WO1996035784A2 - Chromatin-regulatorgene - Google Patents
Chromatin-regulatorgene Download PDFInfo
- Publication number
- WO1996035784A2 WO1996035784A2 PCT/EP1996/001818 EP9601818W WO9635784A2 WO 1996035784 A2 WO1996035784 A2 WO 1996035784A2 EP 9601818 W EP9601818 W EP 9601818W WO 9635784 A2 WO9635784 A2 WO 9635784A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- set domain
- cdna
- ezh2
- suv39h
- dna molecule
- Prior art date
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 72
- 108700039010 SET domains Proteins 0.000 claims abstract description 65
- 102000051614 SET domains Human genes 0.000 claims abstract description 58
- 230000003831 deregulation Effects 0.000 claims abstract description 7
- 238000003745 diagnosis Methods 0.000 claims abstract description 6
- 101000882127 Homo sapiens Histone-lysine N-methyltransferase EZH2 Proteins 0.000 claims description 53
- 102100038970 Histone-lysine N-methyltransferase EZH2 Human genes 0.000 claims description 48
- 239000002299 complementary DNA Substances 0.000 claims description 38
- 108010077544 Chromatin Proteins 0.000 claims description 37
- 210000003483 chromatin Anatomy 0.000 claims description 37
- 102100028998 Histone-lysine N-methyltransferase SUV39H1 Human genes 0.000 claims description 26
- 101000696705 Homo sapiens Histone-lysine N-methyltransferase SUV39H1 Proteins 0.000 claims description 25
- 108020004414 DNA Proteins 0.000 claims description 23
- 102000004169 proteins and genes Human genes 0.000 claims description 16
- 230000014509 gene expression Effects 0.000 claims description 12
- 230000036961 partial effect Effects 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 9
- 108700005075 Regulator Genes Proteins 0.000 claims description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 7
- 201000010099 disease Diseases 0.000 claims description 6
- 239000002773 nucleotide Substances 0.000 claims description 6
- 125000003729 nucleotide group Chemical group 0.000 claims description 6
- 108020004511 Recombinant DNA Proteins 0.000 claims description 3
- 210000001671 embryonic stem cell Anatomy 0.000 claims description 2
- 108091006104 gene-regulatory proteins Proteins 0.000 claims description 2
- 102000034356 gene-regulatory proteins Human genes 0.000 claims description 2
- 230000006801 homologous recombination Effects 0.000 claims description 2
- 238000002744 homologous recombination Methods 0.000 claims description 2
- 238000011830 transgenic mouse model Methods 0.000 claims description 2
- 102000053602 DNA Human genes 0.000 claims 10
- 108091028664 Ribonucleotide Proteins 0.000 claims 1
- 108700019146 Transgenes Proteins 0.000 claims 1
- 230000000692 anti-sense effect Effects 0.000 claims 1
- 238000011813 knockout mouse model Methods 0.000 claims 1
- 239000002336 ribonucleotide Substances 0.000 claims 1
- 125000002652 ribonucleotide group Chemical group 0.000 claims 1
- 206010028980 Neoplasm Diseases 0.000 abstract description 8
- 238000002560 therapeutic procedure Methods 0.000 abstract description 4
- 201000011510 cancer Diseases 0.000 abstract description 3
- 102100022103 Histone-lysine N-methyltransferase 2A Human genes 0.000 description 18
- 101001028782 Homo sapiens Histone-lysine N-methyltransferase EZH1 Proteins 0.000 description 18
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 16
- 210000004027 cell Anatomy 0.000 description 16
- 235000018102 proteins Nutrition 0.000 description 14
- 108020004635 Complementary DNA Proteins 0.000 description 13
- 108010041897 SU(VAR)3-9 Proteins 0.000 description 13
- 101001045846 Homo sapiens Histone-lysine N-methyltransferase 2A Proteins 0.000 description 12
- 241000699666 Mus <mouse, genus> Species 0.000 description 12
- 235000001014 amino acid Nutrition 0.000 description 12
- 150000001413 amino acids Chemical class 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 9
- 239000003298 DNA probe Substances 0.000 description 8
- 108050002855 Histone-lysine N-methyltransferase 2A Proteins 0.000 description 8
- 125000003275 alpha amino acid group Chemical group 0.000 description 8
- 230000002074 deregulated effect Effects 0.000 description 8
- 230000035772 mutation Effects 0.000 description 8
- 230000001594 aberrant effect Effects 0.000 description 7
- 230000035755 proliferation Effects 0.000 description 7
- 230000005945 translocation Effects 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 238000012512 characterization method Methods 0.000 description 5
- 230000003993 interaction Effects 0.000 description 5
- 230000002018 overexpression Effects 0.000 description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 4
- 102000017589 Chromo domains Human genes 0.000 description 4
- 108050005811 Chromo domains Proteins 0.000 description 4
- 241000282412 Homo Species 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 238000003780 insertion Methods 0.000 description 4
- 230000037431 insertion Effects 0.000 description 4
- 230000007246 mechanism Effects 0.000 description 4
- 238000003752 polymerase chain reaction Methods 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 4
- 108010035533 Drosophila Proteins Proteins 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 108091028043 Nucleic acid sequence Proteins 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000002759 chromosomal effect Effects 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 238000001415 gene therapy Methods 0.000 description 3
- 102000056255 human EZH2 Human genes 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 108700028369 Alleles Proteins 0.000 description 2
- 108020000948 Antisense Oligonucleotides Proteins 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- 108020003215 DNA Probes Proteins 0.000 description 2
- 108700004507 Drosophila Su3-9 Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 108010034791 Heterochromatin Proteins 0.000 description 2
- 101000836351 Homo sapiens Protein SET Proteins 0.000 description 2
- 101100422860 Homo sapiens SUV39H1 gene Proteins 0.000 description 2
- 108090000144 Human Proteins Proteins 0.000 description 2
- 206010025323 Lymphomas Diseases 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 108060004795 Methyltransferase Proteins 0.000 description 2
- 102000016397 Methyltransferase Human genes 0.000 description 2
- 108020004485 Nonsense Codon Proteins 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 108700026244 Open Reading Frames Proteins 0.000 description 2
