Pluripotent human stem cells as models for normal and diseased trophoblast

Summary

Principal Investigator: ROBERT MICHAEL ROBERTS
Abstract: DESCRIPTION (provided by applicant): The main goal of this proposal is to generate extravillous trophoblast (EVT) from human embryonic stem cells (hESC) after they have been treated with BMP4 (BMP-hESC) and use them as a model to study EVT emergence, migration, and invasiveness, as well as intrinsic and extrinsic factors that influence these properties. This project has high significance because EVT ultimately become the invasive trophoblast (TR) cells of human placenta that penetrate maternal blood vessels and enhance blood flow. While invasion of the uterine decidua and the proximate inner third of the myometrium by EVT is a characteristic feature of a healthy pregnancy, shallow invasion is associated with placental pathologies, among them pre-eclampsia (PE) and intra-uterine growth retardation (IUGR). PE is a pregnancy-specific syndrome that affects ~ 3 % of pregnancies and is responsible for 15 % of pre-term births and an estimated 50,000 deaths per year worldwide. It is characterized by maternal hypertension, edema, and proteinuria, conditions that in the more serious, early onset disease can manifest as soon as 20 weeks of pregnancy. Aim 1 will establish and characterize a hESC-derived cell culture model for the study of EVT. This aim is based on the hypothesis that the sub-population of HLA-G positive BMP-hESC that migrates through a Matrigel-coated matrix under high oxygen (O2) conditions are EVT homologs. HLA-G positive cells that remain associated with the BMP-hESC colonies under low O2 conditions represent a precursor population of non- invasive EVT whose properties will be distinct from the invasive population captured under high O2 conditions. Aim 2 will examine the ability of the EVT derived from BMP-hESC to invade through cross-linked, relatively stiff matrices, such as Type I collagen, and to interact with microcapillaries cultured within such matrices. Aim 3 will examine a possible model for PE in which up-regulation of stable HIF1 alpha (HIF1A) protein is hypothesized to counteract the effects of high O2 during in vitro development of BMP-hESC and retard development into invasive EVT. If time permits or the HIF1A hypothesis proves incorrect, we would plan to study the effects of knockdown of at least one transcription factor that has been implicated in EVT emergence and differentiation, namely ASCL2. Aim 4 will attempt to re-create EVT from infants born to mothers with PE by generating induced pluripotent stem cells (iPSC) from discarded umbilical cord and converting these pluripotent cells to CT, EVT and other lineages by the BMP4 approach. The properties of these cells can then be compared with EVT generated from cord cells from pregnancies not complicated by PE. The hypothesis underpinning this aim is that some forms of PE are initiated by genetically-based placental pathology, and these abnormalities will be manifested in the phenotype of EVT generated from umbilical cord-derived iPSC. Together, these aims will bring innovative and new approaches to the study of human TR cells and to examining the basis of placental pathologies, especially PE. They will create the first in vitro model that makes possible the study of both the emergence and invasiveness of EVT cells, and the first model to study these early events directly in TR from pregnancies complicated by PE. PUBLIC HEALTH RELEVANCE: This project is designed to establish a new model for studying extravillous trophoblast (EVT), a placental cell type that invades the wall of the womb during the first trimester of pregnancy and whose failure to invade properly can lead to serious consequences for mother and child, including a condition called pre-eclampsia. Instead of isolating the cells from placentae, the goal is to generate EVT from human pluripotent stem cells, thereby allowing us to study factors that control their invasiveness. Also by generating such pluripotent cells from umbilical cords of babies born to mothers that developed pre-eclampsia, we hope to recreate the cell type that caused the disease in the first place.
Funding Period: 2010-12-16 - 2015-11-30
more information: NIH RePORT

Top Publications

  1. ncbi Effect of interferon-tau administration on endometrium of nonpregnant ewes: a comparison with pregnant ewes
    Yizhen Chen
    Department of Animal Sciences, University of Missouri, Columbia, 65211, USA
    Endocrinology 147:2127-37. 2006
  2. pmc Cell cycle synchronization of leukemia inhibitory factor (LIF)-dependent porcine-induced pluripotent stem cells and the generation of cloned embryos
    Ye Yuan
    Division of Animal Sciences and Bond Life Sciences Center University of Missouri Columbia, MO USA
    Cell Cycle 13:1265-76. 2014
  3. ncbi Differentiation of trophoblast cells from human embryonic stem cells: to be or not to be?
    R Michael Roberts
    Division of Animal Sciences
    Reproduction 147:D1-12. 2014
  4. pmc Comparison of extravillous trophoblast cells derived from human embryonic stem cells and from first trimester human placentas
    B P Telugu
    University of Maryland, Animal and Avian Sciences, College Park, MD 20742, USA
    Placenta 34:536-43. 2013
  5. pmc Complete and unidirectional conversion of human embryonic stem cells to trophoblast by BMP4
    Mitsuyoshi Amita
    Division of Animal Sciences, and Bond Life Sciences Center, University of Missouri, Columbia, MO 65211, USA
    Proc Natl Acad Sci U S A 110:E1212-21. 2013
  6. ncbi Induced pluripotent stem cells from pigs and other ungulate species: an alternative to embryonic stem cells?
    T Ezashi
    Division of Animal Sciences and Bond Life Sciences Center, University of Missouri Columbia, Columbia, MO, USA
    Reprod Domest Anim 47:92-7. 2012
  7. pmc Squelching of ETS2 transactivation by POU5F1 silences the human chorionic gonadotropin CGA subunit gene in human choriocarcinoma and embryonic stem cells
    Rangan Gupta
    Department of Veterinary Pathobiology, Bond Life Sciences Center, University of Missouri, Columbia, Missouri 65211, USA
    Mol Endocrinol 26:859-72. 2012
  8. pmc Model systems for studying trophoblast differentiation from human pluripotent stem cells
    Toshihiko Ezashi
    Division of Animal Sciences and Bond Life Sciences Center, University of Missouri Columbia, 65211, USA
    Cell Tissue Res 349:809-24. 2012
  9. pmc Preeclampsia: multiple approaches for a multifactorial disease
    Kathleen A Pennington
    Department of Obstetrics, Gynecology and Women s Health, University of Missouri School of Medicine, 500 North Keene Street, Columbia, MO 65201, USA
    Dis Model Mech 5:9-18. 2012
  10. ncbi The contrasting effects of ad libitum and restricted feeding of a diet very high in saturated fats on sex ratio and metabolic hormones in mice
    Andrei P Alexenko
    Department of Animal Sciences, University of Missouri Columbia, Christopher S Bond Life Sciences Center, Columbia, Missouri 65211 7310, USA
    Biol Reprod 77:599-604. 2007

