Induction of Human Trophoblast Stem Cells from Somatic Cells and Pluripotent Stem Cells

Cell Reports, Journal Year: 2020, Volume and Issue: 33(8), P. 108419 - 108419

Published: Nov. 1, 2020

Human trophoblast stem cells (hTSCs) derived from blastocysts and first-trimester cytotrophoblasts offer an unprecedented opportunity to study the placenta. However, access human embryos placentas is limited, thus preventing establishment of hTSCs diverse genetic backgrounds associated with placental disorders. Here, we show that can be generated numerous using post-natal via two alternative methods: (1) somatic cell reprogramming adult fibroblasts OCT4, SOX2, KLF4, MYC (OSKM) (2) fate conversion naive extended pluripotent cells. The resulting induced/converted recapitulated hallmarks including long-term self-renewal, expression specific transcription factors, transcriptomic signature, potential differentiate into syncytiotrophoblast extravillous We also clarified developmental stage these resemble day 8 cytotrophoblasts. Altogether, hTSC lines origins open possibility model both development diseases in a dish.

Language: Английский

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Therapeutic targeting of TEAD transcription factors in cancer

Trends in Biochemical Sciences, Journal Year: 2023, Volume and Issue: 48(5), P. 450 - 462

Published: Jan. 26, 2023

Language: Английский

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Control of stem cell renewal and fate by YAP and TAZ

Nature Reviews Molecular Cell Biology, Journal Year: 2023, Volume and Issue: 24(12), P. 895 - 911

Published: Aug. 25, 2023

Language: Английский

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Single-cell assessment of primary and stem cell-derived human trophoblast organoids as placenta-modeling platforms

Developmental Cell, Journal Year: 2024, Volume and Issue: 59(6), P. 776 - 792.e11

Published: Feb. 14, 2024

Language: Английский

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TEAD family transcription factors in development and disease

Development, Journal Year: 2021, Volume and Issue: 148(12)

Published: June 15, 2021

ABSTRACT The balance between stem cell potency and lineage specification entails the integration of both extrinsic intrinsic cues, which ultimately influence gene expression through activity transcription factors. One example this is provided by Hippo signalling pathway, plays a central role in regulating organ size during development. pathway mediated transcriptional co-factors Yes-associated protein (YAP) co-activator with PDZ-binding motif (TAZ), interact TEA domain (TEAD) proteins to regulate expression. Although roles YAP TAZ have been intensively studied, played TEAD are less well understood. Recent studies begun address this, revealing that TEADs progenitor self-renewal differentiation throughout various stages Furthermore, it becoming apparent other biology. This Primer provides an overview development, focusing on their as within developmental, homeostatic disease contexts.

Language: Английский

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Characterization of primary models of human trophoblast

Development, Journal Year: 2021, Volume and Issue: 148(21)

Published: Oct. 15, 2021

ABSTRACT Two recently developed models, trophoblast organoids and stem cells (TSCs), are useful tools to further the understanding of human placental development. Both differentiate from villous cytotrophoblast (VCT) either extravillous (EVT) or syncytiotrophoblast (SCT). Here, we compare transcriptomes miRNA profiles these models identify which they resemble in vivo. Our findings indicate that TSCs do not readily undergo SCT differentiation closely at base cell columns where EVT derives. In contrast, similar VCT spontaneous differentiation. A defining feature is leukocyte antigen (HLA) null, whereas expresses HLA-C, -G -E molecules. We find retain vivo characteristics. express classical HLA-A HLA-B molecules, maintain their expression after differentiation, with upregulation HLA-G. Furthermore, HLA differs when grown 3D rather than 2D, suggesting mechanical cues important. results can be used select most suitable model for study development, function pathology.