- UOZODPSAJZTQNH-UHFFFAOYSA-N Paromomycin II Natural products NC1C(O)C(O)C(CN)OC1OC1C(O)C(OC2C(C(N)CC(N)C2O)OC2C(C(O)C(O)C(CO)O2)N)OC1CO UOZODPSAJZTQNH-UHFFFAOYSA-N 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- 101100477614 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) SIR4 gene Proteins 0.000 description 2
- 238000012300 Sequence Analysis Methods 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 239000012190 activator Substances 0.000 description 2
- 239000000074 antisense oligonucleotide Substances 0.000 description 2
- 238000012230 antisense oligonucleotides Methods 0.000 description 2
- 210000003719 b-lymphocyte Anatomy 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 210000004899 c-terminal region Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- ALEXXDVDDISNDU-JZYPGELDSA-N cortisol 21-acetate Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@@](C(=O)COC(=O)C)(O)[C@@]1(C)C[C@@H]2O ALEXXDVDDISNDU-JZYPGELDSA-N 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 231100000221 frame shift mutation induction Toxicity 0.000 description 2
- 230000037433 frameshift Effects 0.000 description 2
- 230000004545 gene duplication Effects 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 102000049285 human KMT2A Human genes 0.000 description 2
- 102000057013 human SET Human genes 0.000 description 2
- 210000003917 human chromosome Anatomy 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 208000032839 leukemia Diseases 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000003411 telomere Anatomy 0.000 description 2
- 108091035539 telomere Proteins 0.000 description 2
- 102000055501 telomere Human genes 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 230000007704 transition Effects 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 1
- 108020005065 3' Flanking Region Proteins 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 102000008102 Ankyrins Human genes 0.000 description 1
- 108010049777 Ankyrins Proteins 0.000 description 1
- 125000001433 C-terminal amino-acid group Chemical group 0.000 description 1
- 208000037051 Chromosomal Instability Diseases 0.000 description 1
- 108091035707 Consensus sequence Proteins 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- 108700023136 Drosophila Set Proteins 0.000 description 1
- 101000886131 Drosophila melanogaster Transcription factor GAGA Proteins 0.000 description 1
- 108700023507 Drosophila trx Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241001331845 Equus asinus x caballus Species 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 201000001342 Fallopian tube cancer Diseases 0.000 description 1
- 208000013452 Fallopian tube neoplasm Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 108700005087 Homeobox Genes Proteins 0.000 description 1
- 102000009331 Homeodomain Proteins Human genes 0.000 description 1
- 108010048671 Homeodomain Proteins Proteins 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 101000829171 Hypocrea virens (strain Gv29-8 / FGSC 10586) Effector TSP1 Proteins 0.000 description 1
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 1
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 1
- 108700018351 Major Histocompatibility Complex Proteins 0.000 description 1
- 206010064912 Malignant transformation Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 108091008758 NR0A5 Proteins 0.000 description 1
- 238000000636 Northern blotting Methods 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 102000025443 POZ domain binding proteins Human genes 0.000 description 1
- 108091014659 POZ domain binding proteins Proteins 0.000 description 1
- 108700020978 Proto-Oncogene Proteins 0.000 description 1
- 102000052575 Proto-Oncogene Human genes 0.000 description 1
- 108091034057 RNA (poly(A)) Proteins 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 208000036878 aneuploidy Diseases 0.000 description 1
- 231100001075 aneuploidy Toxicity 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000003042 antagnostic effect Effects 0.000 description 1
- 238000000211 autoradiogram Methods 0.000 description 1
- 238000000376 autoradiography Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000001364 causal effect Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 210000002230 centromere Anatomy 0.000 description 1
- 230000008711 chromosomal rearrangement Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000009918 complex formation Effects 0.000 description 1
- 238000010668 complexation reaction Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 230000003412 degenerative effect Effects 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- QTTMOCOWZLSYSV-QWAPEVOJSA-M equilin sodium sulfate Chemical compound [Na+].[O-]S(=O)(=O)OC1=CC=C2[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4C3=CCC2=C1 QTTMOCOWZLSYSV-QWAPEVOJSA-M 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 210000004458 heterochromatin Anatomy 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000001114 immunoprecipitation Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000036212 malign transformation Effects 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000009701 normal cell proliferation Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000037452 priming Effects 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 108020001580 protein domains Proteins 0.000 description 1
- 230000006916 protein interaction Effects 0.000 description 1
- 230000007420 reactivation Effects 0.000 description 1
- 230000010837 receptor-mediated endocytosis Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000001525 retina Anatomy 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 229920006298 saran Polymers 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 description 1
- 206010042863 synovial sarcoma Diseases 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000005748 tumor development Effects 0.000 description 1
- 239000000439 tumor marker Substances 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 239000013603 viral vector Substances 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Classifications
-
- 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
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4702—Regulators; Modulating activity
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A90/00—Technologies having an indirect contribution to adaptation to climate change
- Y02A90/10—Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation
Definitions
- the present invention relates to genes that play a role in the structural and functional regulation of chromatin and their use for therapy and diagnostics.
