Reprogramming fibroblasts into cardiomyocytes
Compositions and methods are described herein for chemically inducing cells that express a single pluripotency transcription factor to change their differentiation state and become cardiac cells, cardiac progenitor cells, cardiomyocytes, or a combination thereof.
1. A method of converting a fibroblast into a cardiomyocyte or cardiomyocyte-like cell comprising:
contacting a fibroblast with an agent that induces Oct4 polypeptide expression; and
contacting a fibroblasts with at least four of the following agents: a WNT agonist, a GSK3 inhibitor, a TGF-beta inhibitor, an epigenetic modifier, or an adenylyl cyclase agonist.
2. The method of claim 1 , wherein the fibroblast is contacted with an agent that induces Oct4 polypeptide expression and with a TGF-beta inhibitor, a GSK3 inhibitor, an epigenetic modifier, and an adenylyl cyclase agonist.
3. The method of claim 1 , wherein the fibroblast is contacted with an agent that induces Oct4 polypeptide expression and with an ALK4/5/7 inhibitor, a GSK3 inhibitor, a LSD1/KDM1 inhibitor and an adenylyl cyclase agonist.
4. A method of converting a mammalian fibroblast into a cardiomyocyte or cardiomyocyte-like cell, comprising:
contacting a mammalian fibroblast with an agent that induces Oct4 polypeptide expression; and
contacting the mammalian fibroblast with at least four of the following agents: a WNT agonist, a GSK3 inhibitor, a TGF-beta inhibitor, an epigenetic modifier, or an adenylyl cyclase agonist.
5. The method of claim 4 , wherein the mammalian fibroblast is contacted with an agent that induces Oct4 polypeptide expression and with a TGF-beta inhibitor, a GSK3 inhibitor, an epigenetic modifier, and an adenylyl cyclase agonist.
6. The method of claim 4 , wherein the mammalian fibroblast is contacted with an agent that induces Oct4 polypeptide expression and with an ALK4/5/7 inhibitor, a GSK3 inhibitor, a LSD1/KDM1 inhibitor and an adenylyl cyclase agonist.
7. The method of claim 4 , wherein the mammalian fibroblast is contacted with a vector for expression of Oct4 polypeptides.
8. The method of claim 4 , wherein the mammalian fibroblast is contacted with Oct4 RNA molecules.
9. The method of claim 4 , wherein the mammalian fibroblast comprises an introduced RNA comprising an open reading frame (ORF) for the Oct4 polypeptide flanked by a 5′ untranslated region (UTR) containing a translational initiation signal.
10. The method of claim 4 , wherein the mammalian fibroblast expresses an Oct4 polypeptide from a heterologous promoter.
11. The method of claim 10 , wherein a heterologous Oct4 nucleic acid is not integrated into the mammalian fibroblast's genome.
12. The method of claim 4 , wherein the mammalian fibroblast transiently expresses an Oct4 polypeptide.
13. The method of claim 4 , wherein the mammalian fibroblast can express an Oct4 polypeptide from a replication-defective expression vector.
14. The method of claim 4 , further comprising administering one or more of the cells produced according to the method of claim 4 to a subject.
15. The method of claim 14 , furthering comprising administering at least about 100 of the cells produced according to the method of claim 4 to a subject.
16. The method of claim 14 , wherein the subject suffers or is suspected of suffering from a heart condition or disease.
17. The method of claim 14 , wherein the subject's heart is abnormally enlarged, thickened and/or stiffened.
18. The method of claim 14 , wherein the subject suffers from, or is suspected of suffering from, congestive heart failure.
19. The method of claim 14 , wherein the subject suffers from, or is suspected of suffering from, myocardial infarction.
20. The method of claim 1 , wherein the fibroblast is contacted with a vector for expression of Oct4 polypeptides.
21. The method of claim 1 , wherein the fibroblast is contacted with Oct4 RNA molecules.
22. The method of claim 1 , wherein the fibroblast comprises an introduced RNA comprising an open reading frame (ORF) for the Oct4 polypeptide flanked by a 5′ untranslated region (UTR) containing a translational initiation signal.
23. The method of claim 20 , wherein the fibroblast expresses an Oct4 polypeptide from a heterologous promoter.
24. The method of claim 20 , wherein the fibroblast transiently expresses an Oct4 polypeptide.
25. The method of claim 20 , wherein a heterologous Oct4 nucleic acid is not integrated into the fibroblast's genome.
26. The method of claim 20 , wherein the fibroblast can express an Oct4 polypeptide from a replication-defective expression vector.
27. The method of claim 1 , furthering comprising administering one or more of the cells produced according to the method of claim 1 to a subject.
28. The method of claim 27 , furthering comprising administering at least about 100 of the cells produced according to the method of claim 1 to a subject.
29. The method of claim 28 , wherein the subject suffers or is suspected of suffering from a heart condition or disease.
30. The method of claim 28 , wherein the subject's heart is abnormally enlarged, thickened and/or stiffened.
31. The method of claim 28 , wherein the subject suffers from, or is suspected of suffering from, congestive heart failure.
32. The method of claim 28 , wherein the subject suffers from, or is suspected of suffering from, myocardial infarction.