Efficient direct reprogramming of mature amniotic cells into endothelial cells by ETS factors and TGFβ suppression.
ETS transcription factors ETV2, FLI1, and ERG1 specify pluripotent stem cells into induced vascular endothelial cells (iVECs). However, iVECs are unstable and drift toward nonvascular cells. We show that human midgestation c-Kit(-) lineage-committed amniotic cells (ACs) can be reprogrammed into vascular endothelial cells (rAC-VECs) without transitioning through a pluripotent state. Transient ETV2 expression in ACs generates immature rAC-VECs, whereas coexpression with FLI1/ERG1 endows rAC-VECs with a vascular repertoire and morphology matching mature endothelial cells (ECs). Brief TGFβ-inhibition functionalizes VEGFR2 signaling, augmenting specification of ACs into rAC-VECs. Genome-wide transcriptional analyses showed that rAC-VECs are similar to adult ECs in which vascular-specific genes are expressed and nonvascular genes are silenced. Functionally, rAC-VECs form stable vasculature in Matrigel plugs and regenerating livers. Therefore, short-term ETV2 expression and TGFβ inhibition with constitutive ERG1/FLI1 coexpression reprogram mature ACs into durable rAC-VECs with clinical-scale expansion potential. Banking of HLA-typed rAC-VECs establishes a vascular inventory for treatment of diverse disorders.
Howard Hughes Medical Institute, Ansary Stem Cell Institute, Department of Genetic Medicine, Weill Cornell Medical College, New York, NY, 10065, USA.
Michael
M
Ginsberg
Daylon
D
James
Bi-Sen
BS
Ding
Daniel
D
Nolan
Fuqiang
F
Geng
Jason M
JM
Butler
William
W
Schachterle
Venkat R
VR
Pulijaal
Susan
S
Mathew
Stephen T
ST
Chasen
Jenny
J
Xiang
Zev
Z
Rosenwaks
Koji
K
Shido
Olivier
O
Elemento
Sina Y
SY
Rabbany
Shahin
S
Rafii
D000653:Amniotic Fluid / Q000166:cytology*; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology*; D006801:Humans; D050783:Proto-Oncogene Proteins c-ets / Q000378:metabolism*; D015682:Retroviridae Proteins, Oncogenic / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D050783:Proto-Oncogene Proteins c-ets; D015682:Retroviridae Proteins, Oncogenic; D016212:Transforming Growth Factor beta; C050379:oncogene proteins v-ets