Chemical Enhancement of In Vitro and In Vivo Direct Cardiac Reprogramming.
BACKGROUND
Reprogramming of cardiac fibroblasts into induced cardiomyocyte-like cells in situ represents a promising strategy for cardiac regeneration. A combination of 3 cardiac transcription factors, Gata4, Mef2c, and Tbx5 (GMT), can convert fibroblasts into induced cardiomyocyte-like cells, albeit with low efficiency in vitro.


METHODS
We screened 5500 compounds in primary cardiac fibroblasts to identify the pathways that can be modulated to enhance cardiomyocyte reprogramming.


RESULTS
We found that a combination of the transforming growth factor-β inhibitor SB431542 and the WNT inhibitor XAV939 increased reprogramming efficiency 8-fold when added to GMT-overexpressing cardiac fibroblasts. The small molecules also enhanced the speed and quality of cell conversion; we observed beating cells as early as 1 week after reprogramming compared with 6 to 8 weeks with GMT alone. In vivo, mice exposed to GMT, SB431542, and XAV939 for 2 weeks after myocardial infarction showed significantly improved reprogramming and cardiac function compared with those exposed to only GMT. Human cardiac reprogramming was similarly enhanced on transforming growth factor-β and WNT inhibition and was achieved most efficiently with GMT plus myocardin.


CONCLUSIONS
Transforming growth factor-β and WNT inhibitors jointly enhance GMT-induced direct cardiac reprogramming from cardiac fibroblasts in vitro and in vivo and provide a more robust platform for cardiac regeneration.
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Tamer M A
TM
Mohamed
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Nicole R
NR
Stone
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Emily C
EC
Berry
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Ethan
E
Radzinsky
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Yu
Y
Huang
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Karishma
K
Pratt
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Yen-Sin
YS
Ang
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Pengzhi
P
Yu
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Haixia
H
Wang
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Shibing
S
Tang
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Sergey
S
Magnitsky
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Sheng
S
Ding
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco.
Kathryn N
KN
Ivey
From Gladstone Institute of Cardiovascular Disease and Roddenberry Center for Stem Cell Biology and Medicine, San Francisco, CA (T.M.A.M., N.R.S., E.C.B., E.R., Y.H., K.P., Y.-S.A., P.Y., H.W., S.T., S.D., K.N.I., D.S.); Institute of Cardiovascular Sciences, University of Manchester, UK (T.M.A.M.); Faculty of Pharmacy, Zagazig University, Egypt (T.M.A.M.); and Departments of Radiology (S.M.), Pharmaceutical Chemistry (S.D.), Pediatrics (K.N.I., D.S.), and Biochemistry and Biophysics (D.S.), University of California San Francisco. deepak.srivastava@gladstone.ucsf.edu.
Deepak
D
Srivastava
D000818:Animals; D001549:Benzamides / Q000494:pharmacology* / Q000627:therapeutic use; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000187:drug effects*; D004149:Dioxoles / Q000494:pharmacology* / Q000627:therapeutic use; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism; D006321:Heart / Q000000981:diagnostic imaging; D006575:Heterocyclic Compounds, 3-Ring / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D064326:MEF2 Transcription Factors / Q000235:genetics / Q000378:metabolism; D008279:Magnetic Resonance Imaging; D051379:Mice; D009203:Myocardial Infarction / Q000188:drug therapy; D009206:Myocardium / Q000473:pathology; D032383:Myocytes, Cardiac / Q000166:cytology / Q000378:metabolism; D020825:T-Box Domain Proteins / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors / Q000378:metabolism; D051153:Wnt Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism
C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D004149:Dioxoles; D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse; D006575:Heterocyclic Compounds, 3-Ring; D064326:MEF2 Transcription Factors; D020825:T-Box Domain Proteins; C103172:T-box transcription factor 5; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; D051153:Wnt Proteins; C544261:XAV939
cell differentiation; heart; regeneration; transcription factors