HRP20220482T1 - Postupci in vitro diferencijacije neurona dopamina srednjeg mozga (mda) - Google Patents
Postupci in vitro diferencijacije neurona dopamina srednjeg mozga (mda) Download PDFInfo
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- HRP20220482T1 HRP20220482T1 HRP20220482TT HRP20220482T HRP20220482T1 HR P20220482 T1 HRP20220482 T1 HR P20220482T1 HR P20220482T T HRP20220482T T HR P20220482TT HR P20220482 T HRP20220482 T HR P20220482T HR P20220482 T1 HRP20220482 T1 HR P20220482T1
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- 238000000034 method Methods 0.000 title claims 17
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 title claims 6
- 229960003638 dopamine Drugs 0.000 title claims 3
- 210000002569 neuron Anatomy 0.000 title claims 3
- 230000004069 differentiation Effects 0.000 title claims 2
- 238000000338 in vitro Methods 0.000 title claims 2
- 210000001259 mesencephalon Anatomy 0.000 title claims 2
- 210000004027 cell Anatomy 0.000 claims 23
- 230000011664 signaling Effects 0.000 claims 19
- 239000012190 activator Substances 0.000 claims 12
- 102000003693 Hedgehog Proteins Human genes 0.000 claims 11
- 108090000031 Hedgehog Proteins Proteins 0.000 claims 11
- 230000004156 Wnt signaling pathway Effects 0.000 claims 9
- 239000003112 inhibitor Substances 0.000 claims 9
- 210000001671 embryonic stem cell Anatomy 0.000 claims 6
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims 4
- 102000007350 Bone Morphogenetic Proteins Human genes 0.000 claims 4
- 108010007726 Bone Morphogenetic Proteins Proteins 0.000 claims 4
- 108010087745 Hepatocyte Nuclear Factor 3-beta Proteins 0.000 claims 4
- 102100029284 Hepatocyte nuclear factor 3-beta Human genes 0.000 claims 4
- 102100040290 LIM homeobox transcription factor 1-alpha Human genes 0.000 claims 4
- 101710178355 LIM homeobox transcription factor 1-alpha Proteins 0.000 claims 4
- 241000288906 Primates Species 0.000 claims 4
- 241000283984 Rodentia Species 0.000 claims 4
- 102000004887 Transforming Growth Factor beta Human genes 0.000 claims 4
- 108090001012 Transforming Growth Factor beta Proteins 0.000 claims 4
- 229940112869 bone morphogenetic protein Drugs 0.000 claims 4
- 210000004263 induced pluripotent stem cell Anatomy 0.000 claims 3
- 210000001778 pluripotent stem cell Anatomy 0.000 claims 3
- CDOVNWNANFFLFJ-UHFFFAOYSA-N 4-[6-[4-(1-piperazinyl)phenyl]-3-pyrazolo[1,5-a]pyrimidinyl]quinoline Chemical compound C1CNCCN1C1=CC=C(C2=CN3N=CC(=C3N=C2)C=2C3=CC=CC=C3N=CC=2)C=C1 CDOVNWNANFFLFJ-UHFFFAOYSA-N 0.000 claims 2
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 claims 2
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 claims 2
- IVOMOUWHDPKRLL-KQYNXXCUSA-N Cyclic adenosine monophosphate Chemical compound C([C@H]1O2)OP(O)(=O)O[C@H]1[C@@H](O)[C@@H]2N1C(N=CN=C2N)=C2N=C1 IVOMOUWHDPKRLL-KQYNXXCUSA-N 0.000 claims 2
- FHYUGAJXYORMHI-UHFFFAOYSA-N SB 431542 Chemical compound C1=CC(C(=O)N)=CC=C1C1=NC(C=2C=C3OCOC3=CC=2)=C(C=2N=CC=CC=2)N1 FHYUGAJXYORMHI-UHFFFAOYSA-N 0.000 claims 2
- 108091005735 TGF-beta receptors Proteins 0.000 claims 2
- 102000016715 Transforming Growth Factor beta Receptors Human genes 0.000 claims 2
- 108091000117 Tyrosine 3-Monooxygenase Proteins 0.000 claims 2
- 102000048218 Tyrosine 3-monooxygenases Human genes 0.000 claims 2
- 239000000556 agonist Substances 0.000 claims 2
- 229960005070 ascorbic acid Drugs 0.000 claims 2
- 235000010323 ascorbic acid Nutrition 0.000 claims 2
- 239000011668 ascorbic acid Substances 0.000 claims 2
- 229940077737 brain-derived neurotrophic factor Drugs 0.000 claims 2
- AQGNHMOJWBZFQQ-UHFFFAOYSA-N CT 99021 Chemical compound CC1=CNC(C=2C(=NC(NCCNC=3N=CC(=CC=3)C#N)=NC=2)C=2C(=CC(Cl)=CC=2)Cl)=N1 AQGNHMOJWBZFQQ-UHFFFAOYSA-N 0.000 claims 1
