Enhancers of induced pluripotent stem cell reprogramming
Described herein, inter alia, are methods and compositions useful for induced pluripotent stem cell reprogramming.
1. An in vitro method of making an induced pluripotent stem cell comprising:
(i) contacting a non-pluripotent cell with a compound having the formula:
wherein,
X 1 is C(R 12 ) or N;
X 2 is C(R 4 ) or N;
X 3 is C(R 5 ) or N;
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl, wherein R 2 and R 3 are optionally joined to form a substituted or unsubstituted heterocycloalkyl or substituted or unsubstituted heteroaryl; and
(ii) transfecting said non-pluripotent cell with a nucleic acid encoding a Klf4 protein, introducing a recombinant Klf4 protein into said non-pluripotent cell, or contacting said non-pluripotent cell with a Klf4 expression inducer;
(iii) transfecting said non-pluripotent cell with a nucleic acid encoding a Sox2 protein, introducing a recombinant Sox2 protein into said non-pluripotent cell, or contacting said non-pluripotent cell with a Sox2 expression inducer;
(iv) transfecting said non-pluripotent cell with a nucleic acid encoding a cMyc protein, introducing a recombinant cMyc protein into said non-pluripotent cell, or contacting said non-pluripotent cell with a cMyc expression inducer; and
(v) after steps (i), (ii), (iii), and (iv), allowing said non-pluripotent cell to divide thereby forming said induced pluripotent stem cell.
2. The method of claim 1 , further comprising, prior to step (v), transfecting said non-pluripotent cell with a nucleic acid encoding an Oct4 protein or introducing a recombinant Oct4 protein into said non-pluripotent cell.
3. The method of claim 1 , further comprising, prior to step (v), transfecting said non-pluripotent cell with a nucleic acid encoding a Nanog protein or introducing a recombinant Nanog protein into said non-pluripotent cell.
4. The method of claim 1 , further comprising, prior to step (v), transfecting said non-pluripotent cell with a nucleic acid encoding a Lin28 protein, introducing a recombinant Lin28 protein into said non-pluripotent cell, or contacting said non-pluripotent cell with a Lin28 expression inducer.
5. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, or substituted or unsubstituted heterocycloalkyl.
6. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , substituted or unsubstituted alkyl, or substituted or unsubstituted heteroalkyl.
7. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , substituted or unsubstituted C 1 to C 10 alkyl, substituted or unsubstituted 2 to 10 membered heteroalkyl or substituted or unsubstituted 3 to 8membered heterocycloalkyl.
8. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , substituted or unsubstituted C 1 to C 10 alkyl or substituted or unsubstituted 2 to 10 membered heteroalkyl.
9. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , unsubstituted alkyl, unsubstituted heteroalkyl, or substituted heterocycloalkyl.
10. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , unsubstituted alkyl or unsubstituted heteroalkyl.
11. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , unsubstituted C 1 to C 10 alkyl, unsubstituted 2 to 10 membered heteroalkyl, or substituted 3 to 8 membered heterocycloalkyl.
12. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9, R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —CN, —NO 2 , —NH 2 , —CF 3 , —CCl 3 , —OH, —SH, —SO 3 H, —C(O)OH, —C(O)NH 2 , unsubstituted C 1 to C 10 alkyl or unsubstituted 2 to 10 membered heteroalkyl.
13. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, unsubstituted C 1 to C 10 alkyl or unsubstituted 2 to 10 membered heteroalkyl.
14. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —N(CH 3 ) 2 , unsubstituted C 1 to C 5 alkyl or unsubstituted C 1 to C 5 alkoxy.
15. The method of claim 1 , wherein
R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 , R 10 , R 11 and R 12 are independently selected from hydrogen, halogen, —N(CH 3 ) 2 , unsubstituted C 1 to C 5 alkyl, methoxy, ethoxy or propoxy.
16. The method of claim 1 , wherein the compound is selected from the group consisting of
17. The method of claim 1 , wherein the compound is selected from the group consisting of
18. The method of claim 1 , wherein the compound is selected from the group consisting of
19. The method of claim 1 , wherein the compound is selected from the group consisting of
20. The method of claim 1 , wherein R 2 and R 3 are joined together to form a substituted or unsubstituted heteroaryl.
21. The method of claim 2 , wherein R 2 and R 3 are joined together to form a substituted or unsubstituted five-membered heteroaryl.
22. The method of claim 1 , wherein step (ii) is contacting said non-pluripotent cell with a Klf4 expression inducer.
23. The method of claim 22 , wherein said Klf4 expression inducer is valproic acid, kenpaullone, 5-aminoimidazole-4-carboxamide-I-b-riboside, AMI-5, sodium butyrate, PS48, A-83-1, PD0325901, or SB431542.
24. The method of claim 1 , wherein step (iii) is contacting said non-pluripotent cell with a Sox2 expression inducer.
25. The method of claim 24 , wherein said Sox2 expression inducer is BIX, BayK, CHIR99021, tranylcypromine, E-616452, Dasatinib, iPY, PP1, n-Butylidenephthalide, AMI-5, PS48, sodium butyrate, A-83-1, PD0325901, SB431542, an Alk5 inhibitor, and LIF.
26. The method of claim 1 , wherein step (iv) is contacting said non-pluripotent cell with a cMyc expression inducer.
27. The method of claim 26 , wherein said cMyc expression inducer is CHIR99021, BIX, BayK, valproic acid, 5′-azacytidine, 5-aminoimidazole-4-carboxamide-I-b-riboside, AMI-5, sodium butyrate, PS48, A-83-1, SB431542, PD0325901, an Alk5 inhibitor, and LIF.
28. The method of claim 1 , further comprising contacting said non-pluripotent cell with a Lin28 expression inducer.
29. The method of claim 27 , wherein said Lin28 expression inducer is CHIR99021, BIX, BayK, E-616452, Dasatinib, iPY, PP1, valproic acid, 5′-azacytidine, AMI-5, A-83-01, sodium butyrate, PS48, PD0325901, or SB431542.