JPWO2020149395A1 - 栄養障害型表皮水疱症治療薬 - Google Patents
栄養障害型表皮水疱症治療薬 Download PDFInfo
- Publication number
- JPWO2020149395A1 JPWO2020149395A1 JP2020566497A JP2020566497A JPWO2020149395A1 JP WO2020149395 A1 JPWO2020149395 A1 JP WO2020149395A1 JP 2020566497 A JP2020566497 A JP 2020566497A JP 2020566497 A JP2020566497 A JP 2020566497A JP WO2020149395 A1 JPWO2020149395 A1 JP WO2020149395A1
- Authority
- JP
- Japan
- Prior art keywords
- cells
- cell
- acid sequence
- composition according
- mesenchymal stem
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 206010014989 Epidermolysis bullosa Diseases 0.000 title claims abstract description 36
- 239000003814 drug Substances 0.000 title description 5
- 229940079593 drug Drugs 0.000 title 1
- 210000004027 cell Anatomy 0.000 claims abstract description 263
- 239000000203 mixture Substances 0.000 claims abstract description 90
- 108010017377 Collagen Type VII Proteins 0.000 claims abstract description 62
- 102000004510 Collagen Type VII Human genes 0.000 claims abstract description 61
- 208000010975 Dystrophic epidermolysis bullosa Diseases 0.000 claims abstract description 16
- 208000004298 epidermolysis bullosa dystrophica Diseases 0.000 claims abstract description 16
- 210000002901 mesenchymal stem cell Anatomy 0.000 claims description 67
- 101150056204 COL7A1 gene Proteins 0.000 claims description 59
- 101100496573 Homo sapiens COL7A1 gene Proteins 0.000 claims description 59
- 150000007523 nucleic acids Chemical group 0.000 claims description 40
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 34
- 108020005004 Guide RNA Proteins 0.000 claims description 19
- 210000001185 bone marrow Anatomy 0.000 claims description 14
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 13
- 230000001771 impaired effect Effects 0.000 claims description 11
- 230000000295 complement effect Effects 0.000 claims description 7
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 208000015380 nutritional deficiency disease Diseases 0.000 abstract description 11
- 210000000130 stem cell Anatomy 0.000 abstract description 3
- 239000013598 vector Substances 0.000 description 58
- 238000000034 method Methods 0.000 description 40
- 108091033409 CRISPR Proteins 0.000 description 21
- 150000001413 amino acids Chemical class 0.000 description 21
- 238000010362 genome editing Methods 0.000 description 20
- 210000003491 skin Anatomy 0.000 description 16
- 239000013603 viral vector Substances 0.000 description 15
- 108010042407 Endonucleases Proteins 0.000 description 14
- 241000700605 Viruses Species 0.000 description 13
- 230000035772 mutation Effects 0.000 description 13
- 210000004207 dermis Anatomy 0.000 description 12
- 238000002347 injection Methods 0.000 description 12
- 239000007924 injection Substances 0.000 description 12
- 108090000623 proteins and genes Proteins 0.000 description 12
- 238000004806 packaging method and process Methods 0.000 description 11
- 239000013612 plasmid Substances 0.000 description 11
- 102000008186 Collagen Human genes 0.000 description 10
- 108010035532 Collagen Proteins 0.000 description 10
- 102100031780 Endonuclease Human genes 0.000 description 10
- 210000002469 basement membrane Anatomy 0.000 description 10
- 229920001436 collagen Polymers 0.000 description 10
- 238000012239 gene modification Methods 0.000 description 10
- 238000010354 CRISPR gene editing Methods 0.000 description 9
- 210000002615 epidermis Anatomy 0.000 description 9
- 108010061833 Integrases Proteins 0.000 description 8
- 230000005017 genetic modification Effects 0.000 description 8
- 235000013617 genetically modified food Nutrition 0.000 description 8
- 102100034343 Integrase Human genes 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 6
- 208000002720 Malnutrition Diseases 0.000 description 6
- 238000004873 anchoring Methods 0.000 description 6
- 238000003776 cleavage reaction Methods 0.000 description 6
- 210000002510 keratinocyte Anatomy 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 108020004707 nucleic acids Proteins 0.000 description 6
- 102000039446 nucleic acids Human genes 0.000 description 6
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 6
- 230000001177 retroviral effect Effects 0.000 description 6
- 230000007017 scission Effects 0.000 description 6
- 210000001519 tissue Anatomy 0.000 description 6
- 241000702421 Dependoparvovirus Species 0.000 description 5
- 102100035102 E3 ubiquitin-protein ligase MYCBP2 Human genes 0.000 description 5
- 101000909498 Homo sapiens Collagen alpha-1(VII) chain Proteins 0.000 description 5
- 108091027544 Subgenomic mRNA Proteins 0.000 description 5
- 239000002299 complementary DNA Substances 0.000 description 5
- 230000002950 deficient Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 210000002950 fibroblast Anatomy 0.000 description 5
- 238000012744 immunostaining Methods 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 102000004533 Endonucleases Human genes 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 4