- telomeres The functional organization of eukaryotic chromosomes in centromeres, telomeres as well as in eu and heterochromatic regions is a crucial mechanism for ensuring the exact replication and distribution of the genetic information in every cell division.
- tumor cells are often characterized by chromosomal rearrangements, translocations and aneuploidy (Solomon et al., 1991; Pardue, 1991).
- this protein domain (Tschiersch et al., 1994), referred to as "SET”, defines a new gene family of mammalian chromatin regulators important in developmental history because of its evolutionary conservation and its presence in antagonistic gene products.
- the characterization of other members of the set of SET domain genes, in addition to HRX / ALL-1, helps to elucidate the mechanisms which are responsible for structural changes in chromatin leading to malignant transformation.
- the object of the present invention was therefore to identify human chromatin regulator genes, to elucidate their function and to use them for diagnosis and therapy.
- the present invention thus relates to DNA molecules containing a sequence coding for a chromatin regulatory protein which has a SET domain or a partial sequence thereof, characterized in that they have the nucleotide sequence shown in FIG. 6 or in FIG. 7.
- the DNA molecules of the invention are also referred to below as "genes of the invention”.
- genes according to the invention have the names EZH2 and SUV39H, they were originally designated as "HEZ-2" and "H3-9".
- the invention relates to the cDNAs derived from these genes, including their degenerate variants; Mutants that code for functional chromatin regulators and variants that are due to gene duplication; an example of this is EZH1, the partial sequence of which is shown in FIG. 8 in comparison with EZH2.
- EZH1 the partial sequence of which is shown in FIG. 8 in comparison with EZH2.
- the procedure was as follows: Starting from the sequence information from the conserved SET domain, a human B-cell-specific cDNA library with a mixed Drosophila ONA probe was used with reduced stringency encoding the SET domains of E (z) and Su (var) 3-9. 40 primary phages were selected from 500,000 plaques.
- the complete cDNA encoding the human homologue of E (z) was named EZH2 and the DNA encoding the human homologue of Su (var) 3-9 was named SUV39H.
- the identity of the amino acids between Drosophila and the human proteins is 61% for EZH2 and 43% for SUV39H, with the C-terminal SET domain being very highly conserved (88% for EZH2 and 53% for SUV39H).
- the sequence comparison showed further clear homology regions, for example a cysteine-rich domain in EZH2 and a chromo box in SÜV39H. (In polycomb it was shown that the chromo box is the domain essential for the interaction between DNA and chromatin; Messmer et al., 1992). In contrast, they are missing 207 amino acids containing the amino-terminal GTP binding motif of the Drosophila protein in the human homologue SUV39H.
- FIGS. 1 and 2 The comparison of the amino acid sequences between the Drosophila and the human genes is shown in FIGS. 1 and 2:
- Fig. 1 shows the comparison of the amino acid sequence between EZH2 and Drosophila Enhancer from zeste (E (z)). Identical amino acids of the conserved carboxy-terminal SET domain (shaded box) and the Cys-rich region (Cys residues are highlighted) are shown. The putative core location signals are underlined.
- FIG. 2 shows the comparison of the amino acid sequence between the human homologue SUV39H and Drosophila Su (var) 3-9. Identical amino acids of the conserved carboxy-terminal SET domain (shaded box) and the chromo domain (darker shaded box) are shown. The putative core location signals are underlined. A schematic overview of the two protein structures is shown at the top of the figure, which shows that 207 amino acids are missing from the N-terminus in the human homologue.
- DNA blot analyzes carried out in the context of the present invention indicate that mammalian homologous genes of Su (var) 3-9 are represented in the mouse and in humans by individual loci, while mammalian homologous genes of E (z) in the mouse and are encoded in humans by two separate loci.
- the second human locus (designated EZH1) was also confirmed by characterization of a small number of cDNA variants, which differ in their 3 'flanking sequences from the majority of the clones isolated from the human cDNA library. The differences between EZH2 and EZH1 in the sequenced region are shown in FIG.
- FIG. 8 shows the nucleotide sequence of the EZH2 cDNA from position 1844 to 2330 in the top line, the 5 1 splice point and the potential stop codon being underlined.
- the Wisconsin GCG network service gap program was used to assign a partial sequence of the cDNA to the EZH1 variant of the _7ZH2 sequence. The premature stop codon in EZHl (position 353) is underlined.
- B52 a partial cDNA sequence with 271 nucleotides called B52, which codes for a mutated variant of the SET domain, was isolated and mapped on the human chromosome 17q21 (Friedman et al., 1994). It was surprisingly found in the context of the present invention that B52 has 97% identity with the BZHL-cDNA variant according to the invention (cf. above); EZH1 may be a gene whose reactivation plays a role in deregulated proliferation.
- a cDNA (2,800 nucleotides; MG-44) had been isolated from the human chromosome Xpll (Geraghty et al., 1993), a region which is responsible for degenerative disorders of the retina and synovial sarcomas predisposed. It was surprisingly found that this cDNA has 98% identity with the SIJV39H-CDNA according to the invention.
- the new genes provided in the context of the present invention thus make it possible to establish a connection between certain cancers and mutations in chromatin regulators; in the case of the G -__- cDNA, since this has several point and frameshift mutations which interrupt the chromo and SET domains, only on the basis of the invention
- G9a is currently the only human gene with a SET domain for which no mutated version is known to date
- KG-1 carries an insertion of 342 amino acids, which cleaves the SET domain into an amino and a carboxy terminal half.
- This JG-1 cDNA is probably an aberrant spliced variant since there are 5 'and 3' consensus splices at both ends of the insertion.
- a total of four of the five currently known human members of the SET protein family are subject to changes, all of which mutate the SET domain (HRX / ALL-1, EZH1 / B52, SUV39H / MG-44 and KG-1).
