa Y, Sakashita K, Shiohara M, Ishii E, Koike K.
  • Journal Title: Cancer Genet Cytogenet. 176 Pages: 137-43
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Late-onset cases of familial hemophagocytic lymphohistiocytosis with missense perforin gene mutations. (2007)
  • Author(s): Ueda I, Kurokawa Y, Koike K, Ito S, Sakata A, Matsumora T, Fukushima T, Morimoto A, Ishii E, Imashuku S.
  • Journal Title: Am J Hematol 82 Pages: 427-32
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Analysis of histone modification around the CpG island region of the pl5 gene in acute myeloblastic leukemia (2007)
  • Author(s): Ogawa M, Sakashita K, Zhao XY, Hayakawa A, Kubota T, Koike K
  • Journal Title: Leukemia Res 31 Pages: 611-21
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] A possible role for the production of multiple HLA antibodies in fatal platelet transfusion refractoriness after peripheral blood stem cell transplantation from the mother in a patient with relapsed leukemia (2007)
  • Author(s): Nakazawa Y, Saito S, Hasegawa Y, Yanagisawa Y, Sakashita K, Kamijo T, Miyazaki T, Sato S, Ikeda H, Ikebuchi K, Koike K
  • Journal Title: Transfusio 47 Pages: 326-34
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Chromosomal change during 6-mercaptopurine(6-MP)therapy in juvenile myelomonocytic leukemia: the growth of a 6-MP-refractory clone that already exists at onset. (2006)
  • Author(s): Matsuda K, Matsuzaki S, Miki J, Hidaka E, Yanagisawa R, Nakazawa Y, Sakashita K, Kamijo T, Asami K, Sano K, Koike K
  • Journal Title: Leukemia 20 Pages: 485-490
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Allogeneic haematopoietic cell transplantation from alternative donors with a conditioning regimen of low-dose irradiation. fludarabine and cyclophosphamide in Fanconi anaemia. (2006)
  • Author(s): Yabe H. Inoue H. Matsumoto M. Hamanoue S. Koike T. Ishiguro H. Koike H. Suzuki K, Kato S. Kojima S. Tsuchida M. Mori T. Adachi S. Tsuji K. Koike K. Morimoto A. Sako M. Yabe M
  • Journal Title: Br J Haematol. 134 Pages: 208-12
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Genetic subtypes of familial hemophagocytic lymphohistiocytosis: correlations with clinical features and cytotoxic T lymphocyte/natural killer cell functions (2005)
  • Author(s): Ishii E, Ueda I, Shirakawa R, Yamamoto K, Morimoto A, Imayoshi M, Ogata Y, Zaitsu M, Sako M, Koike K, Sakata A, Takada H, Hara T, Imashuku S, Sasazuki T, Yasukawa M.
  • Journal Title: Blood 105 Pages: 3442-8
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Granulocyte-macrophage colony-stimulating factor induces de novo methylation of the p15 CpG island in hematopoietic cells (2005)
  • Author(s): Zhao XY, Sakashita K, Kamijo T, Hidaka E, Sugane, K, Kubota T, Koike K
  • Journal Title: Cytokine, 31 Pages: 203-212
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Development of two cytogenetically abnormal clones from multipotential hematopoietic stem cells in a patient with juvenile myelomonocytic leukemia. (2005)
  • Author(s): Matsuzaki S. Matsuda K, Miki J, Nakazawa Y, Sakashita K, Kamijo T, Hidaka E, Koike K
  • Journal Title: Leukemia Res, 29 Pages: 1069-1072
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Methylation status of the ρ15 and ρ16 genes in paediatric myelodysplastic syndrome and juvenile myelomonocytic leukaemia (2005)
  • Author(s): Hasegawa D, Manabe A, Kubota T, Kawasaki H, Hirose I, Ohtsuka Y, Tsuruta T, Ebihara Y, Goto Y, Zhao XY. Sakashita K, Koike K, Isomura M, Kojima S, Hoshika A, Tsuji K, and Nakahata T
  • Journal Title: Br J Heamatol 128 Pages: 805-12
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Presentation] Recent advances in the pathogenesis and management of juvenile myelomonocytic leukemia (2008)
  • Author(s): Koike, K., Matsuda, K
  • Organizer: Br J Haematol