- 230000001071 malnutrition Effects 0.000 description 4
- 235000000824 malnutrition Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 238000001890 transfection Methods 0.000 description 4
- 238000011144 upstream manufacturing Methods 0.000 description 4
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 3
- 108700028369 Alleles Proteins 0.000 description 3
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 3
- 102100024335 Collagen alpha-1(VII) chain Human genes 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 241000725303 Human immunodeficiency virus Species 0.000 description 3
- 241000713666 Lentivirus Species 0.000 description 3
- 108091028113 Trans-activating crRNA Proteins 0.000 description 3
- 125000000539 amino acid group Chemical group 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 3
- 238000004520 electroporation Methods 0.000 description 3
- 239000000835 fiber Substances 0.000 description 3
- 239000002245 particle Substances 0.000 description 3
- 239000013600 plasmid vector Substances 0.000 description 3
- 229950010131 puromycin Drugs 0.000 description 3
- 102220032988 rs281865408 Human genes 0.000 description 3
- 229940124597 therapeutic agent Drugs 0.000 description 3
- 241001430294 unidentified retrovirus Species 0.000 description 3
- 108700001624 vesicular stomatitis virus G Proteins 0.000 description 3
- 241000486679 Antitype Species 0.000 description 2
- 241000713756 Caprine arthritis encephalitis virus Species 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 229920002307 Dextran Polymers 0.000 description 2
- 206010059866 Drug resistance Diseases 0.000 description 2
- 241000713730 Equine infectious anemia virus Species 0.000 description 2
- 241000713800 Feline immunodeficiency virus Species 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 208000028782 Hereditary disease Diseases 0.000 description 2
- 101000800116 Homo sapiens Thy-1 membrane glycoprotein Proteins 0.000 description 2
- 208000026350 Inborn Genetic disease Diseases 0.000 description 2
- 208000024556 Mendelian disease Diseases 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 241000713311 Simian immunodeficiency virus Species 0.000 description 2
- 241000193996 Streptococcus pyogenes Species 0.000 description 2
- 102100033523 Thy-1 membrane glycoprotein Human genes 0.000 description 2
- 108020000999 Viral RNA Proteins 0.000 description 2
- 241000713325 Visna/maedi virus Species 0.000 description 2
- 101150063416 add gene Proteins 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 239000000853 adhesive Substances 0.000 description 2
- 230000001070 adhesive effect Effects 0.000 description 2
- 210000001789 adipocyte Anatomy 0.000 description 2
- -1 as well as skin Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 210000001612 chondrocyte Anatomy 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000003623 enhancer Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000001415 gene therapy Methods 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 210000000963 osteoblast Anatomy 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 239000011886 peripheral blood Substances 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 239000004033 plastic Substances 0.000 description 2
- 229920003023 plastic Polymers 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 241000701161 unidentified adenovirus Species 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- RNAMYOYQYRYFQY-UHFFFAOYSA-N 2-(4,4-difluoropiperidin-1-yl)-6-methoxy-n-(1-propan-2-ylpiperidin-4-yl)-7-(3-pyrrolidin-1-ylpropoxy)quinazolin-4-amine Chemical compound N1=C(N2CCC(F)(F)CC2)N=C2C=C(OCCCN3CCCC3)C(OC)=CC2=C1NC1CCN(C(C)C)CC1 RNAMYOYQYRYFQY-UHFFFAOYSA-N 0.000 description 1
- 102100022464 5'-nucleotidase Human genes 0.000 description 1
- 241000093740 Acidaminococcus sp. Species 0.000 description 1
- 102100032912 CD44 antigen Human genes 0.000 description 1
- 108091079001 CRISPR RNA Proteins 0.000 description 1
- 101100348617 Candida albicans (strain SC5314 / ATCC MYA-2876) NIK1 gene Proteins 0.000 description 1
- 108700004991 Cas12a Proteins 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 238000000116 DAPI staining Methods 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 208000031886 HIV Infections Diseases 0.000 description 1
- 101000678236 Homo sapiens 5'-nucleotidase Proteins 0.000 description 1
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 1
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 description 1
- 101000935043 Homo sapiens Integrin beta-1 Proteins 0.000 description 1
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 1
- 101000687905 Homo sapiens Transcription factor SOX-2 Proteins 0.000 description 1
- 101000835093 Homo sapiens Transferrin receptor protein 1 Proteins 0.000 description 1
- 241000713772 Human immunodeficiency virus 1 Species 0.000 description 1