Language: Английский

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Two distinct trophectoderm lineage stem cells from human pluripotent stem cells

Journal of Biological Chemistry, Journal Year: 2021, Volume and Issue: 296, P. 100386 - 100386

Published: Jan. 1, 2021

The trophectoderm layer of the blastocyst-stage embryo is precursor for all trophoblast cells in placenta. Human stem (TS) have emerged as an attractive tool studies on early development. However, use TS cell models constrained by limited genetic diversity existing lines and restrictions using human fetal tissue or embryos needed to generate additional lines. Here we report derivation two distinct types lineage from pluripotent cells. Analogous villous cytotrophoblasts vivo, first a CDX2- comparable with placenta-derived cells—they both exhibit identical expression key markers, are maintained culture differentiate under similar conditions, share high transcriptome similarity. second CDX2+ requirements, differences gene differentiation, relative Derivation will significantly enable construction vitro normal pathological placental Specification inner mass differentiation event during embryonic mediates blastocyst implantation uterus Upon implantation, forms cytotrophoblast (CTB), putative that can form major placenta, extravillous (EVT) syncytiotrophoblast (STB) (1Bischof P. Irminger-Finger I. cytotrophoblastic cell, mononuclear chameleon.Int. J. Biochem. Cel. Biol. 2005; 37: 1-16Crossref PubMed Scopus (125) Google Scholar, 2Benirschke Kurt. Baergen R.N. Burton G. Graham Pathology Placenta [electronic Resource]. Springer, Heidelberg2012Crossref (48) Scholar). EVTs involved remodeling uterine arteries, which critical ensure adequate perfusion placenta maternal blood, whereas multinucleated STB nutrient gas exchange at maternal–fetal interface (3Yabe S. Alexenko A.P. Amita M. Yang Y. Schust D.J. Sadovsky Ezashi T. Roberts R.M. Comparison generated term placentas.Proc. Natl. Acad. Sci. U. A. 2016; 113: E2598-E2607Crossref (68) 4Moser Orendi K. Gauster Siwetz Helige C. Huppertz B. art identification trophoblast.Placenta. 2011; 32: 197-199Crossref (29) Abnormalities development associated pregnancy-related pathologies such miscarriage, preeclampsia, accreta. Yet, despite its relevance health, constraints research impede mechanistic insight into Trophoblast derived first-trimester samples model system (5Okae H. Toh Sato Hiura Takahashi Shirane Kabayama Suyama Sasaki Arima cells.Cell cell. 2018; 22: 50-63.e6Abstract Full Text PDF (233) restricted accessibility gestation low limit this model. In contrast, (hPSCs) more accessible source generating trophoblast. Of importance, unlike primary where projected pregnancy outcome uncertain, induced (hiPSCs) potentially provide validated (6Sheridan M.A. Jain Lyons A.S. Brahmasani S.R. Dai Tian Ellersieck M.R. Tuteja Schulz L.C. Early onset preeclampsia trophoblast.Proc. 2019; 116: 4336-4345Crossref (20) whether bona fide be obtained hPSCs has been subject intense debate (7Roberts Loh K.M. Bernardo Adachi Telugu B.P.V.L. Pedersen R.A. Differentiation cells: To not be?.Reproduction (Cambridge, England). 2014; 147: D1-D12Crossref (45) A rigorous head-to-head comparison between their vivo counterparts proven difficult owing multiple reasons. Previous used varying experimental protocols (8Roberts Sheridan exposed BMP4†.Biol. Reprod. 99: 212-224Crossref (22) Scholar); cultures terminally differentiated heterogeneity contain many types, until recently self-renewing TS-like had (9Dong Beltcheva Gontarz Zhang Popli Fischer L.A. Khan S.A. Park K.-M. Yoon E.-J. Xing X. Kommagani R. Wang Solnica-Krezel L. Theunissen T.W. naïve cells.eLife. 2020; 9: e52504Crossref (57) 10Cinkornpumin J.K. Kwon S.Y. Guo Hossain Sirois Russett C.S. Tseng H.W. Okae Duchaine T.F. Liu W. Pastor W.A. Naive give rise trophoblast-like Methylome.Stem Cell Rep. 15: 198-213Abstract (33) 11Li Z. Kurosawa O. Iwata Development cystic structures limited-area culture.Biochem. Biophysical Res. Commun. 