  • Year and Date: 20080000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Anakinra improved sensory deafness in a Japanese patient with Muckle-Wells syndrome by possibly inhibiting cryopyrin inflammasome (2008)
  • Author(s): Yamazaki, T., Masumoto, J., Agematsu, K., Sawai, N., Kobayashi, S., Shigemura, T., YasuiK, Koike, K
  • Organizer: Arthritis Rheum 2007
  • Year and Date: 20080000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Neurodegenerative central nervous system disease as late sequelae of Langerhans cell histiocytosis (2008)
  • Author(s): Imashuku, S., Koike, K
  • Organizer: Haematologica
  • Year and Date: 20080000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 腸管型ベーチェット病を併発しtrisomy 8とPTPN11遺伝子異常を認めた骨髄異形成症候群(RAEB-M6)の1例 (2007)
  • Author(s): 重村倫成、高山和生、齋藤章治、田中美幸、柳沢 龍、坂下一夫、山崎崇志、塩原正明、上松一永、小池健一、松田和之、樋口由美子
  • Organizer: 第49回日本小児血液学会
  • Place of Presentation: 仙台市
  • Year and Date: 20071214-15
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 高IgMを合併したSLEの一例 (2007)
  • Author(s): 山崎崇志、日高義彦、重村倫成、石田岳史、上松一永、江原孝史、小池健一
  • Organizer: 第29回日本小児腎不全学会総会
  • Place of Presentation: 松江市
  • Year and Date: 20070927-28
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] t(8;21)急性骨髄性白血病に対する化学療法および同種骨髄移植後の微少残存病変の解析 (2007)
  • Author(s): 田中美幸、柳沢 龍、中沢洋三、塩原正明、小池健一
  • Organizer: 第110回日本小児科学会学術集会
  • Place of Presentation: 京都市
  • Year and Date: 20070420-22
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Acquirement of loss of the wild-type NRASlocus with aggressive disease progression in a patient with juvenile myelomonocytic leukemia and a heterozygous NRAS mutation (2007)
  • Author(s): Matsuda, K., Nakazawa, Y., Sakashita, K., Shiohara, M., Yamauchi, K., Koike, K
  • Organizer: Pediatr Blood Cancer
  • Year and Date: 20070000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Spontaneous improvement of hematologic abnormalities in patients having juvenile myelomonocytic leukemia with specific RAS mutations (2007)
  • Author(s): Matsuda, K., Shimada, A., Yoshida, N., Ogawa, A., Watanabe, A., Yajima, S., Iizuka, S., Koike, K., Yanai, F., Kawasaki, K., Yanagimachi, M., Kikuchi, A., Ohtsuka, Y., Hidaka, E., Yamauchi, K., Tanaka, M., Yanagisawa, R., Nakazawa, Y., Shiohara, M., Manabe, A., Kojima, S., Koike, K
  • Organizer: Blood
  • Year and Date: 20070000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] A complex karyotype, including a three-way translocation generating a NUP98-HOXD13 transcript, in an infant with acute myeloid leukemia (2007)
  • Author(s): Hidaka, E., Tanaka, M., Matsuda, K., Ishilawa-Matsumura, M., Yamauchi, K., Sano, K., Honda, T., Wakui, K., Yanagisawa, R., Nakazawa, Y., Sakashita, K., Shiohara, M., Ishii, E., Koike, K
  • Organizer: Cancer Genet Cytogenet
  • Year and Date: 20070000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Late-onset cases of familial hemophagocytic lymphohistiocytosis with missense perforin gene mutations.Am J Hematol (2007)
  • Author(s): Ueda, I., Kurokawa, Y., Koike, K., Ito, S., Sakata, A., Matsumora, T., Fukushima, T., Morimoto, A., Ishii, E., Imashuku, S
  • Year and Date: 20070000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Analysis of histone modification around the CpG island region of the p 15 gene in acute myeloblastic leukemia (2007)
  • Author(s): Ogawa, M., Sakashita, K., Zhao, XY., Hayakawa, A., Kubota, T., Koike, K
  • Organizer: Leukemia Res
  • Year and Date: 20070000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] A possible role for the production of multiple HLA antibodies in fatal platelet transfusion refractoriness after peripheral blood stem cell transplantation from the mother in a patient with relapsed leukemia (2007)
  • Author(s): Nakazawa, Y., Saito, S., Hasegawa, Y., Yanagisawa, Y., Sakashita, K., Kamijo, T., Miyazaki, T., Sato, S., Ikeda, H., Ikebuchi, K., Koike, K
  • Organizer: Transfusion
