Folate Receptor Alpha Upregulates Oct4, Sox2 and Klf4 and Downregulates miR-138 and miR-let-7 in Cranial Neural Crest Cells.
Prenatal folic acid (FA) supplementation prevents neural tube defects. Folate receptor alpha (FRα) is critical for embryonic development, including neural crest (NC) development. Previously we showed that FRα translocates to the nucleus in response to FA, where it acts as a transcription factor. In this study, we examined if FA through interaction with FRα regulates stem cell characteristics of cranial neural crest cells (CNCCs)-critical for normal development. We hypothesized that FRα upregulates coding genes and simultaneously downregulates non-coding miRNA which targets coding genes in CNCCs. Quantitative RT-PCR and chromatin immunoprecipitation showed that FRα upregulates Oct4, Sox2, and Klf4 by binding to their cis-regulator elements-5' enhancer/promoters defined by H3K27Ac and p300 occupancy. FA via FRα downregulates miRNAs, miR-138 and miR-let-7, which target Oct4 and Trim71 (an Oct4 downstream effector), respectively. Co-immunoprecipitation data suggests that FRα interacts with the Drosha-DGCR8 complex to affect pre-miRNA processing. Transfecting anti-miR-138 or anti-miR-let-7 into non-proliferating neural crest cells (NCCs) derived from Splotch (Sp-/-), restored their proliferation potential. In summary, these results suggest a novel pleiotropic role of FRα: (a) direct activation of Oct4, Sox2, and Klf4 genes; and (b) repression of biogenesis of miRNAs that target these genes or their effector molecules. Stem Cells 2016;34:2721-2732.
Developmental Biology Program, Stanley Manne Children's Research Institute, Division of Pediatric Neurosurgery, Ann and Robert H. Lurie Children's Hospital of Chicago, Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
Vineet
V
Mohanty
Developmental Biology Program, Stanley Manne Children's Research Institute, Division of Pediatric Neurosurgery, Ann and Robert H. Lurie Children's Hospital of Chicago, Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
Amar
A
Shah
Developmental Biology Program, Stanley Manne Children's Research Institute, Division of Pediatric Neurosurgery, Ann and Robert H. Lurie Children's Hospital of Chicago, Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
Elise
E
Allender
Developmental Biology Program, Stanley Manne Children's Research Institute, Division of Pediatric Neurosurgery, Ann and Robert H. Lurie Children's Hospital of Chicago, Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
M Rizwan
MR
Siddiqui
Developmental Biology Program, Stanley Manne Children's Research Institute, Division of Pediatric Neurosurgery, Ann and Robert H. Lurie Children's Hospital of Chicago, Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
Sarah
S
Monick
Department of Neurosurgery, Japanese Red Cross Medical Center, Shibuya-Ku, Tokyo, Japan.
Shunsuke
S
Ichi
Department of Biology, Purdue University at Calumet, Hammond, Indiana, USA.
Barbara
B
Mania-Farnell
Developmental Biology Program, Stanley Manne Children's Research Institute, Division of Pediatric Neurosurgery, Ann and Robert H. Lurie Children's Hospital of Chicago, Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
David
D
G McLone
Developmental Biology Program, Stanley Manne Children's Research Institute, Division of Pediatric Neurosurgery, Ann and Robert H. Lurie Children's Hospital of Chicago, Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
Tadanori
T
Tomita
Developmental Biology Program, Stanley Manne Children's Research Institute, Division of Pediatric Neurosurgery, Ann and Robert H. Lurie Children's Hospital of Chicago, Department of Neurological Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
Chandra Shekhar
CS
Mayanil
D000818:Animals; D000070416:Antagomirs / Q000235:genetics / Q000378:metabolism; D005260:Female; D058975:Folate Receptor 1 / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D005492:Folic Acid / Q000378:metabolism / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental; D006657:Histones / Q000235:genetics / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D009432:Neural Crest / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D050814:Octamer Transcription Factor-3 / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D000072041:PAX3 Transcription Factor / Q000172:deficiency / Q000235:genetics; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D043244:Ribonuclease III / Q000235:genetics / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D050880:p300-CBP Transcription Factors / Q000235:genetics / Q000378:metabolism; D000099224:p300-CBP-Associated Factor
D000070416:Antagomirs; D058975:Folate Receptor 1; D005492:Folic Acid; D006657:Histones; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D035683:MicroRNAs; D050814:Octamer Transcription Factor-3; D000072041:PAX3 Transcription Factor; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D043244:Ribonuclease III; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C474044:Dgcr8 protein, mouse; C520574:Folr1 protein, mouse; C000715468:Klf4 protein, mouse; C511366:LIN-41 protein, mouse; C583174:MIRN138 microRNA, mouse; C494174:Pou5f1 protein, mouse; C527783:Sox2 protein, mouse; C542687:mirnlet7 microRNA, mouse; C068686:Pax3 protein, mouse; C513400:Drosha protein, mouse
Cranial neural crest cells; Enhancer/promoters; Folate receptor alpha; Klf4; Multipotency; Oct4; Sox2; miR-138; miR-let-7