505: 671-676Crossref (5) 12Gao Nowak-Imialek Chen D. Herrmann Ruan A.C.H. Eckersley-Maslin Ahmad Lee Y.L. Kobayashi Ryan Zhong Zhu Wu et al.Establishment porcine expanded potential cells.Nat. 21: 687-699Crossref (120) study, maintenance hPSCs, specifically (hESCs) hiPSCs, chemically defined conditions. early-gestation CTB. type Critically, isolation populations allowed direct hPSCs; genome-wide transcriptomic analysis functional assays demonstrate very similarity placenta- hPSC-derived routine powerful tools Media formulations previous hESCs included components knockout serum replacement (KSR) bovine albumin (BSA) act carriers lipids. Albumin-associated lipids implicated activation G-protein–coupled receptor–mediated signaling (13Yu F.-X. Zhao Panupinthu N. Jewell J.L. Lian L.H. Yuan Tumaneng Li Fu X.-D. Mills G.B. Guan K.-L. Regulation Hippo-YAP pathway G-protein-coupled receptor signaling.Cell. 2012; 150: 780-791Abstract (974) 14Mendelson Evans Hla Sphingosine 1-phosphate signalling.Development 141: 5-9Crossref (165) For instance, phospholipid sphingosine-1 phosphate (S1P) present KSR activate YAP signaling. plays role specification mouse (15Yagi Kohn M.J. Karavanova Kaneko K.J. Vullhorst DePamphilis M.L. Buonanno Transcription factor TEAD4 specifies beginning mammalian development.Development 2007; 134: 3827-3836Crossref (353) 16Knott J.G. Paul Transcriptional regulators mammals hemochorial placentation.Reproduction 148: R121-R136Crossref (40) 17Nishioka Yamamoto Kiyonari Sawada Ota Nakao Tead4 required pre-implantation embryos.Mech. Dev. 2008; 125: 270-283Crossref (331) Scholar), well (18Saha Ganguly Home Bhattacharya Ray Ghosh Rumi M.A.K. Marsh French V. Gunewardena ensures postimplantation promoting self-renewal: An implication loss.Proc. 117: 202002449Crossref 19Meinhardt Haider Kunihs Saleh Pollheimer Fiala Hetey Feher Szilagyi Than N.G. Knöfler Pivotal transcriptional co-activator stemness developing placenta.Proc. 13562-13570Crossref (25) We investigated S1P context modifying our protocol utilized (20Sarkar Randall S.M. Collier T.S. Nero Russell T.A. Muddiman D.C. Rao B.M. Activin/nodal Switches terminal fate cell-derived trophoblasts.J. Chem. 2015; 290: 8834-8848Abstract (15) 21Sarkar Mischler Dorman K.F. Boggess K.A. Identification epigenetic proteins expressed trophoblasts Proteome 2433-2444Crossref (6) H1 H9 cultured E8 medium were 6 days E7 (E8 without transforming growth factor-beta1 [TGFβ1]) supplemented S1P, treatment BMP4 activin/nodal inhibitor SB431542 (Fig. 1A). Under these observed upregulation marker CDX2 CTB ELF5 S1, B). Upregulation TBX4 was after days. overall there no significant changes markers neural mesodermal suggesting lineages did occur Immunofluorescence day confirmed pan-trophoblast KRT7, P63 GATA3; (Figs. 1B S1C). ability STB, those previously employed formation mesenchymal epithelial over 6-day period when passaged epidermal (EGF) SB431542. showed KRT7 EVT VE-Cadherin HLA-G 1C, S1D). Alternatively, passaging CTB-like E6 TGFβ1 fibroblast factor-2 [FGF2]) activin EGF resulted KRT7+ multinucleate expressing hCG syncytin 8-day 1D, S1E). Removal hESC abolished express 1E, S2A) conditions also absence evidenced lack 1F, S2B). Also, downregulation transcripts mesoderm upon removal S2C). Taken together results show cells, utilizing complex containing S1P. Furthermore, addition exogenous necessary medium. Rho GTPase signaling, downstream receptors activated nuclear localization (22Ohgushi Minaguchi Sasai Rho-signaling-directed YAP/TAZ activity Underlies long-term Survival Expansion 17: 448-461Abstract (100) 23Mo J.-S. Yu Gong Brown J.H. protease-activated (PARs).Genes 26: 2138-2143Crossref (195) Both Rho/RhoA kinase (ROCK) play (24Nishioka Inoue Ralston Yabuta Hirahara Stephenson R.O. Ogonuki Makita Kurihara Morin-Kensicki E.M. Nojima Rossant al.The Hippo Lats Yap pattern distinguish mass.Dev. 2009; 16: 398-410Abstract (657) 25Kono Tamashiro D.A.A. Alarcon V.B. Inhibition RHO-ROCK enhances ICM suppresses TE characteristics through blastocyst.Dev. 394: 142-155Crossref (79) Therefore, Rho/ROCK hESCs. Y-27632 subsequent investigate hESCs; however, weak only few S3A). On other hand, addition, presence ROCK inhibition formation, shown S3B). hESCs, line (H9) expresses inducible shRNA against (H9-YAP-ishRNA) scrambled control (26Hsiao Lampe Nillasithanukroh Han Palecek S.P. density modulates signaling.Biotechnol. 