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WO2013111515A1 - Procédé d'induction/activation d'une cellule souche/progénitrice cardiaque à l'aide d'un facteur spécifique - Google Patents

Procédé d'induction/activation d'une cellule souche/progénitrice cardiaque à l'aide d'un facteur spécifique Download PDF

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Publication number
WO2013111515A1
WO2013111515A1 PCT/JP2012/084259 JP2012084259W WO2013111515A1 WO 2013111515 A1 WO2013111515 A1 WO 2013111515A1 JP 2012084259 W JP2012084259 W JP 2012084259W WO 2013111515 A1 WO2013111515 A1 WO 2013111515A1
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Prior art keywords
seq
gene
protein
nucleotide sequence
cells
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Ceased
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PCT/JP2012/084259
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English (en)
Japanese (ja)
Inventor
竹内　純
唯加 森田
由布子 塚原
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University of Tokyo NUC
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University of Tokyo NUC
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Publication of WO2013111515A1 publication Critical patent/WO2013111515A1/fr
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0657—Cardiomyocytes; Heart cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
    • C12N2506/02—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from embryonic cells

Definitions

  • FIG. 15 is a diagram showing that typical cardiac muscle markers such as PDGFRa and cKit are also expressed in cells in which gene C is expressed.
  • FIG. 16 is a diagram showing that differentiation into myocardium proceeds in inverse proportion to the relative expression level of gene C expressed in cells.
  • FIG. 17 shows that the area of cardiac progenitor cells in the developing heart extends over a wider area than previously known.
  • FIG. 18 shows that gene C is expressed in cardiac endothelial cells.
  • FIG. 19 shows gene C, gene I and gene K expressed exogenously in the embryo.
  • Proteins belonging to Family Z include protein J (SEQ ID NO: 18 amino acid sequence, SEQ ID NO: 20 amino acid sequence, or similar amino acid sequence) or protein K (SEQ ID NO: 22 amino acid sequence). Sequence, SEQ ID NO: 24 amino acid sequence, or an amino acid sequence similar thereto), among them, protein K (SEQ ID NO: 24), which is an amino acid sequence derived from a human, and the like. It is preferable to use gene K (SEQ ID NO: 23).
  • the stringent conditions used in the present specification can be easily determined by those skilled in the art having general techniques based on, for example, the length of DNA.
  • the basic conditions are Sambrook et al. , Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory Press, (2001).
  • moderately stringent conditions for nitrocellulose filters are 5 ⁇ SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0) pre-wash solution, about 40 ° C. to 60 ° C., preferably about Other similar hybridization solutions, such as Stark's solution, in 1 ⁇ SSC to 6 ⁇ SSC, preferably 2 ⁇ SSC (or about 50% formamide at about 42 ° C. at 50 ° C.
  • the gene H and gene I belonging to the family Y of the present invention are strongly expressed in the early fetal period and disappear after the myocardial differentiation period.
  • a factor belonging to family X protein C or protein D, or gene C or gene D encoding them
  • a factor belonging to family Y protein H or protein I, or gene H or genes encoding them
  • production of marker molecules generally known as cardiomyocyte markers
  • a strong expression group that expresses gene C was collected from the cell group that expresses gene C, and the following examination was performed.
  • a transgenic mouse gene C-GFP knock-in mouse
  • EGFP fluorescent tag
  • Cell groups ie, gene C + cells
  • the cells are expanded with embryonic cardiac progenitor cell-specific cell surface antigens c-kit and PDGFRa (The upper right figure of FIG. 15).
  • SEQ ID NO: 5 nucleotide sequence encoding mouse-derived gene D
  • SEQ ID NO: 22 amino acid sequence of mouse-derived gene K

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Wood Science & Technology (AREA)
  • Cardiology (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Toxicology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Public Health (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Rheumatology (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Engineering & Computer Science (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
PCT/JP2012/084259 2012-01-23 2012-12-21 Procédé d'induction/activation d'une cellule souche/progénitrice cardiaque à l'aide d'un facteur spécifique Ceased WO2013111515A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2012011458A JP2015078126A (ja) 2012-01-23 2012-01-23 高効率心臓細胞分化能を持った新規心臓前駆（幹）細胞制御因子
JP2012-011458 2012-01-23

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WO2013111515A1 true WO2013111515A1 (fr) 2013-08-01

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JP (1) JP2015078126A (fr)
WO (1) WO2013111515A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2017060422A (ja) * 2015-09-24 2017-03-30 国立大学法人 東京大学 成熟した心筋細胞を分化誘導させる方法

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010042800A1 (fr) * 2008-10-10 2010-04-15 Nevada Cancer Institute Procédés de reprogrammation de cellules somatiques et procédés d'utilisation de telles cellules

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010042800A1 (fr) * 2008-10-10 2010-04-15 Nevada Cancer Institute Procédés de reprogrammation de cellules somatiques et procédés d'utilisation de telles cellules

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
BOEHM J. ET AL.: "Synergistic cooperation of Sall4 and Cyclin D1 in transcriptional repression", BIOCHEM. BIOPHYS. RES. COMMUN., vol. 356, no. 3, 11 May 2007 (2007-05-11), pages 773 - 779, XP022006055 *
GUO C. ET AL.: "A Tbx1-Six1/Eyal-Fgf8 genetic pathway controls mammalian cardiovascular and craniofacial morphogenesis", J. CLIN. INVEST., vol. 121, no. 4, April 2011 (2011-04-01), pages 1585 - 1595, XP055081117 *
KUME T. ET AL.: "The murine winged helix transcription factors, Foxc1 and Foxc2, are both required for cardiovascular development and somitogenesis", GENES DEV., vol. 15, no. 18, 15 September 2001 (2001-09-15), pages 2470 - 2482, XP002962537 *
LAUGWITZ K.L. ET AL.: "Islet1 cardiovascular progenitors: a single source for heart lineages?", DEVELOPMENT, vol. 135, no. 2, January 2008 (2008-01-01), pages 193 - 205, XP002491238 *
SASAKI-YUMOTO M. ET AL.: "The murine homolog of SALL4, a causative gene in Okihiro syndrome, is essential for embryonic stem cell proliferation, and cooperates with Sall1 in anorectal, heart, brain and kidney development", DEVELOPMENT, vol. 133, no. 15, August 2006 (2006-08-01), pages 3005 - 3013, XP055081114 *
SCHOENBERGER J. ET AL.: "Mutation in the transcriptional coactivator EYA4 causes dilated cardiomyopathy and sensorineural hearing loss", NAT. GENET., vol. 37, no. 4, April 2005 (2005-04-01), pages 418 - 422, XP055081121 *
SEO S. ET AL.: "Forkhead transcription factors, Foxc1 and Foxc2, are required for the morphogenesis of the cardiac outflow tract", DEV. BIOL., vol. 296, no. 2, 15 August 2006 (2006-08-15), pages 421 - 436, XP005597097 *
YUIKA MORITA ET AL.: "Aratana Shinzo Zenkusaibo Seigyo Inshi to Kaisosei no Rikai", KEKKAN, vol. 35, no. 1, 31 January 2012 (2012-01-31), pages 37 *
YUIKA MORITA ET AL.: "Sall wa Shinzo Zenkusaibo Hissu Inshi to shite Zen Shinzo Saibo Keifu o Seigyo suru", REGENERATIVE MEDICINE, vol. 11, 16 May 2012 (2012-05-16) *

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