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Detection method for quantifying global DNA methylation byfluorescence correlation spectroscopy

Research Project

Project/Area Number 23701087
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Tumor diagnosis
Research InstitutionTokyo Medical University

Principal Investigator

UMEZU Tomohiro  東京医科大学, 医学部, 助教 (40385547)

Project Period (FY) 2011 – 2012
Project Status Completed (Fiscal Year 2012)
Budget Amount *help
¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2011: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywordsゲノム / DNA メチル化 / 脱メチル化薬 / 1分子蛍光分析法 / MBD2 / メチル化DNA / DNA脱メチル化薬 / 一分子蛍光分析法 / 骨髄異形成症候群
Research Abstract

A method for quantifying global DNA methylation using fluorescence correlation spectroscopy (FCS) has been established. The single-molecule methylation assay (SMMA) is based on two methodologies. One methodology, FCS, estimates the translational diffusion coefficient of molecules in solution, whereas the other methodology uses the high affinity of methyl-CpG-binding domain protein 2 (MBD2) to bind specifically to methylated DNA. We studied the specific binding rates of fluorescence-labeled MBD2 and methylated DNA from biological samples using the automated FCS system. Using a standard curve with methylated control DNA, we developed the SMMA index to assess the global DNA methylation level of the biological samples. A marked decrease in the SMMA index was observed when human leukemia cell lines (U937 and K562) were cultured with DNA demethylating agents. Our findings clearly indicate the applicability of SMMA as a simple and rapid tool for quantifying global DNA methylation. SMMA may prove useful for genome-wide comparative methylation analyses of malignancies and as an indicator of the demethylation effects of epigenetic drugs.

Report

(3 results)
  • 2012 Annual Research Report   Final Research Report ( PDF )
  • 2011 Research-status Report
  • Research Products

    (19 results)

All 2012 2011 2010

All Journal Article (7 results) (of which Peer Reviewed: 7 results) Presentation (11 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] The C allele of JAK2 rs4495487 is an additional candidate locus that contributes to myeloproliferative neoplasm predisposition in the Japanese population.2012

    • Author(s)
      Ohyashiki JH, Yoneta M, Hisatomi H, Iwabuchi T, Umezu T, Ohyashiki K.
    • Journal Title

      BMC Med Genet.

      Volume: 13 Issue: 1 Pages: 6-6

    • DOI

      10.1186/1471-2350-13-6

    • Related Report
      2012 Annual Research Report 2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Downregulated plasma miR-92a levels have clinical impact on multiple myeloma and related disorders.2012

    • Author(s)
      Yoshizawa S, Ohyashiki JH, Ohyashiki M, Umezu T, Suzuki K, Inagaki A, Iida S, Ohyashiki K.
    • Journal Title

      Blood Cancer J.

      Volume: 2(1) Issue: 1 Pages: e53-e53

    • DOI

      10.1038/bcj.2011.51

    • Related Report
      2012 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Detection method for quantifying global DNA methylation by fluorescence correlation spectroscopy2011

    • Author(s)
      Umezu T, Ohyashiki K, Ohyashiki JH
    • Journal Title

      Anal Biochem

      Volume: 415(2) Issue: 2 Pages: 145-50

    • DOI

      10.1016/j.ab.2011.04.035

    • Related Report
      2012 Final Research Report 2011 Research-status Report
    • Peer Reviewed
  • [Journal Article] Clinical impact of down-regulated plasma miR-92a levels in non-Hodgkin's lymphoma2011

    • Author(s)
      Ohyashiki K, Umezu T, Yoshizawa S, Ito Y, Ohyashiki M, Kawashima H, Tanaka M, Kuroda M, Ohyashiki JH
    • Journal Title

      PLoS One

      Volume: 6(2) Issue: 2 Pages: 16408-16408

    • DOI

      10.1371/journal.pone.0016408

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Non-random chromosomal deletion clustering at 20q in Waldenstrom macroglobulinemia2011

    • Author(s)
      Kitahara T, Umezu T, Ando K, Kodama A, Ohyashiki JH, Ohyashiki K
    • Journal Title

      Hematology

      Volume: 16(3) Issue: 3 Pages: 139-42

    • DOI

      10.1179/102453311x12953015767338

    • Related Report
      2012 Final Research Report
    • Peer Reviewed
  • [Journal Article] Clinical impact of down-regulated plasma miR-92a levels in non-Hodgkin's lymphoma.2011

    • Author(s)
      Ohyashiki K, Umezu T, Yoshizawa S, Ito Y, Ohyashiki M, Kawashima H, Tanaka M, Kuroda M, Ohyashiki JH.
    • Journal Title

      PLoS One

      Volume: 6(2)

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Journal Article] Non-random chromosomal deletion clustering at 20q in Waldenström macroglobulinemia.2011

    • Author(s)
      Kitahara T, Umezu T, Ando K, Kodama A, Ohyashiki JH, Ohyashiki K.
    • Journal Title

      Hematology.