11: 662-675Crossref (26) knockdown relevant markers. It notable death S3, Gene revealed reduction knockdown, S3C). Significant genes LMO2 observed, T upregulated, H9-YAP-ishRNA, control. together, acts receptor-mediated receptor-independent pathways (14Mendelson 27Maceyka Harikumar K.B. Milstien Spiegel Sphingosine-1-phosphate disease.Trends 50-60Abstract (695) specific mechanism action trophoblast, replaced D-erythro-dihydrospingosine-1-phosphate (dhS1P) protocol. dhS1P agonist (S1PRs) but does mediate intracellular effect (28Van Brocklyn J.R. M.-J. Menzeleev Olivera Edsall Cuvillier Thomas D.M. Coopman P.J.P. Thangada C.H. Dual actions sphingosine-1-phosphate: Extracellular G i -coupled Edg-1 regulate proliferation Survival.J. 1998; 142: 229-240Crossref (444) Replacing yielded results—CTB-like CDX2, GATA3, P63, 2A S4A). further described 1A), hCG, 2, B C; Fig. S4, C). These suggest S1PR extracellularly S1PR1-5 TBs S1PR1-3 (29Johnstone E.D. Chan Sibley C.P. Davidge S.T. Lowen Guilbert L.J. G(i)-coupled response.J. lipid 46: 1833-1839Abstract (34) selective chemical agonists S1PR1-3—CYM5442 hydrochloride, CYM5520, CYM5541, respectively—to replace discussed. Expression three Similarly, each some variability S5). S1PR2 strong cytoplasmic staining agonists. Formation large pronounced S1PR3 used, compared S1PR1 agonist. enhanced EVTs, effects system. Since qualitative observations could chose studies. CYM5541 blastocyst- plating (TSCM) developed al. hESC-derived underwent colonies retained single passage. upregulated little 2 initiation decreases reported CDX2+/p63+ BMP 4 (30Horii Wakeland A.K. Pizzo D.P. Nelson K.K. Sabatini Laurent Parast M.M. disease.Proc. United States America. E3882-E3891Crossref (66) explored shorter step obtaining 3A). After 3 uniformly 3B). most lose 3C). Quantitative image nearly 3, contrast 6. note, reduced fraction case 3-day protocol; 3D). Transcriptome RNA sequencing identified 291 higher levels 330 lower versus undifferentiated (Tables S1 S2).Expression trophectoderm-associated HAND1, TFAP2A, pluripotency-associated NANOG downregulated. set enrichment differentially 567 202 ontology (GO) categories (of 9996 queried categories) respectively S3 S4). Consistent GO terms epithelium development, proliferation, four (denoted TM4), GSK3β CHIR99021, TGFβ A83-01, FGF10. CHIR99021 A83-01 TSCM cells; FGF10 because FGFR2b active Cells TM4 30+ passages course 5 months. medium, TFAP2C, YAP, TEAD4, GATA3 3E S6) 31Choi Carey Wilson C.A. Knott AP-2γ core regulator tight junction biogenesis cavity embryogenesis.Development 139: 4623-4632Crossref 32Home Saha Dutta Yoo Pal Vivian Larson Petroff Gallagher P.G. V.P. White K.L. Golos T.G. Behr al.Altered subcellular transcription regulates commitment.Proc. 109: 7362-7367Crossref (113) 33Ralston Cox B.J. Nishioka Chea E. Rugg-Gunn Robson Draper J.S. Gata3 parallel Cdx2.Development 2010; 137: 395-403Crossref (299) 34Niwa Toyooka Shimosato Strumpf Yagi Interaction Oct3/4 Cdx2 determines differentiation.Cell. 123: 917-929Abstract (854) P63. strongly lost once villi formed 35Hemberger Udayashankar Tesar Moore G.J. ELF5-enforced networks define epigenetically regulated compartment placenta.Hum. Mol. Genet. 19: 2456-2467Crossref (126) 36Blakeley Fogarty N.M.E. del Valle Wamaitha S.E. Hu T.X. Elder Snell Christie Niakan Defining single-cell RNA-seq.Development. 3151-3165Crossref (262) 37Knöfler Gamage T.K.J.B. James development: Key molecular mechanisms systems.Cell Life 76: 3479-3496Crossref (144) indicate do denoted hPSC-TSCDX2 evaluated same able 3F). treatment, elongated acquired flattened morphology. passage, minimal 3G). morp