- the cysteine-rich cluster in B52 is shown as a dotted box; HRX / ALL-1 shows the region of homology to methyltransferases as a hatched box and the A / T hooks as vertical lines. The names of the respective authentic genes are shown on the right in the figure.
- FIG. 4 shows the evolutionary preservation of SET domain proteins: Using the tfasta program of the Wisconsin GCG network service, proteins and open reading frames with homology to the SET domain were identified. A representative selection of yeast to human is shown in the figure. Numbers indicate amino acids.
- the carboxy-terminal SET domain is represented by a black box, Cys-rich regions by a dark-dotted box, the GTP binding motif in Su (var) 3-9 by a light-dotted box and the chromo-domain by Su (var) 3- 9 and H3 -9 through an open box with bright spots.
- a region with homology to methyltransferase (trx and HRX) is shown as a hatched box.
- a / T "hooks" are represented by vertical lines.
- S in C26E6.10 Another Ser-rich region (S in C26E6.10) and a Glu-rich region (E in G9a) or Ankyrin repeats (ANK in G9a) are also highlighted.
- YHR119 GeneBank Accession No. U00059
- C26E6.10 GeneBank Accession No. U13875
- Percentages give the total of Amino acid identities between the human and Drosophila proteins.
- FIG. 5 shows the agreement on amino acids of the SET domain.
- the SET domain of the genes shown in Figure 4 was arranged using the Wisconsin GCG network service pileup program.
- the large amino acid insertion which cleaves the SET domain in half, was removed before the pileup. Amino acid positions that have 8 out of 10 matches are highlighted.
- oligonucleotides coding for the SET domain as such or partial sections thereof can be used as a diagnostic marker in order to diagnose certain types of cancer in which the SET domain is mutated.
- SET domain gene sequences were used to isolate the homologous mouse cDNAs from EZH1 (Ezhl) and SÜV39H (Suv39h).
- RNAse protection analyzes to investigate the JSzhl gene activity during normal mouse development, a rather broad expression profile was found which was similar to that of bmi (Haupt et al. , 1991).
- the analyzes performed with the mouse sequences are expanded with human sequences in order to compare the amounts of RNA between immature precursor cells, tumor cells and differentiated cells in different human cell culture systems.
- RNA concentration can be determined in relevant laboratory manuals (Sambrook, J., Fritsch, EF and Maniatis, T ., 1989, Cold Spring Harbor Laboratory Press), such as Northern blot, Sl nuclease protection analysis or RNAse protection analysis, are used.
- the SET-specific DNA probes can be used for the analysis of single strand conformation polymorphisms (SSCP; Gibbons et al., 1995).
- the present invention thus relates to recombinant DNA molecules containing the DNA coding for EZH2, SUV39H or EZH1 and expression control sequences functionally associated therewith, and the host organisms transformed therewith.
- the recombinant proteins according to the invention can be used to analyze the interaction of SET domain proteins with chromatin or with other members of heterochromatin complexes; On the basis of the knowledge obtained about the mode of action of these complexes, the individual possibilities for the targeted intervention in the mechanisms involved are defined and can be used for therapeutic applications.
- Investigations which serve the further analysis of the function of the SET domain are carried out, for example, by using cDNAs, coding for human EZH2 or SUV39H and provided with an epitope, against the antibody Are available to be expressed in vi tro as well as in tissue cultures. After immunoprecipitation with the respective epitope-specific antibodies, it can be determined whether EZH2 and SUV39H can interact with each other in vitro and whether complexation between EZH2 and / or SUV39H takes place in vivo with further chromatin regulators.
- Heterochromatin proteins are essential for their function. Based on the availability of the SET domain genes according to the invention, it can be determined whether the SET region represents a domain that works due to interactions or whether it contributes to the formation of multimeric heterochromatic complexes. It can also be determined whether the SET domain has an inhibitory function, similar to the amino-terminal BTB domain of various chromatin regulators, including the GAGA factor (Adams et al., 1992). Overall, the analysis of interactions with epitope-labeled EZH2 and _S 3SH proteins allows a further characterization of the function of the SET domain.
- Such variants are obtained, for example, by first defining the functional domains of the SET proteins, for example the sequence segments responsible for the DNA / chromatin or the protein / protein interaction, and then by the domain (s) n) or sections of it shortened DNA sequences are expressed in the affected cell in order to compete for a deregulation of the proliferation caused by the intact functional protein.
- mice are also the subject of the present invention.
- the "gain-of-function” analysis in which alleles of the genes according to the invention are introduced into the mouse, ultimately provides information about the causal involvement of EZH2 and SUV39H in the chromatin-dependent promotion of tumor development.
- the complete cDNA sequences of human EZH2 and SUV39H as well as their mutated versions, such as EZH1 / B52 and MG-44 can be generated using vectors which enable high expression rates, e.g. Plasmids with the human ß-actin promoter driven by both the immunoglobulin heavy chain enhancer (E / x) and Moloney virus enhancers (Mo-LTR).
- DNA sequences according to the invention or sequences derived therefrom are used for gene therapy, depending on whether the disease to be treated is due to deregulation of chromatin as a result of the lack of the functional gene sequence or as a result of overexpression of the corresponding genes.
- introducing the functional gene sequence by inhibiting gene expression, for example with the aid of antisense oligonucleotides, or by introducing a sequence coding for a dominant-negative mutant.