- 241000713340 Human immunodeficiency virus 2 Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102100022338 Integrin alpha-M Human genes 0.000 description 1
- 102100025304 Integrin beta-1 Human genes 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 241000689670 Lachnospiraceae bacterium ND2006 Species 0.000 description 1
- 241000588650 Neisseria meningitidis Species 0.000 description 1
- 108010077850 Nuclear Localization Signals Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 102100035423 POU domain, class 5, transcription factor 1 Human genes 0.000 description 1
- 101710126211 POU domain, class 5, transcription factor 1 Proteins 0.000 description 1
- 102100024616 Platelet endothelial cell adhesion molecule Human genes 0.000 description 1
- 102000001393 Platelet-Derived Growth Factor alpha Receptor Human genes 0.000 description 1
- 108010068588 Platelet-Derived Growth Factor alpha Receptor Proteins 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 1
- 102000018120 Recombinases Human genes 0.000 description 1
- 108010091086 Recombinases Proteins 0.000 description 1
- 101100007329 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) COS1 gene Proteins 0.000 description 1
- 101100221606 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) COS7 gene Proteins 0.000 description 1
- 108091058545 Secretory proteins Proteins 0.000 description 1
- 102000040739 Secretory proteins Human genes 0.000 description 1
- 206010040943 Skin Ulcer Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 238000002105 Southern blotting Methods 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 238000010459 TALEN Methods 0.000 description 1
- 108010043645 Transcription Activator-Like Effector Nucleases Proteins 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 102100024270 Transcription factor SOX-2 Human genes 0.000 description 1
- 102100026144 Transferrin receptor protein 1 Human genes 0.000 description 1
- 241000589892 Treponema denticola Species 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 102000016549 Vascular Endothelial Growth Factor Receptor-2 Human genes 0.000 description 1
- 108010053099 Vascular Endothelial Growth Factor Receptor-2 Proteins 0.000 description 1
- 208000010094 Visna Diseases 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000000735 allogeneic effect Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 210000000270 basal cell Anatomy 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 210000003969 blast cell Anatomy 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 208000019748 bullous skin disease Diseases 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000002577 cryoprotective agent Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 230000009429 distress Effects 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000005782 double-strand break Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 230000001605 fetal effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 201000005787 hematologic cancer Diseases 0.000 description 1
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 210000003692 ilium Anatomy 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- 208000008106 junctional epidermolysis bullosa Diseases 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 238000001638 lipofection Methods 0.000 description 1
- 238000002826 magnetic-activated cell sorting Methods 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 238000004886 process control Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 238000007390 skin biopsy Methods 0.000 description 1
- 210000004927 skin cell Anatomy 0.000 description 1
- 210000001626 skin fibroblast Anatomy 0.000 description 1
- 231100000019 skin ulcer Toxicity 0.000 description 1
- 238000010583 slow cooling Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 230000037436 splice-site mutation Effects 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 210000001562 sternum Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 210000002303 tibia Anatomy 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 210000000605 viral structure Anatomy 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/02—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
-
- 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
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/90—Stable introduction of foreign DNA into chromosome
- C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
- C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian 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
- 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
- C12N5/0662—Stem cells
- C12N5/0663—Bone marrow mesenchymal stem cells (BM-MSC)
-
- 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
- A01K2217/07—Animals genetically altered by homologous recombination
- A01K2217/075—Animals genetically altered by homologous recombination inducing loss of function, i.e. knock out