- DWJXYEABWRJFSP-XOBRGWDASA-N DAPT Chemical compound N([C@@H](C)C(=O)N[C@H](C(=O)OC(C)(C)C)C=1C=CC=CC=1)C(=O)CC1=CC(F)=CC(F)=C1 DWJXYEABWRJFSP-XOBRGWDASA-N 0.000 claims 1
- 102000034615 Glial cell line-derived neurotrophic factor Human genes 0.000 claims 1
- 108091010837 Glial cell line-derived neurotrophic factor Proteins 0.000 claims 1
- 101000668058 Infectious salmon anemia virus (isolate Atlantic salmon/Norway/810/9/99) RNA-directed RNA polymerase catalytic subunit Proteins 0.000 claims 1
- 108010025020 Nerve Growth Factor Proteins 0.000 claims 1
- 102000007072 Nerve Growth Factors Human genes 0.000 claims 1
- 102000007399 Nuclear hormone receptor Human genes 0.000 claims 1
- 108020005497 Nuclear hormone receptor Proteins 0.000 claims 1
- 102000056172 Transforming growth factor beta-3 Human genes 0.000 claims 1
- 108090000097 Transforming growth factor beta-3 Proteins 0.000 claims 1
- 102000013814 Wnt Human genes 0.000 claims 1
- 108050003627 Wnt Proteins 0.000 claims 1
- 102000052547 Wnt-1 Human genes 0.000 claims 1
- 108700020987 Wnt-1 Proteins 0.000 claims 1
- 102000044880 Wnt3A Human genes 0.000 claims 1
- 108700013515 Wnt3A Proteins 0.000 claims 1
- 238000011977 dual antiplatelet therapy Methods 0.000 claims 1
- 230000002518 glial effect Effects 0.000 claims 1
- 230000035800 maturation Effects 0.000 claims 1
- 239000003900 neurotrophic factor Substances 0.000 claims 1
- 108020004017 nuclear receptors Proteins 0.000 claims 1
- 239000002243 precursor Substances 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 claims 1
- FYBHCRQFSFYWPY-UHFFFAOYSA-N purmorphamine Chemical compound C1CCCCC1N1C2=NC(OC=3C4=CC=CC=C4C=CC=3)=NC(NC=3C=CC(=CC=3)N3CCOCC3)=C2N=C1 FYBHCRQFSFYWPY-UHFFFAOYSA-N 0.000 claims 1
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- 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
- C12N5/0618—Cells of the nervous system
- C12N5/0619—Neurons
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- 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
- A61K35/30—Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue
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- A61P25/00—Drugs for disorders of the nervous system
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- A61P25/16—Anti-Parkinson drugs
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- A61P25/00—Drugs for disorders of the nervous system
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- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/13—Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
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- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
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- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
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- C12N2506/02—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from embryonic cells
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Claims (14)
1. Postupak in vitro diferencijacije pluripotentnih matičnih stanica obuhvaća:
(a) dovođenje u kontakt više pluripotentnih matičnih stanica sa najmanje jednim inhibitorom TGFβ/aktivin-nodalnog signaliziranja; i
(b) dovođenje u kontakt stanica sa najmanje jednim aktivatorom signaliziranja zvučnog ježa (SHH) i najmanje jednim aktivatorom signaliziranja wnt-signalnog puta (Wnt),
pri čemu je koncentracija barem jednog aktivatora signaliziranja Wnt-signalnog puta povećana (i) između 2 dana i 6 dana od njegovog početnog kontakta sa stanicama i (ii) za između 250% i 1800% njegove početne koncentracije dovedene u kontakt sa stanicama, da bi se dobila populacija diferenciranih stanica koje izražavaju forkhead box protein A2 (FOXA2) i LIM homeobox transkripcioni faktor 1 alfa (LMX1A).
