Human Amniocytes Are Receptive to Chemically Induced Reprogramming to Pluripotency.
Restoring pluripotency using chemical compounds alone would be a major step forward in developing clinical-grade pluripotent stem cells, but this has not yet been reported in human cells. We previously demonstrated that VPA_AFS cells, human amniocytes cultivated with valproic acid (VPA) acquired functional pluripotency while remaining distinct from human embryonic stem cells (hESCs), questioning the relationship between the modulation of cell fate and molecular regulation of the pluripotency network. Here, we used single-cell analysis and functional assays to reveal that VPA treatment resulted in a homogeneous population of self-renewing non-transformed cells that fulfill the hallmarks of pluripotency, i.e., a short G1 phase, a dependence on glycolytic metabolism, expression of epigenetic modifications on histones 3 and 4, and reactivation of endogenous OCT4 and downstream targets at a lower level than that observed in hESCs. Mechanistic insights into the process of VPA-induced reprogramming revealed that it was dependent on OCT4 promoter activation, which was achieved independently of the PI3K (phosphatidylinositol 3-kinase)/AKT/mTOR (mammalian target of rapamycin) pathway or GSK3β inhibition but was concomitant with the presence of acetylated histones H3K9 and H3K56, which promote pluripotency. Our data identify, for the first time, the pluripotent transcriptional and molecular signature and metabolic status of human chemically induced pluripotent stem cells.
Institute for Women's Health, Maternal and Fetal Medicine Department, University College London (UCL), London WC1E 6HX, UK.
Kate E
KE
Hawkins
Institute for Women's Health, Maternal and Fetal Medicine Department, University College London (UCL), London WC1E 6HX, UK.
Dafni
D
Moschidou
Department of Comparative Biomedical Sciences, The Royal Veterinary College (RVC), Royal College Street, London NW1 0TU, UK.
Danilo
D
Faccenda
Institute for Stem Cell Research and Regenerative Medicine, Heinrich Heine University Dusseldorf, Dusseldorf 40225, Germany.
Wasco
W
Wruck
Imprinting and Cancer Group, Cancer Epigenetic and Biology Program, Bellvitge Institute for Biomedical Research (IDIBELL), Hospital Duran i Reynals, Barcelona 08908, Spain.
Alex
A
Martin-Trujillo
Institute for Women's Health, Maternal and Fetal Medicine Department, University College London (UCL), London WC1E 6HX, UK; Imperial College London, National Heart and Lung Institute, Hammersmith Campus, Du Cane Road, London W12 0NN, UK.
Kwan-Leong
KL
Hau
Institute for Women's Health, Maternal and Fetal Medicine Department, University College London (UCL), London WC1E 6HX, UK.
Anna Maria
AM
Ranzoni
Departments of Medicine and Radiology, Stanford University, Stanford, CA 94305-5454, USA.
Veronica
V
Sanchez-Freire
Institute for Women's Health, Maternal and Fetal Medicine Department, University College London (UCL), London WC1E 6HX, UK; Institute for Child Health, University College London, London WC1N 1EH, UK.
Fabio
F
Tommasini
Institute for Child Health, University College London, London WC1N 1EH, UK.
Simon
S
Eaton
Institute for Child Health, University College London, London WC1N 1EH, UK.
Paolo
P
De Coppi
Institute for Stem Cell Research and Regenerative Medicine, Heinrich Heine University Dusseldorf, Dusseldorf 40225, Germany.
David
D
Monk
Department of Comparative Biomedical Sciences, The Royal Veterinary College (RVC), Royal College Street, London NW1 0TU, UK; Consortium for Mitochondrial Research, University College London, Royal College Street, London NW1 0TU, UK.
Michelangelo
M
Campanella
Institute for Child Health, University College London, London WC1N 1EH, UK.
Adrian J
AJ
Thrasher
Institute for Stem Cell Research and Regenerative Medicine, Heinrich Heine University Dusseldorf, Dusseldorf 40225, Germany.
James
J
Adjaye
Institute for Women's Health, Maternal and Fetal Medicine Department, University College London (UCL), London WC1E 6HX, UK. Electronic address: p.guillot@ucl.ac.uk.
Pascale V
PV
Guillot
D000650:Amnion / Q000166:cytology*; D015415:Biomarkers; D002453:Cell Cycle / Q000235:genetics; D054338:Cell Transdifferentiation / Q000187:drug effects* / Q000235:genetics; D065150:Cellular Reprogramming / Q000187:drug effects* / Q000235:genetics; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D004734:Energy Metabolism; D044127:Epigenesis, Genetic; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D006019:Glycolysis; D006657:Histones / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D000071317:Nanog Homeobox Protein / Q000235:genetics; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D010641:Phenotype; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D015533:Transcriptional Activation
D015415:Biomarkers; D006657:Histones; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D011993:Recombinant Fusion Proteins; D058570:TOR Serine-Threonine Kinases
OCT4; amniotic stem cells; biotechnology; cell biology; chemical reprogramming; developmental biology; medical research; plasticity; pluripotency; stem cells