-
- 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
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- 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
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/0306—Animal model for genetic diseases
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- 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
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPRs]
-
- 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
- C12N2510/00—Genetically modified cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Cell Biology (AREA)
- Developmental Biology & Embryology (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- Epidemiology (AREA)
- General Engineering & Computer Science (AREA)
- Dermatology (AREA)
- General Chemical & Material Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Hematology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Gastroenterology & Hepatology (AREA)
- Physics & Mathematics (AREA)
- Plant Pathology (AREA)
- Virology (AREA)
- Mycology (AREA)
- Toxicology (AREA)
- Rheumatology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
本開示は、栄養障害型表皮水疱症の治療薬に関する。
ヒトCOL7A1遺伝子のcDNA配列(8835 bp)(配列番号1)
ATGACGCTGCGGCTTCTGGTGGCCGCGCTCTGCGCCGGGATCCTGGCAGAGGCGCCCCGAGTGCGAGCCCAGCACAGGGAGAGAGTGACCTGCACGCGCCTTTACGCCGCTGACATTGTGTTCTTACTGGATGGCTCCTCATCCATTGGCCGCAGCAATTTCCGCGAGGTCCGCAGCTTTCTCGAAGGGCTGGTGCTGCCTTTCTCTGGAGCAGCCAGTGCACAGGGTGTGCGCTTTGCCACAGTGCAGTACAGCGATGATCCACGGACAGAGTTCGGCCTGGATGCACTTGGCTCTGGGGGTGATGTGATCCGCGCCATCCGTGAGCTTAGCTACAAGGGGGGCAACACTCGCACAGGGGCTGCAATTCTCCATGTGGCTGACCATGTCTTCCTGCCCCAGCTGGCCCGACCTGGTGTCCCCAAGGTCTGCATCCTGATCACAGACGGGAAGTCCCAGGACCTGGTGGACACAGCTGCCCAAAGGCTGAAGGGGCAGGGGGTCAAGCTATTTGCTGTGGGGATCAAGAATGCTGACCCTGAGGAGCTGAAGCGAGTTGCCTCACAGCCCACCAGTGACTTCTTCTTCTTCGTCAATGACTTCAGCATCTTGAGGACACTACTGCCCCTCGTTTCCCGGAGAGTGTGCACGACTGCTGGTGGCGTGCCTGTGACCCGACCTCCGGATGACTCGACCTCTGCTCCACGAGACCTGGTGCTGTCTGAGCCAAGCAGCCAATCCTTGAGAGTACAGTGGACAGCGGCCAGTGGCCCTGTGACTGGCTACAAGGTCCAGTACACTCCTCTGACGGGGCTGGGACAGCCACTGCCGAGTGAGCGGCAGGAGGTGAACGTCCCAGCTGGTGAGACCAGTGTGCGGCTGCGGGGTCTCCGGCCACTGACCGAGTACCAAGTGACTGTGATTGCCCTCTACGCCAACAGCATCGGGGAGGCTGTGAGCGGGACAGCTCGGACCACTGCCCTAGAAGGGCCGGAACTGACCATCCAGAATACCACAGCCCACAGCCTCCTGGTGGCCTGGCGGAGTGTGCCAGGTGCCACTGGCTACCGTGTGACATGGCGGGTCCTCAGTGGTGGGCCCACACAGCAGCAGGAGCTGGGCCCTGGGCAGGGTTCAGTGTTGCTGCGTGACTTGGAGCCTGGCACGGACTATGAGGTGACCGTGAGCACCCTATTTGGCCGCAGTGTGGGGCCCGCCACTTCCCTGATGGCTCGCACTGACGCTTCTGTTGAGCAGACCCTGCGCCCGGTCATCCTGGGCCCCACATCCATCCTCCTTTCCTGGAACTTGGTGCCTGAGGCCCGTGGCTACCGGTTGGAATGGCGGCGTGAGACTGGCTTGGAGCCACCGCAGAAGGTGGTACTGCCCTCTGATGTGACCCGCTACCAGTTGGATGGGCTGCAGCCGGGCACTGAGTACCGCCTCACACTCTACACTCTGCTGGAGGGCCACGAGGTGGCCACCCCTGCAACCGTGGTTCCCACTGGACCAGAGCTGCCTGTGAGCCCTGTAACAGACCTGCAAGCCACCGAGCTGCCCGGGCAGCGGGTGCGAGTGTCCTGGAGCCCAGTCCCTGGTGCCACCCAGTACCGCATCATTGTGCGCAGCACCCAGGGGGTTGAGCGGACCCTGGTGCTTCCTGGGAGTCAGACAGCATTCGACTTGGATGACGTTCAGGCTGGGCTTAGCTACACTGTGCGGGTGTCTGCTCGAGTGGGTCCCCGTGAGGGCAGTGCCAGTGTCCTCACTGTCCGCCGGGAGCCGGAAACTCCACTTGCTGTTCCAGGGCTGCGGGTTGTGGTGTCAGATGCAACGCGAGTGAGGGTGGCCTGGGGACCCGTCCCTGGAGCCAGTGGATTTCGGATTAGCTGGAGCACAGGCAGTGGTCCGGAGTCCAGCCAGACACTGCCCCCAGACTCTACTGCCACAGACATCACAGGGCTGCAGCCTGGAACCACCTACCAGGTGGCTGTGTCGGTACTGCGAGGCAGAGAGGAGGGCCCTGCTGCAGTCATCGTGGCTCGAACGGACCCACTGGGCCCAGTGAGGACGGTCCATGTGACTCAGGCCAGCAGCTCATCTGTCACCATTACCTGGACCAGGGTTCCTGGCGCCACAGGATACAGGGTTTCCTGGCACTCAGCCCACGGCCCAGAGAAATCCCAGTTGGTTTCTGGGGAGGCCACGGTGGCTGAGCTGGATGGACTGGAGCCAGATACTGAGTATACGGTGCATGTGAGGGCCCATGTGGCTGGCGTGGATGGGCCCCCTGCCTCTGTGGTTGTGAGGACTGCCCCTGAGCCTGTGGGTCGTGTGTCGAGGCTGCAGATCCTCAATGCTTCCAGCGACGTTCTACGGATCACCTGGGTAGGGGTCACTGGAGCCACAGCTTACAGACTGGCCTGGGGCCGGAGTGAAGGCGGCCCCATGAGGCACCAGATACTCCCAGGAAACACAGACTCTGCAGAGATCCGGGGTCTCGAAGGTGGAGTCAGCTACTCAGTGCGAGTGACTGCACTTGTCGGGGACCGCGAGGGCACACCTGTCTCCATTGTTGTCACTACGCCGCCTGAGGCTCCGCCAGCCCTGGGGACGCTTCACGTGGTGCAGCGCGGGGAGCACTCGCTGAGGCTGCGCTGGGAGCCGGTGCCCAGAGCGCAGGGCTTCCTTCTGCACTGGCAACCTGAGGGTGGCCAGGAACAGTCCCGGGTCCTGGGGCCCGAGCTCAGCAGCTATCACCTGGACGGGCTGGAGCCAGCGACACAGTACCGCGTGAGGCTGAGTGTCCTAGGGCCAGCTGGAGAAGGGCCCTCTGCAGAGGTGACTGCGCGCACTGAGTCACCTCGTGTTCCAAGCATTGAACTACGTGTGGTGGACACCTCGATCGACTCGGTGACTTTGGCCTGGACTCCAGTGTCCAGGGCATCCAGCTACATCCTATCCTGGCGGCCACTCAGAGGCCCTGGCCAGGAAGTGCCTGGGTCCCCGCAGACACTTCCAGGGATCTCAAGCTCCCAGCGGGTGACAGGGCTAGAGCCTGGCGTCTCTTACATCTTCTCCCTGACGCCTGTCCTGGATGGTGTGCGGGGTCCTGAGGCATCTGTCACACAGACGCCAGTGTGCCCCCGTGGCCTGGCGGATGTGGTGTTCCTACCACATGCCACTCAAGACAATGCTCACCGTGCGGAGGCTACGAGGAGGGTCCTGGAGCGTCTGGTGTTGGCACTTGGGCCTCTTGGGCCACAGGCAGTTCAGGTTGGCCTGCTGTCTTACAGTCATCGGCCCTCCCCACTGTTCCCACTGAATGGCTCCCATGACCTTGGCATTATCTTGCAAAGGATCCGTGACATGCCCTACATGGACCCAAGTGGGAACAACCTGGGCACAGCCGTGGTCACAGCTCACAGATACATGTTGGCACCAGATGCTCCTGGGCGCCGCCAGCACGTACCAGGGGTGATGGTTCTGCTAGTGGATGAACCCTTGAGAGGTGACATATTCAGCCCCATCCGTGAGGCCCAGGCTTCTGGGCTTAATGTGGTGATGTTGGGAATGGCTGGAGCGGACCCAGAGCAGCTGCGTCGCTTGGCGCCGGGTATGGACTCTGTCCAGACCTTCTTCGCCGTGGATGATGGGCCAAGCCTGGACCAGGCAGTCAGTGGTCTGGCCACAGCCCTGTGTCAGGCATCCTTCACTACTCAGCCCCGGCCAGAGCCCTGCCCAGTGTATTGTCCAAAGGGCCAGAAGGGGGAACCTGGAGAGATGGGCCTGAGAGGACAAGTTGGGCCTCCTGGCGACCCTGGCCTCCCGGGCAGGACCGGTGCTCCCGGCCCCCAGGGGCCCCCTGGAAGTGCCACTGCCAAGGGCGAGAGGGGCTTCCCTGGAGCAGATGGGCGTCCAGGCAGCCCTGGCCGCGCCGGGAATCCTGGGACCCCTGGAGCCCCTGGCCTAAAGGGCTCTCCAGGGTTGCCTGGCCCTCGTGGGGACCCGGGAGAGCGAGGACCTCGAGGCCCAAAGGGGGAGCCGGGGGCTCCCGGACAAGTCATCGGAGGTGAAGGACCTGGGCTTCCTGGGCGGAAAGGGGACCCTGGACCATCGGGCCCCCCTGGACCTCGTGGACCACTGGGGGACCCAGGACCCCGTGGCCCCCCAGGGCTTCCTGGAACAGCCATGAAGGGTGACAAAGGCGATCGTGGGGAGCGGGGTCCCCCTGGACCAGGTGAAGGTGGCATTGCTCCTGGGGAGCCTGGGCTGCCGGGTCTTCCCGGAAGCCCTGGACCCCAAGGCCCCGTTGGCCCCCCTGGAAAGAAAGGAGAAAAAGGTGACTCTGAGGATGGAGCTCCAGGCCTCCCAGGACAACCTGGGTCTCCGGGTGAGCAGGGCCCACGGGGACCTCCTGGAGCTATTGGCCCCAAAGGTGACCGGGGCTTTCCAGGGCCCCTGGGTGAGGCTGGAGAGAAGGGCGAACGTGGACCCCCAGGCCCAGCGGGATCCCGGGGGCTGCCAGGGGTTGCTGGACGTCCTGGAGCCAAGGGTCCTGAAGGGCCACCAGGACCCACTGGCCGCCAAGGAGAGAAGGGGGAGCCTGGTCGCCCTGGGGACCCTGCAGTGGTGGGACCTGCTGTTGCTGGACCCAAAGGAGAAAAGGGAGATGTGGGGCCCGCTGGGCCCAGAGGAGCTACCGGAGTCCAAGGGGAACGGGGCCCACCCGGCTTGGTTCTTCCTGGAGACCCTGGCCCCAAGGGAGACCCTGGAGACCGGGGTCCCATTGGCCTTACTGGCAGAGCAGGACCCCCAGGTGACTCAGGGCCTCCTGGAGAGAAGGGAGACCCTGGGCGGCCTGGCCCCCCAGGACCTGTTGGCCCCCGAGGACGAGATGGTGAAGTTGGAGAGAAAGGTGACGAGGGTCCTCCGGGTGACCCGGGTTTGCCTGGAAAAGCAGGCGAGCGTGGCCTTCGGGGGGCACCTGGAGTTCGGGGGCCTGTGGGTGAAAAGGGAGACCAGGGAGATCCTGGAGAGGATGGACGAAATGGCAGCCCTGGATCATCTGGACCCAAGGGTGACCGTGGGGAGCCGGGTCCCCCAGGACCCCCGGGACGGCTGGTAGACACAGGACCTGGAGCCAGAGAGAAGGGAGAGCCTGGGGACCGCGGACAAGAGGGTCCTCGAGGGCCCAAGGGTGATCCTGGCCTCCCTGGAGCCCCTGGGGAAAGGGGCATTGAAGGGTTTCGGGGACCCCCAGGCCCACAGGGGGACCCAGGTGTCCGAGGCCCAGCAGGAGAAAAGGGTGACCGGGGTCCCCCTGGGCTGGATGGCCGGAGCGGACTGGATGGGAAACCAGGAGCCGCTGGGCCCTCTGGGCCGAATGGTGCTGCAGGCAAAGCTGGGGACCCAGGGAGAGACGGGCTTCCAGGCCTCCGTGGAGAACAGGGCCTCCCTGGCCCCTCTGGTCCCCCTGGATTACCGGGAAAGCCAGGCGAGGATGGCAAACCTGGCCTGAATGGAAAAAACGGAGAACCTGGGGACCCTGGAGAAGACGGGAGGAAGGGAGAGAAAGGAGATTCAGGCGCCTCTGGGAGAGAAGGTCGTGATGGCCCCAAGGGTGAGCGTGGAGCTCCTGGTATCCTTGGACCCCAGGGGCCTCCAGGCCTCCCAGGGCCAGTGGGCCCTCCTGGCCAGGGTTTTCCTGGTGTCCCAGGAGGCACGGGCCCCAAGGGTGACCGTGGGGAGACTGGATCCAAAGGGGAGCAGGGCCTCCCTGGAGAGCGTGGCCTGCGAGGAGAGCCTGGAAGTGTGCCGAATGTGGATCGGTTGCTGGAAACTGCTGGCATCAAGGCATCTGCCCTGCGGGAGATCGTGGAGACCTGGGATGAGAGCTCTGGTAGCTTCCTGCCTGTGCCCGAACGGCGTCGAGGCCCCAAGGGGGACTCAGGCGAACAGGGCCCCCCAGGCAAGGAGGGCCCCATCGGCTTTCCTGGAGAACGCGGGCTGAAGGGCGACCGTGGAGACCCTGGCCCTCAGGGGCCACCTGGTCTGGCCCTTGGGGAGAGGGGCCCCCCCGGGCCTTCCGGCCTTGCCGGGGAGCCTGGAAAGCCTGGTATTCCCGGGCTCCCAGGCAGGGCTGGGGGTGTGGGAGAGGCAGGAAGGCCAGGAGAGAGGGGAGAACGGGGAGAGAAAGGAGAACGTGGAGAACAGGGCAGAGATGGCCCTCCTGGACTCCCTGGAACCCCTGGGCCCCCCGGACCCCCTGGCCCCAAGGTGTCTGTGGATGAGCCAGGTCCTGGACTCTCTGGAGAACAGGGACCCCCTGGACTCAAGGGTGCTAAGGGGGAGCCGGGCAGCAATGGTGACCAAGGTCCCAAAGGAGACAGGGGTGTGCCAGGCATCAAAGGAGACCGGGGAGAGCCTGGACCGAGGGGTCAGGACGGCAACCCGGGTCTACCAGGAGAGCGTGGTATGGCTGGGCCTGAAGGGAAGCCGGGTCTGCAGGGTCCAAGAGGCCCCCCTGGCCCAGTGGGTGGTCATGGAGACCCTGGACCACCTGGTGCCCCGGGTCTTGCTGGCCCTGCAGGACCCCAAGGACCTTCTGGCCTGAAGGGGGAGCCTGGAGAGACAGGACCTCCAGGACGGGGCCTGACTGGACCTACTGGAGCTGTGGGACTTCCTGGACCCCCCGGCCCTTCAGGCCTTGTGGGTCCACAGGGGTCTCCAGGTTTGCCTGGACAAGTGGGGGAGACAGGGAAGCCGGGAGCCCCAGGTCGAGATGGTGCCAGTGGAAAAGATGGAGACAGAGGGAGCCCTGGTGTGCCAGGGTCACCAGGTCTGCCTGGCCCTGTCGGACCTAAAGGAGAACCTGGCCCCACGGGGGCCCCTGGACAGGCTGTGGTCGGGCTCCCTGGAGCAAAGGGAGAGAAGGGAGCCCCTGGAGGCCTTGCTGGAGACCTGGTGGGTGAGCCGGGAGCCAAAGGTGACCGAGGACTGCCAGGGCCGCGAGGCGAGAAGGGTGAAGCTGGCCGTGCAGGGGAGCCCGGAGACCCTGGGGAAGATGGTCAGAAAGGGGCTCCAGGACCCAAAGGTTTCAAGGGTGACCCAGGAGTCGGGGTCCCGGGCTCCCCTGGGCCTCCTGGCCCTCCAGGTGTGAAGGGAGATCTGGGCCTCCCTGGCCTGCCCGGTGCTCCTGGTGTTGTTGGGTTCCCGGGTCAGACAGGCCCTCGAGGAGAGATGGGTCAGCCAGGCCCTAGTGGAGAGCGGGGTCTGGCAGGCCCCCCAGGGAGAGAAGGAATCCCAGGACCCCTGGGGCCACCTGGACCACCGGGGTCAGTGGGACCACCTGGGGCCTCTGGACTCAAAGGAGACAAGGGAGACCCTGGAGTAGGGCTGCCTGGGCCCCGAGGCGAGCGTGGGGAGCCAGGCATCCGGGGTGAAGATGGCCGCCCCGGCCAGGAGGGACCCCGAGGACTCACGGGGCCCCCTGGCAGCAGGGGAGAGCGTGGGGAGAAGGGTGATGTTGGGAGTGCAGGACTAAAGGGTGACAAGGGAGACTCAGCTGTGATCCTGGGGCCTCCAGGCCCACGGGGTGCCAAGGGGGACATGGGTGAACGAGGGCCTCGGGGCTTGGATGGTGACAAAGGACCTCGGGGAGACAATGGGGACCCTGGTGACAAGGGCAGCAAGGGAGAGCCTGGTGACAAGGGCTCAGCCGGGTTGCCAGGACTGCGTGGACTCCTGGGACCCCAGGGTCAACCTGGTGCAGCAGGGATCCCTGGTGACCCGGGATCCCCAGGAAAGGATGGAGTGCCTGGTATCCGAGGAGAAAAAGGAGATGTTGGCTTCATGGGTCCCCGGGGCCTCAAGGGTGAACGGGGAGTGAAGGGAGCCTGTGGCCTTGATGGAGAGAAGGGAGACAAGGGAGAAGCTGGTCCCCCAGGCCGCCCCGGGCTGGCAGGACACAAAGGAGAGATGGGGGAGCCTGGTGTGCCGGGCCAGTCGGGGGCCCCTGGCAAGGAGGGCCTGATCGGTCCCAAGGGTGACCGAGGCTTTGACGGGCAGCCAGGCCCCAAGGGTGACCAGGGCGAGAAAGGGGAGCGGGGAACCCCAGGAATTGGGGGCTTCCCAGGCCCCAGTGGAAATGATGGCTCTGCTGGTCCCCCAGGGCCACCTGGCAGTGTTGGTCCCAGAGGCCCCGAAGGACTTCAGGGCCAGAAGGGTGAGCGAGGTCCCCCCGGAGAGAGAGTGGTGGGGGCTCCTGGGGTCCCTGGAGCTCCTGGCGAGAGAGGGGAGCAGGGGCGGCCAGGGCCTGCCGGTCCTCGAGGCGAGAAGGGAGAAGCTGCACTGACGGAGGATGACATCCGGGGCTTTGTGCGCCAAGAGATGAGTCAGCACTGTGCCTGCCAGGGCCAGTTCATCGCATCTGGATCACGACCCCTCCCTAGTTATGCTGCAGACACTGCCGGCTCCCAGCTCCATGCTGTGCCTGTGCTCCGCGTCTCTCATGCAGAGGAGGAAGAGCGGGTACCCCCTGAGGATGATGAGTACTCTGAATACTCCGAGTATTCTGTGGAGGAGTACCAGGACCCTGAAGCTCCTTGGGATAGTGATGACCCCTGTTCCCTGCCACTGGATGAGGGCTCCTGCACTGCCTACACCCTGCGCTGGTACCATCGGGCTGTGACAGGCAGCACAGAGGCCTGTCACCCTTTTGTCTATGGTGGCTGTGGAGGGAATGCCAACCGTTTTGGGACCCGTGAGGCCTGCGAGCGCCGCTGCCCACCCCGGGTGGTCCAGAGCCAGGGGACAGGTACTGCCCAGGACTGA