      Volume: 16(3) Pages: 139-142

    • Related Report
      2011 Research-status Report
    • Peer Reviewed
  • [Presentation] 骨髄異形成症候群におけるメチル化度測定による治療効果予測2012

    • Author(s)
      浅野倫代、小林千晶、廣田綾子、梅津知宏、大屋敷一馬、大屋敷純子
    • Organizer
      第170回東京医科大学医学会総会
    • Place of Presentation
      東京
    • Year and Date
      2012-11-17
    • Related Report
      2012 Final Research Report
  • [Presentation] Epigenetic aberration in leukocytes of patients with myelodysplastic syndrome2012

    • Author(s)
      浅野倫代、大屋敷純子、梅津知宏、藤本博昭、後藤守孝、赤羽大悟、北原俊彦、吉澤成一郎、大屋敷一馬
    • Organizer
      第74回日本血液学会学術集会
    • Place of Presentation
      京都
    • Related Report
      2012 Final Research Report
  • [Presentation] 骨髄異形成症候群 (MDS)のDNA メチル化度の評価;Single Molecular Methylation Assay (SMMA)法の有用性2012

    • Author(s)
      浅野倫代、大屋敷純子、梅津知宏、大槻和重、大屋敷一馬
    • Organizer
      第6回日本エピジェネティクス研究会年会
    • Place of Presentation
      東京
    • Related Report
      2012 Final Research Report
  • [Presentation] 骨髄異形成症候群 (MDS)のDNAメチル化度の評価;Single Molecular Methylation Assay (SMMA)法の有用性2012

    • Author(s)
      浅野倫代、大屋敷純子、梅津知宏、大槻和重、大屋敷一馬
    • Organizer
      第6回日本エピジェネティクス研究会年会
    • Place of Presentation
      東京
    • Related Report
      2012 Annual Research Report
  • [Presentation] Epigenetic aberration in leukocytes of patients with myelodysplastic syndrome.2012

    • Author(s)
      浅野倫代、大屋敷純子、梅津知宏、藤本博昭、後藤守孝、赤羽大悟、北原俊彦、吉澤成一郎、大屋敷一馬
    • Organizer
      第74回日本血液学会学術集会
    • Place of Presentation
      京都
    • Related Report
      2012 Annual Research Report
  • [Presentation] 1 分子蛍光分析法によるDNA メチル化度の定量法の開発2011

    • Author(s)
      大屋敷純子、梅津知宏、大屋敷倫代、小林千晶、大屋敷一馬
    • Organizer
      日本人類遺伝学会第56回大会
    • Place of Presentation
      幕張
    • Year and Date
      2011-11-12
    • Related Report
      2012 Final Research Report
  • [Presentation] 非ホジキンリンパ腫の血清 miR-92a レベルは免疫状態と関係し、再発予測因子となる2011

    • Author(s)
      大屋敷倫代、梅津 知宏、吉澤成一郎、大屋敷純子、大屋敷一馬
    • Organizer
      リンパ網内系学会
    • Place of Presentation
      福岡
    • Related Report
      2012 Final Research Report
  • [Presentation] A Novel Global DNA Methylation Detection by Fluorescence Spectroscopy, and Possible Evaluation System of Demethylating Status for Myelodysplastic Patients2011

    • Author(s)
      Ohyashiki K, Umezu T, Ohyashiki JH
    • Organizer
      11th MDS symposium
    • Place of Presentation
      Edinburgh
    • Related Report
      2012 Final Research Report
  • [Presentation] Exosomal miR-92a derived from leukemia cell enhances endothelial cell migration.2011

    • Author(s)
      Umezu T, Ohyashiki JH, Yoshizawa S, Ohyashiki M, Ohyashiki K
    • Organizer
      Keystone Symposia on MicroRNAs and Non-Coding RNAs and Cancer
    • Place of Presentation
      Banff, Canada
    • Related Report
      2011 Research-status Report
  • [Presentation] A Novel Global DNA Methylation Detection by Fluorescence Spectroscopy, and Possible Evaluation System of Demethylating Status for Myelodysplastic Patients.2011

    • Author(s)
      Ohyashiki K, Umezu T, Ohyashiki JH
    • Organizer
      11th MDS symposium
    • Place of Presentation
      Edinburgh
    • Related Report
      2011 Research-status Report
  • [Presentation] DNA demethylation induces up-regulation of telomere repeat-containing RNAs (TERRA) and down-regulation of telomerase activity in human leukemia cells.2011

    • Author(s)
      Ohyashiki JH, Umezu T, Ohyashiki M, Ohtsuki T, Kobayashi C, Ohyashiki K
    • Organizer
      2nd AACR conference on frontiers in basic cancer research
    • Place of Presentation
      San Francisco
    • Related Report
      2011 Research-status Report
  • [Patent(Industrial Property Rights)] 1分子蛍光分析法による DNA メチル化度の決定方法2010

    • Inventor(s)
      大屋敷純子・梅津知宏
    • Industrial Property Rights Holder
      大屋敷一馬
    • Industrial Property Number
      2010-167412
    • Filing Date
      2010-07-26
    • Related Report
      2012 Final Research Report

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Published: 2011-08-05   Modified: 2019-07-29  

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