- the respective DNA sequences can be introduced into the cell using standard methods for the transfection of higher eukaryotic cells, to which the gene transfer is carried out by means of viral vectors (retrovirus, adenovirus, adeno-associated virus) or by means of non-viral systems based on the receptor mediated endocytosis count; Overviews of common methods are e.g. by Mitani and Caskey, 1993; Jolly, 1994; Vile and Studssel, 1994; Tepper and Mule, 1994; Zatloukal et al., 1993, WO 93/07283.
- Low-molecular substances which intervene in the transcription machinery can also be considered for inhibiting the expression of the genes according to the invention; after analyzing the The 5 'regulatory region of the genes can be screened for substances which block the interaction of the respective transcription factors with this region in whole or in part, for example using the method described in WO 92/13092.
- An inhibition of the deregulated proliferation can also start at the gene product by using the corresponding antibodies against the EZH2 or SUV39H protein, preferably human or humanized antibodies, therapeutically.
- Antibodies of this type are produced by known methods, such as those e.g. by Malavsi and Albertini, 1992, or by Rhein, 1993.
- Antibodies against EZH2 or SUV39H which can be used therapeutically or diagnostically, are also the subject of the present invention.
- Fig. 1 amino acid sequence comparison between EZH2 and E (z)
- Fig. 2 Amino acid sequence comparison between SÜV39H and Su (var) 3-9
- Fig. 3 Aberrant transcripts from human SET domain-
- Fig. 5 amino acid match in the SET domain
- Fig. 6 EZH2 DNA and amino acid sequence
- Fig. 7 DNA and amino acid sequence of SUV39H
- Fig. 8 partial sequence comparison between the cDNAs of human EZH2 and EZHl example
- Drosophila DNA probes coding for the conserved SET domains of E (z) and Su (var) 3-9, were based on the published Drosophila sequences (Jones and Gelbart, 1993; Tschiersch et al., 1994) prepared by polymerase chain reaction (PCR): 1 ⁇ g of Drosophila melanogaster-ONA (Clontech) was made with the two primers E (z) 1910
- plaque forming units 5 x 10 5 plaque-forming units ( "plaque forming units" pfu) were incubated with 5 ml of the culture of the bacterial host strain E.coli C600 (at an optical density of ODgoo ° - 5 i n 10 M GS0 4 suspended) at 37 ° C incubated for 15 min and then poured onto a large (200 mm x 200 mm) preheated LB dish. After growing overnight at 37 ° C, the phages were absorbed on a nylon membrane (GeneScreen).
- the membrane with the absorbed phage facing upwards, was allowed to float in denaturation solution (1.5 M NaCl, 0.5 M NaOH) for 30 seconds, then immersed in denaturation solution for 60 seconds and finally in 3 M NaCl, 0.5 M Tris for 5 minutes pH 8, neutralized.
- denaturation solution 1.5 M NaCl, 0.5 M NaOH
- 3 M NaCl, 0.5 M Tris for 5 minutes pH 8, neutralized.
- the membrane was then rinsed briefly in 3 ⁇ SSC and the phage DNA was fixed on the nylon filter by UV crosslinking.
- the filter was prehybridized for 30 min at 50 ° C in 30 ml Church buffer (1% BSA, 1 mM EDTA and 0.5 M NaHP0 4 , pH 7.2), then 2 x 10 6 cpm of the radioactively labeled DNA probe mixture (E (z) -SET and Su (var) 3-9-SET) added; the DNA probes were random primed using the RediPrime kit (Amersham). Hybridization was carried out overnight at 50 ° C. After removal of the hybridization solution, the filter was washed in 2xSSC, 1% SDS at room temperature for 10 seconds, and then washed at 50 ° C. for 10 seconds. The filter was wrapped in Saranwrap and subjected to autoradiography using an intensifying screen.
- phage colonies were identified on the original plate using the autoradiogram and remove the corresponding pieces of agar with the larger end of a Pasteur pipette.
- the phage pool was overnight at 4 ° C in 1 ml SM buffer (5.8 g NaCl, 2 g MgS0 4 -H 2 0, 50 ml Tris pH 7.5, 5 ml 2% gelatin on 1 1 H2O) , containing a few drops of CHCI 3 , eluted.
- the phage lysate was replated for a second and third round of screening to obtain individual, well-isolated positive plaques (20 to 100 plaques per plate in the third round).
- phage cDNA inserts were subcloned into the polylinker of pBluescript KS (Stratagene) and sequenced on an automatic sequencer (Applied Biosystems) using the dideoxy method.
- the complete sequence of at least two independent isolates per gene obtained was determined using "primer walking".
- the sequences were analyzed with the GCG software package (University of Wisconsin), the examination for homology was carried out with the "Blast and fasta” or "tfasta” network service.
- the complete sequences of EZH2 and SUV39H are shown in Figs. 6 and 7.