ヒトVII型コラーゲンのアミノ酸配列(2944 AA)(配列番号2)
MTLRLLVAALCAGILAEAPRVRAQHRERVTCTRLYAADIVFLLDGSSSIGRSNFREVRSFLEGLVLPFSGAASAQGVRFATVQYSDDPRTEFGLDALGSGGDVIRAIRELSYKGGNTRTGAAILHVADHVFLPQLARPGVPKVCILITDGKSQDLVDTAAQRLKGQGVKLFAVGIKNADPEELKRVASQPTSDFFFFVNDFSILRTLLPLVSRRVCTTAGGVPVTRPPDDSTSAPRDLVLSEPSSQSLRVQWTAASGPVTGYKVQYTPLTGLGQPLPSERQEVNVPAGETSVRLRGLRPLTEYQVTVIALYANSIGEAVSGTARTTALEGPELTIQNTTAHSLLVAWRSVPGATGYRVTWRVLSGGPTQQQELGPGQGSVLLRDLEPGTDYEVTVSTLFGRSVGPATSLMARTDASVEQTLRPVILGPTSILLSWNLVPEARGYRLEWRRETGLEPPQKVVLPSDVTRYQLDGLQPGTEYRLTLYTLLEGHEVATPATVVPTGPELPVSPVTDLQATELPGQRVRVSWSPVPGATQYRIIVRSTQGVERTLVLPGSQTAFDLDDVQAGLSYTVRVSARVGPREGSASVLTVRREPETPLAVPGLRVVVSDATRVRVAWGPVPGASGFRISWSTGSGPESSQTLPPDSTATDITGLQPGTTYQVAVSVLRGREEGPAAVIVARTDPLGPVRTVHVTQASSSSVTITWTRVPGATGYRVSWHSAHGPEKSQLVSGEATVAELDGLEPDTEYTVHVRAHVAGVDGPPASVVVRTAPEPVGRVSRLQILNASSDVLRITWVGVTGATAYRLAWGRSEGGPMRHQILPGNTDSAEIRGLEGGVSYSVRVTALVGDREGTPVSIVVTTPPEAPPALGTLHVVQRGEHSLRLRWEPVPRAQGFLLHWQPEGGQEQSRVLGPELSSYHLDGLEPATQYRVRLSVLGPAGEGPSAEVTARTESPRVPSIELRVVDTSIDSVTLAWTPVSRASSYILSWRPLRGPGQEVPGSPQTLPGISSSQRVTGLEPGVSYIFSLTPVLDGVRGPEASVTQTPVCPRGLADVVFLPHATQDNAHRAEATRRVLERLVLALGPLGPQAVQVGLLSYSHRPSPLFPLNGSHDLGIILQRIRDMPYMDPSGNNLGTAVVTAHRYMLAPDAPGRRQHVPGVMVLLVDEPLRGDIFSPIREAQASGLNVVMLGMAGADPEQLRRLAPGMDSVQTFFAVDDGPSLDQAVSGLATALCQASFTTQPRPEPCPVYCPKGQKGEPGEMGLRGQVGPPGDPGLPGRTGAPGPQGPPGSATAKGERGFPGADGRPGSPGRAGNPGTPGAPGLKGSPGLPGPRGDPGERGPRGPKGEPGAPGQVIGGEGPGLPGRKGDPGPSGPPGPRGPLGDPGPRGPPGLPGTAMKGDKGDRGERGPPGPGEGGIAPGEPGLPGLPGSPGPQGPVGPPGKKGEKGDSEDGAPGLPGQPGSPGEQGPRGPPGAIGPKGDRGFPGPLGEAGEKGERGPPGPAGSRGLPGVAGRPGAKGPEGPPGPTGRQGEKGEPGRPGDPAVVGPAVAGPKGEKGDVGPAGPRGATGVQGERGPPGLVLPGDPGPKGDPGDRGPIGLTGRAGPPGDSGPPGEKGDPGRPGPPGPVGPRGRDGEVGEKGDEGPPGDPGLPGKAGERGLRGAPGVRGPVGEKGDQGDPGEDGRNGSPGSSGPKGDRGEPGPPGPPGRLVDTGPGAREKGEPGDRGQEGPRGPKGDPGLPGAPGERGIEGFRGPPGPQGDPGVRGPAGEKGDRGPPGLDGRSGLDGKPGAAGPSGPNGAAGKAGDPGRDGLPGLRGEQGLPGPSGPPGLPGKPGEDGKPGLNGKNGEPGDPGEDGRKGEKGDSGASGREGRDGPKGERGAPGILGPQGPPGLPGPVGPPGQGFPGVPGGTGPKGDRGETGSKGEQGLPGERGLRGEPGSVPNVDRLLETAGIKASALREIVETWDESSGSFLPVPERRRGPKGDSGEQGPPGKEGPIGFPGERGLKGDRGDPGPQGPPGLALGERGPPGPSGLAGEPGKPGIPGLPGRAGGVGEAGRPGERGERGEKGERGEQGRDGPPGLPGTPGPPGPPGPKVSVDEPGPGLSGEQGPPGLKGAKGEPGSNGDQGPKGDRGVPGIKGDRGEPGPRGQDGNPGLPGERGMAGPEGKPGLQGPRGPPGPVGGHGDPGPPGAPGLAGPAGPQGPSGLKGEPGETGPPGRGLTGPTGAVGLPGPPGPSGLVGPQGSPGLPGQVGETGKPGAPGRDGASGKDGDRGSPGVPGSPGLPGPVGPKGEPGPTGAPGQAVVGLPGAKGEKGAPGGLAGDLVGEPGAKGDRGLPGPRGEKGEAGRAGEPGDPGEDGQKGAPGPKGFKGDPGVGVPGSPGPPGPPGVKGDLGLPGLPGAPGVVGFPGQTGPRGEMGQPGPSGERGLAGPPGREGIPGPLGPPGPPGSVGPPGASGLKGDKGDPGVGLPGPRGERGEPGIRGEDGRPGQEGPRGLTGPPGSRGERGEKGDVGSAGLKGDKGDSAVILGPPGPRGAKGDMGERGPRGLDGDKGPRGDNGDPGDKGSKGEPGDKGSAGLPGLRGLLGPQGQPGAAGIPGDPGSPGKDGVPGIRGEKGDVGFMGPRGLKGERGVKGACGLDGEKGDKGEAGPPGRPGLAGHKGEMGEPGVPGQSGAPGKEGLIGPKGDRGFDGQPGPKGDQGEKGERGTPGIGGFPGPSGNDGSAGPPGPPGSVGPRGPEGLQGQKGERGPPGERVVGAPGVPGAPGERGEQGRPGPAGPRGEKGEAALTEDDIRGFVRQEMSQHCACQGQFIASGSRPLPSYAADTAGSQLHAVPVLRVSHAEEEERVPPEDDEYSEYSEYSVEEYQDPEAPWDSDDPCSLPLDEGSCTAYTLRWYHRAVTGSTEACHPFVYGGCGGNANRFGTREACERRCPPRVVQSQGTGTAQD*
栄養障害型表皮水疱症の治療に用いるための組成物であって、VII型コラーゲンを産生するよう遺伝子改変された栄養障害型表皮水疱症患者の細胞を含み、前記細胞が間葉系幹細胞である、組成物。
[2]
細胞が、COL7A1遺伝子を導入することにより遺伝子改変されている、前記1に記載の組成物。
[3]
細胞のゲノムにCOL7A1遺伝子が導入されている、前記2に記載の組成物。
[4]
COL7A1遺伝子が、配列番号1の核酸配列と70%、80%、85%、90%、95%、96%、97%、98%または99%以上の配列同一性を有する核酸配列を含むか、配列番号2のアミノ酸配列と70%、80%、85%、90%、95%、96%、97%、98%または99%以上の配列同一性を有するアミノ酸配列をコードする核酸配列を含む、前記2または3に記載の組成物。
[5]
間葉系幹細胞が、骨髄由来間葉系幹細胞である、前記1〜4のいずれかに記載の組成物。
[6]
間葉系幹細胞が、組成物に含まれる細胞で最も多い細胞である、前記1〜5のいずれかに記載の組成物。
[7]
間葉系幹細胞が、組成物に含まれる細胞の70%、80%、85%、90%、95%、96%、97%、98または99%以上を占める、前記1〜6のいずれかに記載の組成物。
[8]
組成物が、実質的に間葉系幹細胞以外の細胞を含まない、前記1〜7のいずれかに記載の組成物。
[9]
患部に投与される、前記1〜8のいずれかに記載の組成物。
[10]
水疱内に投与される、前記1〜9のいずれかに記載の組成物。
[11]
ゲノム編集により細胞が遺伝子改変されている、前記1〜10のいずれかに記載の組成物。
[12]
ゲノム編集が、CRISPR/Cas9による、前記11に記載の組成物。
[13]
ウイスルベクターにより細胞が遺伝子改変されている、前記1〜10のいずれかに記載の組成物。
[14]
ウイスルベクターが、レトロウイルスベクターまたはレンチウイルスベクターである、前記13に記載の組成物。
[15]
ウイスルベクターが、レンチウイルスベクターである、前記13または14に記載の組成物。
栄養障害型表皮水疱症の治療に用いるための組成物であって、VII型コラーゲンを産生する細胞を含み、水疱内に投与される、組成物。
[17]
細胞が、VII型コラーゲンを産生するよう遺伝子改変された細胞である、前記16に記載の組成物。
[18]
細胞が、COL7A1遺伝子を導入することにより遺伝子改変されている、前記17に記載の組成物。
[19]
細胞のゲノムにCOL7A1遺伝子が導入されている、前記18に記載の組成物。
[20]
COL7A1遺伝子が、配列番号1の核酸配列と70%、80%、85%、90%、95%、96%、97%、98%または99%以上の配列同一性を有する核酸配列を含むか、配列番号2のアミノ酸配列と70%、80%、85%、90%、95%、96%、97%、98%または99%以上の配列同一性を有するアミノ酸配列をコードする核酸配列を含む、前記18または19に記載の組成物。
[21]
細胞が、栄養障害型表皮水疱症患者の細胞である、前記17〜20のいずれかに記載の組成物。
[22]
細胞が、骨髄から得た細胞である、前記16〜21のいずれかに記載の組成物。
[23]
細胞が、間葉系幹細胞である、前記16〜22のいずれかに記載の組成物。
[24]
間葉系幹細胞が、骨髄由来間葉系幹細胞である、前記23に記載の組成物。
[25]
間葉系幹細胞が、組成物に含まれる細胞で最も多い細胞である、前記23または24に記載の組成物。
[26]
間葉系幹細胞が、組成物に含まれる細胞の70%、80%、85%、90%、95%、96%、97%、98または99%以上を占める、前記23〜25のいずれかに記載の組成物。
[27]
組成物が、実質的に間葉系幹細胞以外の細胞を含まない、前記23〜26のいずれかに記載の組成物。
[28]
ゲノム編集により細胞が遺伝子改変されている、前記17〜27のいずれかに記載の組成物。
[29]
ゲノム編集が、CRISPR/Cas9による、前記28のいずれかに記載の組成物。
[30]