Language: Английский

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How trophoblasts fuse: an in-depth look into placental syncytiotrophoblast formation

Cellular and Molecular Life Sciences, Journal Year: 2022, Volume and Issue: 79(8)

Published: July 20, 2022

Language: Английский

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Cell trajectory modeling identifies a primitive trophoblast state defined by BCAM enrichment

Development, Journal Year: 2022, Volume and Issue: 149(1)

Published: Jan. 1, 2022

ABSTRACT In early placental development, progenitor cytotrophoblasts (CTB) differentiate along one of two cellular trajectories: the villous or extravillous pathways. CTB committed to pathway fuse with neighboring form outer multinucleated syncytiotrophoblast (SCT), whereas into invasive trophoblasts (EVT). Unfortunately, little is known about processes controlling human maintenance and differentiation. To address this, we established a single cell RNA sequencing (scRNA-seq) dataset from first trimester placentas identify states important in trophoblast establishment, renewal Multiple distinct were identified, representing CTB, column SCT precursors EVT. Lineage trajectory analysis identified origin that was reproduced stem organoids. Heightened expression basal adhesion molecule (BCAM) defined this primitive state, where BCAM enrichment gene silencing resulted enhanced diminished organoid growth, respectively. Together, work describes at high-resolution heterogeneity within trimester, resolves networks progenitors identifies as marker possible regulator.

Language: Английский

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Derivation of functional trophoblast stem cells from primed human pluripotent stem cells

Stem Cell Reports, Journal Year: 2022, Volume and Issue: 17(6), P. 1303 - 1317

Published: May 19, 2022

Trophoblast stem cells (TSCs) have recently been derived from human embryos and early-first-trimester placenta; however, aside ethical challenges, the unknown disease potential of these limits their scientific utility. We previously established a bone morphogetic protein 4 (BMP4)-based two-step protocol for differentiation primed pluripotent (hPSCs) into functional trophoblasts; those trophoblasts could not be maintained in self-renewing TSC-like state. Here, we use first step this protocol, followed by switch to newly developed TSC medium, derive bona fide TSCs. show that resemble placenta- naive hPSC-derived TSCs, based on transcriptome as well vitro vivo potential. conclude hPSCs can used generate TSCs through simple which applied widely available set existing hPSCs, including induced cells, patients with known birth outcomes.

Language: Английский

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