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Toxicology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Saccharide Compounds (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
Claims
Priority Applications (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP53371596A JP4295824B2 (ja) | 1995-05-10 | 1996-05-02 | クロマチン調節遺伝子 |
DK96914165T DK0827537T3 (da) | 1995-05-10 | 1996-05-02 | Chromatin-regulator-gener |
CA2220442A CA2220442C (en) | 1995-05-10 | 1996-05-02 | Chromatin regulator genes |
DE59611481T DE59611481D1 (en) | 1995-05-10 | 1996-05-02 | Chromatin-regulatorgene |
MX9708401A MX9708401A (es) | 1995-05-10 | 1996-05-02 | Genes reguladores de la cromatina. |
EP96914165A EP0827537B1 (de) | 1995-05-10 | 1996-05-02 | Chromatin-regulatorgene |
US10/773,302 US7252968B2 (en) | 1995-05-10 | 2004-02-09 | Chromatin regulator genes |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19516776.7 | 1995-05-10 | ||
DE19516776A DE19516776A1 (de) | 1995-05-10 | 1995-05-10 | Chromatin-Regulatorgene |
Related Child Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US94598897A A-371-Of-International | 1995-05-10 | 1997-11-10 | |
US09/589,892 Continuation-In-Part US6689583B1 (en) | 1995-05-10 | 2000-06-09 | Chromatin regulator genes |
US10/773,302 Continuation-In-Part US7252968B2 (en) | 1995-05-10 | 2004-02-09 | Chromatin regulator genes |
Publications (2)
Publication Number | Publication Date |
---|---|
WO1996035784A2 true WO1996035784A2 (de) | 1996-11-14 |
WO1996035784A3 WO1996035784A3 (de) | 1996-12-27 |
Family
ID=7761318
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP1996/001818 WO1996035784A2 (de) | 1995-05-10 | 1996-05-02 | Chromatin-regulatorgene |
Country Status (11)
Country | Link |
---|---|
US (2) | US6689583B1 (de) |
EP (1) | EP0827537B1 (de) |
JP (1) | JP4295824B2 (de) |
AT (1) | ATE404667T1 (de) |
CA (1) | CA2220442C (de) |
DE (2) | DE19516776A1 (de) |
DK (1) | DK0827537T3 (de) |
ES (1) | ES2313726T3 (de) |
MX (1) | MX9708401A (de) |
PT (1) | PT827537E (de) |
WO (1) | WO1996035784A2 (de) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999053083A1 (en) * | 1998-04-16 | 1999-10-21 | Cold Spring Harbor Laboratory | Seed specific polycomb group gene and methods of use for same |
WO2000053724A2 (en) * | 1999-03-11 | 2000-09-14 | Genesis Research And Development Corporation Ltd | Compositions and methods for the modification of gene transcription |
WO2003055506A1 (fr) * | 2001-12-27 | 2003-07-10 | Takeda Chemical Industries, Ltd. | Medicaments pour la prevention et le traitement du cancer |
EP1577387A1 (de) * | 2002-12-24 | 2005-09-21 | Takeda Pharmaceutical Company Limited | Vorbeugungsmassnahmen/heilmittel für krebs |
US7264964B2 (en) | 2001-06-22 | 2007-09-04 | Ceres, Inc. | Chimeric histone acetyltransferase polypeptides |
US8298816B2 (en) | 2003-12-03 | 2012-10-30 | Chugai Seiyaku Kabushiki Kaisha | Expression systems using mammalian beta-actin promoter |
WO2014153030A2 (en) | 2013-03-14 | 2014-09-25 | Genentech, Inc. | Methods of treating cancer and preventing cancer drug resistance |
Families Citing this family (21)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE19516776A1 (de) | 1995-05-10 | 1996-11-14 | Boehringer Ingelheim Int | Chromatin-Regulatorgene |
US6555329B2 (en) * | 2000-06-09 | 2003-04-29 | Boehringer Ingelheim International Gmbh | Method for identifying compounds altering higher-order chromatin-dependent chromosome stability |
US20020039776A1 (en) | 2000-06-09 | 2002-04-04 | Thomas Jenuwein | Mammalian SUV39H2 proteins and isolated DNA molecules encoding them |
AU2002211624A1 (en) * | 2000-10-10 | 2002-04-22 | Genencor International, Inc. | Information rich libraries |
US20040053848A1 (en) * | 2001-08-23 | 2004-03-18 | Allis C. David | Antibodies specific for methylated lysines in histones |
GB0228900D0 (en) * | 2002-12-11 | 2003-01-15 | Ml Lab Plc | Cancer Immunotherapy |
JP2005170799A (ja) * | 2003-12-08 | 2005-06-30 | Univ Kurume | zesteホモログ2のエンハンサーのHLA−A24結合ペプチド |
ATE449168T1 (de) * | 2004-06-01 | 2009-12-15 | Univ North Carolina | Rekonstituierter histon-methyltransferase-komplex und verfahren zur identifizierung von modulatoren dafür |
EP1987148A4 (de) * | 2006-02-27 | 2009-08-05 | Imgen Co Ltd | Dedifferenzierung von astrozyten zu neuralen stammzellen unter verwendung von bmi-1 |
US8574832B2 (en) | 2010-02-03 | 2013-11-05 | Massachusetts Institute Of Technology | Methods for preparing sequencing libraries |
US20130102477A1 (en) | 2010-06-23 | 2013-04-25 | Ryan D. Morin | Biomarkers for non-hodgkin lymphomas and uses thereof |
US20130195843A1 (en) * | 2010-06-23 | 2013-08-01 | British Columbia Cancer Agency Branch | Biomarkers for Non-Hodgkin Lymphomas and Uses Thereof |
RU2765155C2 (ru) * | 2010-09-10 | 2022-01-26 | Эпизайм, Инк. | Ингибиторы ezh2 человека и способы их применения |