ウイスルベクターにより細胞が遺伝子改変されている、前記17〜27のいずれかに記載の組成物。
[31]
ウイスルベクターが、レトロウイルスベクターまたはレンチウイルスベクターである、前記30に記載の組成物。
[32]
ウイスルベクターが、レンチウイルスベクターである、前記30または31に記載の組成物。
栄養障害型表皮水疱症の治療に使用するための組成物の製造方法であって、
VII型コラーゲンを産生するよう細胞を遺伝子改変すること、および
遺伝子改変された細胞を含む組成物を調製すること
を含む方法。
[34]
栄養障害型表皮水疱症を治療するための方法であって、VII型コラーゲンを産生する細胞を含む組成物を前記患者に投与することを含む方法。
[35]
細胞が、VII型コラーゲンを産生するよう遺伝子改変された細胞である、前記34に記載の方法。
[36]
細胞が、COL7A1遺伝子を導入することにより遺伝子改変されている、前記35に記載の方法。
[37]
細胞のゲノムにCOL7A1遺伝子が導入されている、前記36に記載の方法。
[38]
COL7A1遺伝子が、配列番号1の核酸配列と70%、80%、85%、90%、95%、96%、97%、98%または99%以上の配列同一性を有する核酸配列を含むか、配列番号2のアミノ酸配列と70%、80%、85%、90%、95%、96%、97%、98%または99%以上の配列同一性を有するアミノ酸配列をコードする核酸配列を含む、前記36または37に記載の方法。
[39]
ゲノム編集により細胞が遺伝子改変されている、前記35〜38のいずれかに記載の方法。
[40]
ゲノム編集が、CRISPR/Cas9による、前記39に記載の方法。
[41]
ウイスルベクターにより細胞が遺伝子改変されている、前記35〜38のいずれかに記載の方法。
[42]
ウイスルベクターが、レトロウイルスベクターまたはレンチウイルスベクターである、前記41に記載の方法。
[43]
ウイスルベクターが、レンチウイルスベクターである、前記41または42に記載の方法。
[44]
患者への投与に先立ち、VII型コラーゲンを産生するよう細胞を遺伝子改変することをさらに含む、前記35〜43のいずれかに記載の方法。
[45]
COL7A1遺伝子を導入することにより細胞を遺伝子改変する、前記33または44に記載の方法。
[46]
細胞のゲノムにCOL7A1遺伝子を導入する、前記45に記載の方法。
[47]
COL7A1遺伝子が、配列番号1の核酸配列と70%、80%、85%、90%、95%、96%、97%、98%または99%以上の配列同一性を有する核酸配列を含むか、配列番号2のアミノ酸配列と70%、80%、85%、90%、95%、96%、97%、98%または99%以上の配列同一性を有するアミノ酸配列をコードする核酸配列を含む、前記45または46に記載の方法。
[48]
ゲノム編集により細胞を遺伝子改変する、前記33および44〜47のいずれかに記載の方法。
[49]
ゲノム編集が、CRISPR/Cas9による、前記48に記載の方法。
[50]
ウイスルベクターにより細胞を遺伝子改変する、前記33および44〜47のいずれかに記載の方法。
[51]
ウイスルベクターが、レトロウイルスベクターまたはレンチウイルスベクターである、前記50に記載の方法。
[52]
ウイスルベクターが、レンチウイルスベクターである、前記50または51に記載の方法。
[53]
細胞が、栄養障害型表皮水疱症患者の細胞である、前記33〜52のいずれかに記載の方法。
[54]
遺伝子改変に先立ち、栄養障害型表皮水疱症患者から細胞を得ることをさらに含む、前記33および44〜53のいずれかに記載の方法。
[55]
細胞が、骨髄から得た細胞である、前記33〜54のいずれかに記載の方法。
[56]
細胞が、間葉系幹細胞である、前記33〜55のいずれかに記載の方法。
[57]
間葉系幹細胞が、骨髄由来間葉系幹細胞である、前記56に記載の方法。
[58]
間葉系幹細胞が、組成物に含まれる細胞で最も多い細胞である、前記56または57に記載の方法。
[59]
間葉系幹細胞が、組成物に含まれる細胞の70%、80%、85%、90%、95%、96%、97%、98または99%以上を占める、前記56〜58のいずれかに記載の方法。
[60]
組成物が、実質的に間葉系幹細胞以外の細胞を含まない、前記56〜59のいずれかに記載の方法。
[61]
組成物が、患部に投与される、前記34〜60のいずれかに記載の方法。
[62]
組成物が、水疱内に投与される、前記34〜61のいずれかに記載の方法。
栄養障害型表皮水疱症を治療するための医薬の製造のための、VII型コラーゲンを産生するよう遺伝子改変された栄養障害型表皮水疱症患者の細胞を含む組成物の使用であって、前記細胞が間葉系幹細胞を含む、使用。
[64]
栄養障害型表皮水疱症を治療するための医薬の製造のための、VII型コラーゲンを産生する細胞を含む組成物の使用であって、組成物が水疱内に投与される、使用。
栄養障害型表皮水疱症を治療するための、VII型コラーゲンを産生するよう遺伝子改変された栄養障害型表皮水疱症患者の細胞を含む組成物の使用であって、前記細胞が間葉系幹細胞を含む、使用。
[66]
栄養障害型表皮水疱症を治療するための、VII型コラーゲンを産生する細胞を含む組成物の使用であって、組成物が水疱内に投与される、使用。
配列番号3〜5のいずれかの配列またはこれに相補的な配列を含む、gRNA。
[68]
前記67のgRNAをコードする核酸配列を含むベクター。
ヒトゲノム内のAAVS1(Adeno-associated virus integration site 1)領域においてCRISPR-Cas9システムによる切断効率が良好な位置を選択するため、3種類のsgRNAを作製した。AAVS1領域は、遺伝子導入による影響を受けにくい安全領域(セーフ・ハーバー/safe harbor)である。CRISPR-Cas9システムは「NGG」の塩基配列を認識し、その3塩基上流を切断することから、末端に「GG」が配置される領域を選択し、「NGG」の上流20塩基の配列を含むsgRNA(sgAAVS1-#1〜#3)を設計した(図1、上;表1)。
AAVS1領域へのCOL7A1遺伝子の導入のため、CAGプロモーターの制御下にCOL7A1遺伝子を発現するプラスミドを設計した(図2)。COL7A1 cDNAは、Flexi ORF sequence-verified clone (Promega, Madison, WI, USA)より得た。COL7A1 cDNAをpENTR1Aプラスミド(ThermoFisher Scientific)にサブクローニングし、pENTR1A-COL7A1を作製し、LRリコンビナーゼ (ThermoFisher Scientific) とpAAVS-P-CAG-DEST (Addgene plasmid # 80490)およびpENTR1A-COL7A1とを用いて、pAAVS-P-CAG-COL7A1を得た。
表皮水疱症モデルマウスに対する遺伝子改変MSCの投与によるVII型コラーゲンの発現を検討した。水疱形成を示すCol7A1遺伝子ノックアウトマウス(Col7a1-/-)の新生児の全層皮膚を切除し、免疫不全マウス(NOD-SCID)の背部に移植した。移植の1週間後、前記2の遺伝子改変MSCを、0.1〜1.0 × 106細胞で皮下または皮内へ注入した(図6)。水疱内注入では、移植直後に皮膚表面をつまみ、かつ擦ることで水疱を形成させ、直ちに表皮下の空間に0.1〜1.0 × 106細胞の遺伝子改変MSCを注入した。それぞれ注入の4週間後、移植皮膚を切除し、抗VII型コラーゲン抗体(Sigma Aldrich, St. Louis, MO, USA)を用いた免疫染色により、VII型コラーゲンの発現を評価した。
Claims (16)
- 栄養障害型表皮水疱症の治療に用いるための組成物であって、VII型コラーゲンを産生するよう遺伝子改変された栄養障害型表皮水疱症患者の細胞を含み、前記細胞が間葉系幹細胞である、組成物。
- 細胞が、COL7A1遺伝子を導入することにより遺伝子改変されている、請求項1に記載の組成物。
- COL7A1遺伝子が、配列番号1の核酸配列と90%以上の配列同一性を有する核酸配列を含むか、配列番号2のアミノ酸配列と90%以上の配列同一性を有するアミノ酸配列をコードする核酸配列を含む、請求項2に記載の組成物。
- 間葉系幹細胞が、骨髄由来間葉系幹細胞である、請求項1〜3のいずれかに記載の組成物。
- 間葉系幹細胞が、組成物に含まれる細胞で最も多い細胞である、請求項1〜4のいずれかに記載の組成物。
- 水疱内に投与される、請求項1〜5のいずれかに記載の組成物。
- 栄養障害型表皮水疱症の治療に用いるための組成物であって、VII型コラーゲンを産生する細胞を含み、水疱内に投与される、組成物。
- 細胞が、VII型コラーゲンを産生するよう遺伝子改変された細胞である、請求項7に記載の組成物。
- 細胞が、COL7A1遺伝子を導入することにより遺伝子改変されている、請求項8に記載の組成物。
- COL7A1遺伝子が、配列番号1の核酸配列と90%以上の配列同一性を有する核酸配列を含むか、配列番号2のアミノ酸配列と90%以上の配列同一性を有するアミノ酸配列をコードする核酸配列を含む、請求項9に記載の組成物。
- 細胞が、栄養障害型表皮水疱症患者の細胞である、前記8〜10のいずれかに記載の組成物。
- 細胞が、骨髄から得た細胞である、請求項7〜11のいずれかに記載の組成物。
- 細胞が、間葉系幹細胞である、請求項7〜12のいずれかに記載の組成物。
- 間葉系幹細胞が、骨髄由来間葉系幹細胞である、請求項13に記載の組成物。
- 間葉系幹細胞が、組成物に含まれる細胞で最も多い細胞である、前記13または14に記載の組成物。
- 配列番号3〜5のいずれかの配列またはこれに相補的な配列を含む、gRNA。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2019007201 | 2019-01-18 | ||
JP2019007201 | 2019-01-18 | ||
PCT/JP2020/001433 WO2020149395A1 (ja) | 2019-01-18 | 2020-01-17 | 栄養障害型表皮水疱症治療薬 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2020149395A1 true JPWO2020149395A1 (ja) | 2021-11-25 |
JPWO2020149395A5 JPWO2020149395A5 (ja) | 2023-01-26 |
Family
ID=71613343
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2020566497A Pending JPWO2020149395A1 (ja) | 2019-01-18 | 2020-01-17 | 栄養障害型表皮水疱症治療薬 |
Country Status (7)
Country | Link |
---|---|
US (1) | US20220088083A1 (ja) |
EP (1) | EP3912644A4 (ja) |
JP (1) | JPWO2020149395A1 (ja) |