2. Postupak iz patentnog zahtjeva 1, još obuhvaća dovođenje u kontakt stanica sa najmanje jednim inhibitorom signaliziranja koštano morfogenetskog proteina (BMP) i Small Mothers Against Decapentaplegic (SMAD) signaliziranja.
3. Postupak iz patentnog zahtjeva 2, pri čemu se stanice dovode u kontakt sa najmanje jednim inhibitorom TGFβ/aktivin-nodalnog signaliziranja, najmanje jednim inhibitorom BMP/SMAD signaliziranja, i najmanje jednim aktivatorom signaliziranja zvučnog ježa (SHH) tokom između 4 dana i 10 dana, ili i do 7 dana, ili tokom najmanje 7 dana.
4. Postupak iz patentnog zahtjeva 2, pri čemu se stanice dovode u kontakt sa najmanje jednim aktivatorom signaliziranja Wnt signalnog puta tokom između 8 i 15 dana, ili tokom ili do 12 dana, ili tokom najmanje 12 dana.
5. Postupak iz bilo kojeg od patentnih zahtjeva 1-4, pri čemu koncentracija najmanje jednog aktivatora signaliziranja Wnt signalnog puta jeste povećana 4 dana od njegovog početnog kontakta sa stanicama.
6. Postupak iz bilo kojeg od patentnih zahtjeva 1-5, pri čemu povećanje koncentracije najmanje jednog aktivatora signaliziranja Wnt signalnog puta je 400% do 1450% od prvobitne koncentracije najmanje jednog aktivatora signaliziranja wnt signalnog puta dovedene u kontakt sa stanicama, ili 700% do 1050% prvobitne koncentracije najmanje jednog aktivatora signaliziranja wnt signalnog puta dovedenog u kontakt s tim stanicama.
7. Postupak iz bilo kojeg od patentnih zahtjeva 1-6, pri čemu povećanje koncentracije najmanje jednog aktivatora signaliziranja Wnt signalnog puta je povećanje do koncentracije od između 3 µM i 10 µM, ili 3 µM, ili 7,5 µM.
8. Postupak iz bilo kojeg od patentnih zahtjeva 1-7, pri čemu te diferencirane stanice izražavaju jednu ili više tirozinskih hidroksilaza (TH), utisnuti engrailed-1 (EN-1), i sa nuklearnim receptorom srodan-1 protein (NURR1), opcionalno pri čemu te diferencirane stanice ne izražavaju detektibilne nivoe familije uparenog boks proteina (PAX6) i/ili Ki67.
9. Postupak iz bilo kojeg od patentnih zahtjeva 1-8, pri čemu taj postupak još obuhvaća izlaganje populacije diferenciranih stanica uvjetima koji pogoduju sazrijevanju stanica u neuronima dopamina, opcionalno pri čemu ti uvjeti obuhvaćaju dovođenje u kontakt stanica sa neurotrofnim moždanim faktorom (BDNF), neurotrofnim glijalnim faktorom (GDNF), cikličnim adenozin monofosfatom (cAMP), transformišućim faktorom rasta beta 3 (TGFβ3), askorbinskom kiselinom (AA), i/ili DAPT.
10. Postupak iz bilo kojeg od patentnih zahtjeva 1-9, pri čemu se te pluripotentne stanice biraju iz grupe koja obuhvaća humane ne-embrione matične stanice, ne-embrione matične stanice primata, ne-embrione matične stanice glodavaca, humane embrione matične stanice, embrione matične stanice primata, embrione matične stanice glodara, pluripotentne matične stanice inducirane iz čovjeka, pluripotentne matične stanice inducirane iz primata, pluripotentne matične stanice inducirane iz glodavca, rekombianantne pluripotentne stanice čovjeka, rekombinantne pluripotentne stanice primata, i rekombinantne pluripotentne stanice glodavca.