US9175331B2 (en) * | 2010-09-10 | 2015-11-03 | Epizyme, Inc. | Inhibitors of human EZH2, and methods of use thereof |
WO2012050963A2 (en) * | 2010-09-29 | 2012-04-19 | The General Hospital Corporation D/B/A Massachusetts General Hospital | Agents providing controls and standards for immuno-precipitation assays |
TW201733984A (zh) | 2011-04-13 | 2017-10-01 | 雅酶股份有限公司 | 經取代之苯化合物 |
JO3438B1 (ar) | 2011-04-13 | 2019-10-20 | Epizyme Inc | مركبات بنزين مستبدلة بأريل أو أريل غير متجانس |
KR20220123339A (ko) | 2012-04-13 | 2022-09-06 | 에피자임, 인코포레이티드 | 인간 히스톤 메틸트랜스퍼라제 ezh2 억제제의 염 형태 |
MX353929B (es) | 2012-10-15 | 2018-02-02 | Epizyme Inc | Compuestos de benceno sustituidos. |
CN103215352B (zh) * | 2013-03-29 | 2015-03-11 | 中国医学科学院血液病医院(血液学研究所) | 一种检测setd2基因突变的试剂盒 |
LT3057962T (lt) | 2013-10-16 | 2023-12-11 | Epizyme, Inc. | Hidrochlorido druskos forma, skirta ezh2 inhibavimui |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5486623A (en) | 1993-12-08 | 1996-01-23 | Prototek, Inc. | Cysteine protease inhibitors containing heterocyclic leaving groups |
DE19516776A1 (de) | 1995-05-10 | 1996-11-14 | Boehringer Ingelheim Int | Chromatin-Regulatorgene |
JP2001507226A (ja) | 1996-12-20 | 2001-06-05 | ボード オブ リージェンツ,ザ ユニバーシティ オブ テキサス システム | 有糸分裂を調節するためのタンパク質および組成物 |
US6004933A (en) | 1997-04-25 | 1999-12-21 | Cortech Inc. | Cysteine protease inhibitors |
US5972608A (en) | 1997-08-27 | 1999-10-26 | University Of Massachusetts | Assays and reagents for chromatin remodeling enzymes and their modulators |
EP1029547A1 (de) | 1999-02-15 | 2000-08-23 | BOEHRINGER INGELHEIM INTERNATIONAL GmbH | Pharmazeutische wirksame stoffe und Verfahren zu zu ihrer Identifizierung |
US6555329B2 (en) | 2000-06-09 | 2003-04-29 | Boehringer Ingelheim International Gmbh | Method for identifying compounds altering higher-order chromatin-dependent chromosome stability |
US20020039776A1 (en) | 2000-06-09 | 2002-04-04 | Thomas Jenuwein | Mammalian SUV39H2 proteins and isolated DNA molecules encoding them |
EP1227160A1 (de) | 2001-01-19 | 2002-07-31 | BOEHRINGER INGELHEIM INTERNATIONAL GmbH | Verbindungen, die die Trennung von Schwesterchromatiden modulieren, und Verfahren für das Identifizieren dieser Verbindungen |
US20020164620A1 (en) | 2001-01-19 | 2002-11-07 | Boehringer Ingelheim International Gmbh | Method for identifying compounds modulating sister chromatid separation |
CN1678735A (zh) | 2001-05-04 | 2005-10-05 | 佛罗里达大学研究基金会有限公司 | 来源于细菌的丙酮酸脱羧酶(pdc)基因的克隆和测序及其用途 |
-
1995
- 1995-05-10 DE DE19516776A patent/DE19516776A1/de not_active Withdrawn
-
1996
- 1996-05-02 MX MX9708401A patent/MX9708401A/es unknown
- 1996-05-02 DK DK96914165T patent/DK0827537T3/da active
- 1996-05-02 WO PCT/EP1996/001818 patent/WO1996035784A2/de active IP Right Grant
- 1996-05-02 DE DE59611481T patent/DE59611481D1/de not_active Expired - Lifetime
- 1996-05-02 PT PT96914165T patent/PT827537E/pt unknown
- 1996-05-02 CA CA2220442A patent/CA2220442C/en not_active Expired - Lifetime
- 1996-05-02 AT AT96914165T patent/ATE404667T1/de active
- 1996-05-02 ES ES96914165T patent/ES2313726T3/es not_active Expired - Lifetime
- 1996-05-02 EP EP96914165A patent/EP0827537B1/de not_active Expired - Lifetime
- 1996-05-02 JP JP53371596A patent/JP4295824B2/ja not_active Expired - Fee Related
-
2000
- 2000-06-09 US US09/589,892 patent/US6689583B1/en not_active Expired - Lifetime
-
2004
- 2004-02-09 US US10/773,302 patent/US7252968B2/en not_active Expired - Fee Related
Non-Patent Citations (5)
Cited By (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6239327B1 (en) | 1998-04-16 | 2001-05-29 | Cold Spring Harbor Laboratory | Seed specific polycomb group gene and methods of use for same |
WO1999053083A1 (en) * | 1998-04-16 | 1999-10-21 | Cold Spring Harbor Laboratory | Seed specific polycomb group gene and methods of use for same |
US6833446B1 (en) | 1999-03-11 | 2004-12-21 | Agrigenesis Biosciences Limited | Compositions and methods for the modification of gene transcription |
WO2000053724A3 (en) * | 1999-03-11 | 2000-12-28 | Genesis Res & Dev Corp Ltd | Compositions and methods for the modification of gene transcription |
WO2000053724A2 (en) * | 1999-03-11 | 2000-09-14 | Genesis Research And Development Corporation Ltd | Compositions and methods for the modification of gene transcription |
US7264964B2 (en) | 2001-06-22 | 2007-09-04 | Ceres, Inc. | Chimeric histone acetyltransferase polypeptides |
WO2003055506A1 (fr) * | 2001-12-27 | 2003-07-10 | Takeda Chemical Industries, Ltd. | Medicaments pour la prevention et le traitement du cancer |
EP1459754A1 (de) * | 2001-12-27 | 2004-09-22 | Takeda Chemical Industries, Ltd. | Mittel zur verhütung/behandlung von krebs |
EP1459754A4 (de) * | 2001-12-27 | 2005-08-31 | Takeda Pharmaceutical | Mittel zur verhütung/behandlung von krebs |