KR (1) | KR20210116531A (ja) |
CN (1) | CN113646006A (ja) |
AU (1) | AU2020209687A1 (ja) |
WO (1) | WO2020149395A1 (ja) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2020459825A1 (en) * | 2020-07-22 | 2023-03-16 | Osaka University | Therapeutic agent for dystrophic epidermolysis bullosa |
WO2023140349A1 (ja) * | 2022-01-21 | 2023-07-27 | 国立大学法人大阪大学 | 細胞シート |
CN116555349A (zh) * | 2023-01-09 | 2023-08-08 | 中吉智药(南京)生物技术有限公司 | 一种腺相关病毒载体及其构建方法与应用 |
Family Cites Families (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090263900A1 (en) * | 2008-04-14 | 2009-10-22 | Sangamo Biosciences, Inc. | Linear donor constructs for targeted integration |
BR112014027813A2 (pt) * | 2012-05-07 | 2017-08-08 | Dow Agrosciences Llc | métodos e composições para integração de transgenes direcionada mediada por nuclease |
US20180104186A1 (en) * | 2013-03-13 | 2018-04-19 | University Of Miami | Methods and compositions for the treatment of epidermolysis bullosa |
CN105683376A (zh) * | 2013-05-15 | 2016-06-15 | 桑格摩生物科学股份有限公司 | 用于治疗遗传病状的方法和组合物 |
AU2017205925B2 (en) | 2016-01-04 | 2020-10-15 | The Board Of Trustees Of The Leland Stanford Junior University | Gene therapy for recessive dystrophic Epidermolysis Bullosa using genetically corrected autologous keratinocytes |
US20190192636A1 (en) * | 2016-03-18 | 2019-06-27 | Intrexon Corporation | Compositions and methods for treatment of type viii collagen deficiencies |
DK3377637T3 (da) * | 2016-04-08 | 2020-05-18 | Krystal Biotech Inc | Sammensætninger til anvendelse i fremgangsmåder til behandling af sår, lidelser og sygdomme i huden |
WO2018007871A1 (en) * | 2016-07-08 | 2018-01-11 | Crispr Therapeutics Ag | Materials and methods for treatment of transthyretin amyloidosis |
US10648002B2 (en) * | 2016-11-22 | 2020-05-12 | Regents Of The University Of Minnesota | Method for correcting a genetic sequence |
US20190365929A1 (en) | 2017-02-22 | 2019-12-05 | Crispr Therapeutics Ag | Materials and methods for treatment of dystrophic epidermolysis bullosa (deb) and other collagen type vii alpha 1 chain (col7a1) gene related conditions or disorders |
RU2019134382A (ru) * | 2017-04-07 | 2021-05-07 | Стемрим Инк. | Лекарственное средство для лечения фиброзного заболевания |
WO2018235834A1 (ja) * | 2017-06-19 | 2018-12-27 | 国立大学法人北海道大学 | 表皮水疱症の治療剤 |
JP2019007201A (ja) | 2017-06-23 | 2019-01-17 | システム計測株式会社 | 免震構造 |
CN107904208B (zh) * | 2017-12-25 | 2019-11-01 | 云舟生物科技(广州)有限公司 | 细胞表型研究用的细胞克隆及其筛选方法和应用 |
CA3091688A1 (en) * | 2018-04-27 | 2019-10-31 | Seattle Children's Hospital (dba Seattle Children's Research Institute) | Expression of foxp3 in edited cd34+ cells |
-
2020
- 2020-01-17 US US17/423,285 patent/US20220088083A1/en active Pending
- 2020-01-17 CN CN202080022396.3A patent/CN113646006A/zh active Pending
- 2020-01-17 KR KR1020217025601A patent/KR20210116531A/ko unknown
- 2020-01-17 AU AU2020209687A patent/AU2020209687A1/en active Pending
- 2020-01-17 EP EP20741062.2A patent/EP3912644A4/en active Pending
- 2020-01-17 WO PCT/JP2020/001433 patent/WO2020149395A1/ja unknown
- 2020-01-17 JP JP2020566497A patent/JPWO2020149395A1/ja active Pending
Also Published As
Publication number | Publication date |
---|---|
EP3912644A4 (en) | 2022-09-28 |
AU2020209687A1 (en) | 2021-07-29 |
WO2020149395A1 (ja) | 2020-07-23 |
KR20210116531A (ko) | 2021-09-27 |
EP3912644A1 (en) | 2021-11-24 |
US20220088083A1 (en) | 2022-03-24 |
CN113646006A (zh) | 2021-11-12 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7158427B2 (ja) | ヘモグロビン異常症を処置するためのグロビン遺伝子療法 | |
JP7233375B2 (ja) | リソソーム障害を治療する方法 | |
WO2020149395A1 (ja) | 栄養障害型表皮水疱症治療薬 | |
JP2018516556A (ja) | ヌクレアーゼ介在性遺伝子発現調節 | |
US11753654B2 (en) | Globin gene therapy for treating hemoglobinopathies | |
JP2024506751A (ja) | 遺伝子サイレンシング | |
JP2023033551A (ja) | ヘモグロビン異常症を治療するためのベクターおよび組成物 | |
WO2019060297A1 (en) | MITOCHONDRIAL REJUVENATION AS A TREATMENT FOR AGE-RELATED AND NEURODEGENERATIVE CONDITIONS | |
US20240139254A1 (en) | Mesenchymal stem cell having oxidative stress resistance, preparation method therefor, and use thereof | |
WO2022018884A1 (ja) | 栄養障害型表皮水疱症の治療薬 | |
JP2019514422A (ja) | Pklrの遺伝子発現増強のための組成物および方法 | |
JP2022512674A (ja) | 人工トランス活性化因子による選択 | |
TALENS et al. | 20 20 A. Aslan and SA Yuka | |
Gupta et al. | Genome‐Editing Applications in Stem Cell Engineering and Regenerative Medicine | |
WO2022198126A9 (en) | Methods for generating mechanically-responsive cells and uses thereof | |
KR20230131816A (ko) | 저면역원성 줄기세포, 줄기세포로부터 분화되거나 유래된 저면역원성 세포및 이의 제조방법 | |
JP2024516459A (ja) | 遺伝子治療のための新規プロモータ配列 | |
Papanikolaou et al. | Genetic manipulation of stem cells |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20230116 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20230116 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20231205 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20240129 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20240311 |