11. Postupak iz bilo kojeg od patentnih zahtjeva 2-10, pri čemu
(a) najmanje jedan inhibitor TGFβ/aktivin-nodalnog signaliziranja obuhvaća inhibitor TGFβ receptora, opcionalno pri čemu taj inhibitor TGFβ receptora obuhvaća 4-[4-(1,3-benzodioksol-5-il)-5-(2-piridinil)-1H-imidazol-2-il] benzamid (SB431542); i/ili
(b) najmanje jedan inhibitor BMP i SMAD signaliziranja obuhvaća 4-(6-(4-(piperazin-1- il)fenil)pirazolo[1,5-a]pirimidin-3-il)hinolin (LDN193189), nogin, kombinaciju gore navedenih; i/ili
(c) najmanje jedan aktivator SHH signaliziranja obuhvaća SHH protein, zaglađeni (SMO) agonist, ili kombinaciju gore navedenih, opcionalno (i) pri čemu taj SHH protein obuhvaća rekombinantni SHH, prečišćeni SHH, ili kombinaciju gore navedenih, i/ili (ii) pri čemu taj rekombinantni SHH obuhvaća SHH C25II, i/ili (iii) pri čemu zaglađeni (SMO) agonist obuhvaća purmorfamin; i/ili
(d) najmanje jedan aktivator signaliziranja Wnt signalnog puta obuhvaća CHIR99021, Wnt3A, Wnt1, ili kombinaciju gore navedenih.
12. Postupak iz bilo kojeg od patentnih zahtjeva 1-11, pri čemu su diferencirane stanice koje izražavaju FOXA2 i LMX1A neuroni dopamina srednjeg mozga ili njihovi prekursori.
13. Postupak iz bilo kojeg od patentnih zahtjeva 1-12, pri čemu su pluripotentne matične stanice diferencirane u diferencirane stanice koje izražavaju FOXA2 i LMX1A ne kasnije od između 22 dana i 27 dana od njihovog početnog kontakta sa najmanje jednim inhibitorom TGFβ/aktivin-nodalnog signaliziranja.
14. Postupak iz bilo kojeg od patentnih zahtjeva 1-13, pri čemu te diferencirane stanice izražavaju detektibilni nivo CD142, opcionalno pri čemu taj postupak još obuhvaća odabir populacije stanica koje izražavaju CD142.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201562169444P | 2015-06-01 | 2015-06-01 | |
| US201562169379P | 2015-06-01 | 2015-06-01 | |
| EP16804350.3A EP3303564B1 (en) | 2015-06-01 | 2016-06-01 | Methods of in vitro differentiation of midbrain dopamine (mda) neurons |
| PCT/US2016/035312 WO2016196661A1 (en) | 2015-06-01 | 2016-06-01 | Methods of in vitro differentiation of midbrain dopamine (mda) neurons |
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| Publication Number | Publication Date |
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| HRP20220482T1 true HRP20220482T1 (hr) | 2022-07-08 |
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| HRP20220482TT HRP20220482T1 (hr) | 2015-06-01 | 2016-06-01 | Postupci in vitro diferencijacije neurona dopamina srednjeg mozga (mda) |
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| Country | Link |
|---|---|
| US (2) | US10858625B2 (hr) |
| EP (3) | EP4070803A1 (hr) |
| JP (4) | JP6974180B2 (hr) |
| KR (3) | KR102679619B1 (hr) |
| AU (2) | AU2016270793B2 (hr) |
| CA (1) | CA2987617A1 (hr) |
| DK (1) | DK3303564T3 (hr) |
| ES (1) | ES2910032T3 (hr) |
| HR (1) | HRP20220482T1 (hr) |
| HU (1) | HUE058258T2 (hr) |
| IL (3) | IL313799A (hr) |
| LT (1) | LT3303564T (hr) |
| PL (1) | PL3303564T3 (hr) |
| PT (1) | PT3303564T (hr) |
| RS (1) | RS63157B1 (hr) |
| SI (1) | SI3303564T1 (hr) |
| WO (1) | WO2016196661A1 (hr) |
Families Citing this family (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| HRP20220482T1 (hr) * | 2015-06-01 | 2022-07-08 | Memorial Sloan Kettering Cancer Center | Postupci in vitro diferencijacije neurona dopamina srednjeg mozga (mda) |
| WO2017136834A1 (en) * | 2016-02-05 | 2017-08-10 | Memorial Sloan-Ketering Cancer Center | Methods of differentiating stem cell-derived ectodermal lineage precursors |
| DK3500664T3 (da) | 2016-08-16 | 2021-12-06 | Fujifilm Cellular Dynamics Inc | Fremgangsmåde til differentiering af pluripotente celler |
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