  • Year and Date: 20070000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] 急性骨髄性白血病に対する同種骨髄移植後のAML1-ETOキメラ遺伝子による微少残存病変の解析 (2006)
  • Author(s): 田中美幸、柳沢龍、中沢洋三、塩原正明、小池健一、松田和之、鈴木剛史、川久保雅友
  • Organizer: 第68回日本血液学会・第48回日本臨床血液学会
  • Place of Presentation: 福岡市
  • Year and Date: 20061006-08
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 小児科領域におけるITの活用 (2006)
  • Author(s): 小池 健一
  • Organizer: 第109回日本小児科学会学術集会
  • Place of Presentation: 金沢市
  • Year and Date: 20060421-23
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 全身放射線照射量を減量した前処置による造血幹細胞移植の治療成績 (2006)
  • Author(s): 柳沢 龍、田中美幸、中沢洋三、塩原正明、上條岳彦、小池健一
  • Organizer: 第109回日本小児科学会学術集会
  • Place of Presentation: 金沢市
  • Year and Date: 20060421-23
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Chromosomal change during 6-mercaptopurine (6-MP) therapy in juvenile myelomonocytic leukemia : the growth of a 6-MP-refractory clone that already exists at onset (2006)
  • Author(s): Matsuda, K., Matsuzaki, S., Mild, J., Hidaka, E., Yanagisawa, R., Nakazawa, Y., Sakashita, K., Kamijo, T., Asami, K., Sano, K., Koike, K
  • Organizer: Leukemia
  • Year and Date: 20060000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Allogeneic haematopoietic cell transplantation from alternative donors with a conditioning regimen of low-dose irradiation, fludarabine and cyclophosphamide in Fanconi anaemia (2006)
  • Author(s): Yabe, H., Inoue, H., Matsumoto, M., Hamanoue, S., Koike, T., Ishiguro, H., Koike, H., Suzuki, K., Kato, S., Kojima, S., Tsuchida, M., Mod, T., Adachi, S., Tsuji, K., Koike, K., Morimoto, A., Sako, M., Yabe, M
  • Organizer: Br J Haematol
  • Year and Date: 20060000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Three-wayおよびfour-way translocationを示した乳児白血病の2例 (2005)
  • Author(s): 田中美幸, 柳沢龍, 中沢洋三, 塩原正明, 上條岳彦, 小池健一, 松田和之、鈴木剛史、日高恵以子
  • Organizer: 臨床血液学会
  • Place of Presentation: 横浜
  • Year and Date: 20050917-19
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] 若年性慢性骨髄性白血病におけるp15遺伝子のDNAメチル化とヒストン修飾 (2005)
  • Author(s): 小川美奈、坂下一夫、小池健一
  • Organizer: 第108回日本小児科学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 20050422-23
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Genetic subtypes of familial hemophagocytic lymphohistiocytosis : correlations with clinical features and cytotoxic T lymphocyte/natural killer cell functions (2005)
  • Author(s): Ishii, E., Ueda, I., Shirakawa, R., Yamamoto, K., Horiuchi, H., Ohga, S., Furuno, K., Morimoto, A., Imayoshi, M., Ogata, Y., Zaitsu, M., Sako, M., Koike, K., Sakata, A., Takada, H., Hara, T., Imashuku, S., Sasazuki, T., Yasukawa, M
  • Organizer: Blood
  • Year and Date: 20050000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Granulocyte-macrophage colony-stimulating factor induces de novo methylation of the p15 CpG island in hematopoietic cells (2005)
  • Author(s): Zhao, XY., Sakashita, K., Kamijo, T., Hidaka, E., Sugane, K., Kubota, T., Koike, K
  • Organizer: Cytokine
  • Year and Date: 20050000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Development of two cytogenetically abnormal clones from multipotential hematopoietic stem cells in a patient with juvenile myelomonocytic leukemia (2005)
  • Author(s): Matsuzaki, S., Matsuda, K., Miki, J., Nakazawa, Y., Sakashita, K., Kamijo, T., Hidaka, E., Koike, K
  • Organizer: Leukemia Res
  • Year and Date: 20050000
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Methylation status of the p15 and p16 genes in paediatric myelodysplastic syndrome and juvenile myelomonocytic leukaemia (2005)
  • Author(s): Hasegawa, D., Manabe, A., Kubota, T., Kawasaki, H., Hirose, I., Ohtsuka, Y., Tsuruta, T., Ebihara, Y., Goto, Y., Zhao, XY., Sakashita, K., Koike, K., Isomura, M., Kojima, S.,Hoshika, A., Tsuji, K., Nakahata, T
  • Organizer: Br J Heamatol
  • Year and Date: 20050000
  • Description: 「研究成果報告書概要(欧文)」より