EP1577387A1 (de) * | 2002-12-24 | 2005-09-21 | Takeda Pharmaceutical Company Limited | Vorbeugungsmassnahmen/heilmittel für krebs |
EP1577387A4 (de) * | 2002-12-24 | 2006-02-15 | Takeda Pharmaceutical | Vorbeugungsmassnahmen/heilmittel für krebs |
US8298816B2 (en) | 2003-12-03 | 2012-10-30 | Chugai Seiyaku Kabushiki Kaisha | Expression systems using mammalian beta-actin promoter |
WO2014153030A2 (en) | 2013-03-14 | 2014-09-25 | Genentech, Inc. | Methods of treating cancer and preventing cancer drug resistance |
Also Published As
Publication number | Publication date |
---|---|
US20050089880A1 (en) | 2005-04-28 |
CA2220442A1 (en) | 1996-11-14 |
CA2220442C (en) | 2011-07-12 |
PT827537E (pt) | 2008-10-09 |
US6689583B1 (en) | 2004-02-10 |
JP2001505402A (ja) | 2001-04-24 |
DK0827537T3 (da) | 2008-12-01 |
ATE404667T1 (de) | 2008-08-15 |
EP0827537A2 (de) | 1998-03-11 |
DE19516776A1 (de) | 1996-11-14 |
MX9708401A (es) | 1998-02-28 |
JP4295824B2 (ja) | 2009-07-15 |
ES2313726T3 (es) | 2009-03-01 |
EP0827537B1 (de) | 2008-08-13 |
US7252968B2 (en) | 2007-08-07 |
WO1996035784A3 (de) | 1996-12-27 |
DE59611481D1 (en) | 2008-09-25 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP0827537B1 (de) | Chromatin-regulatorgene | |
DE60019273T2 (de) | Verwendung von casb616 polypeptiden und polynukleotiden zur krebsbehandlung | |
DE69430808T2 (de) | Nf-at-polypeptide und-polynukleotide | |
MXPA97008401A (en) | Regulatory genes of the cromat | |
DE69434614T2 (de) | Cdn apoptosis modulierende proteine, sie kodierende dns und verfahren zu ihrer verwendung | |
DE69534351T2 (de) | Motorneuronlebenserhaltungsgen: ein Gen für spinale Muskelatrophie | |
DE69534733T2 (de) | NEUARTIGE AUF p53 ANSPRECHENDE GENE | |
US5773268A (en) | Chromosome 21 gene marker, compositions and methods using same | |
DE69635349T2 (de) | Nukleotidsequenzen, proteine, medikamente und diagnoseagentien zur anwendung in krebsbehandlung | |
DE69332798T2 (de) | Ikaros: ein regulatorisches gen bei der t-zell differenzierung | |
Yamakawa et al. | Isolation and characterization of a candidate gene for progressive myoclonus epilepsy on 21q22. 3 | |
DE60128830T2 (de) | Regulierung der expression von eer-7, ein mitglied der lysyl oxidase genfamilie, durch oestrogenrezeptoren | |
DE69636870T2 (de) | KVLQT1 - EIN GEN DES LONG-QT-SYNDROMS, WELCHES FÜR KVLQT1 KODIERT UND WELCHES SICH MIT Mink ZUSAMMENSETZT, UM Iks-KALIUMKANÄLE DES HERZENS ZU BILDEN | |
DE69934239T2 (de) | Ly6h-gen | |
DE69735604T2 (de) | Hypophysentumor verursachende gene und verwandte produkte | |
DE69718052T2 (de) | Menschliches wachstumsgen und minderwuchse gen bereich | |
DE69534753T2 (de) | Nierenpolycystose-gen | |
DE69838565T2 (de) | Nukleinsäuren von CIITA Genen | |
WO1998045428A1 (de) | Fanconi-gen ii | |
DE60024862T2 (de) | "insulinabhängiges sequenz dna bindendes protein-1" (irsdbp-1), dafür kodierendes gen und ihre verwendungen | |
WO2001032693A2 (de) | Mit trp-proteinen verwandtes protein mtr1 und dieses codierende dna-sequenz | |
EP1220921A1 (de) | Nukleinsäuresequenzen von hyperplasien und tumoren der schilddrüse | |
DE19729211C2 (de) | Humanes Semaphorin L (H-Sema-L) und korrespondierende Semaphorine in anderen Spezies | |
DE10001377A1 (de) | bHLH-Transkriptionsfaktor ASCL3 und diesen enthaltendes Arzneimittel | |
AT410945B (de) | Verfahren zur diagnose von multipler sklerose (ms) |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AK | Designated states |
Kind code of ref document: A2 Designated state(s): CA JP MX US |
|
AL | Designated countries for regional patents |
Kind code of ref document: A2 Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE |
|
AK | Designated states |
Kind code of ref document: A3 Designated state(s): CA JP MX US |
|
AL | Designated countries for regional patents |
Kind code of ref document: A3 Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE |
|
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
WWE | Wipo information: entry into national phase |
Ref document number: PA/a/1997/008401 Country of ref document: MX |
|
ENP | Entry into the national phase |
Ref document number: 2220442 Country of ref document: CA Ref country code: CA Ref document number: 2220442 Kind code of ref document: A Format of ref document f/p: F |
|
ENP | Entry into the national phase |
Ref country code: US Ref document number: 1997 945988 Date of ref document: 19971110 Kind code of ref document: A Format of ref document f/p: F Ref country code: JP Ref document number: 1996 533715 Kind code of ref document: A Format of ref document f/p: F |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1996914165 Country of ref document: EP |
|
WWP | Wipo information: published in national office |
Ref document number: 1996914165 Country of ref document: EP |
|
WWG | Wipo information: grant in national office |
Ref document number: 1996914165 Country of ref document: EP |