Research Grants

  1. CARDIOVASCULAR DYNAMICS AND THEIR CONTROL
    John E Hall; Fiscal Year: 2013

Detail Information

Publications13

  1. ncbi Effect of interferon-tau administration on endometrium of nonpregnant ewes: a comparison with pregnant ewes
    Yizhen Chen
    Department of Animal Sciences, University of Missouri, Columbia, 65211, USA
    Endocrinology 147:2127-37. 2006
    ..IFNT treatment mimics pregnancy, but relatively high doses of im-injected IFNT are required to elicit a full endometrial response...
  2. pmc Cell cycle synchronization of leukemia inhibitory factor (LIF)-dependent porcine-induced pluripotent stem cells and the generation of cloned embryos
    Ye Yuan
    Division of Animal Sciences and Bond Life Sciences Center University of Missouri Columbia, MO USA
    Cell Cycle 13:1265-76. 2014
    ..Together, these data provide at least a partial explanation for the almost complete failure to produce cloned piglets from piPSC...
  3. ncbi Differentiation of trophoblast cells from human embryonic stem cells: to be or not to be?
    R Michael Roberts
    Division of Animal Sciences
    Reproduction 147:D1-12. 2014
    ..By analogy to other differentiation systems, our debate is broadly applicable, as it articulates higher and more challenging standards for judging whether a given cell type has been genuinely produced from hPSC differentiation. ..
  4. pmc Comparison of extravillous trophoblast cells derived from human embryonic stem cells and from first trimester human placentas
    B P Telugu
    University of Maryland, Animal and Avian Sciences, College Park, MD 20742, USA
    Placenta 34:536-43. 2013
    ..The goals of the present study were to further characterize the HLA-G(+) cells derived from BMP4-treated hESC, and determine their suitability as a model...
  5. pmc Complete and unidirectional conversion of human embryonic stem cells to trophoblast by BMP4
    Mitsuyoshi Amita
    Division of Animal Sciences, and Bond Life Sciences Center, University of Missouri, Columbia, MO 65211, USA
    Proc Natl Acad Sci U S A 110:E1212-21. 2013
    ..The experiments demonstrate that the in vitro BMP4/hESC model is valid for studying the emergence and differentiation of trophoblasts...
  6. ncbi Induced pluripotent stem cells from pigs and other ungulate species: an alternative to embryonic stem cells?
    T Ezashi
    Division of Animal Sciences and Bond Life Sciences Center, University of Missouri Columbia, Columbia, MO, USA
    Reprod Domest Anim 47:92-7. 2012
    ..However, hurdles still remain if such cells are to achieve their biotechnological promise...
  7. pmc Squelching of ETS2 transactivation by POU5F1 silences the human chorionic gonadotropin CGA subunit gene in human choriocarcinoma and embryonic stem cells
    Rangan Gupta
    Department of Veterinary Pathobiology, Bond Life Sciences Center, University of Missouri, Columbia, Missouri 65211, USA
    Mol Endocrinol 26:859-72. 2012
    ..Hence, a squelching mechanism underpins the transcriptional silencing of CGA by POU5F1 and could have general relevance to how pluripotency is maintained and how the trophoblast lineage emerges from pluripotent precursor cells...
  8. pmc Model systems for studying trophoblast differentiation from human pluripotent stem cells
    Toshihiko Ezashi
    Division of Animal Sciences and Bond Life Sciences Center, University of Missouri Columbia, 65211, USA
    Cell Tissue Res 349:809-24. 2012
    ....
  9. pmc Preeclampsia: multiple approaches for a multifactorial disease
    Kathleen A Pennington
    Department of Obstetrics, Gynecology and Women s Health, University of Missouri School of Medicine, 500 North Keene Street, Columbia, MO 65201, USA
    Dis Model Mech 5:9-18. 2012
    ..There remains a pressing need for novel approaches, new disease models and innovative investigators to effectively tackle this complex and devastating disorder...
  10. ncbi The contrasting effects of ad libitum and restricted feeding of a diet very high in saturated fats on sex ratio and metabolic hormones in mice
    Andrei P Alexenko
    Department of Animal Sciences, University of Missouri Columbia, Christopher S Bond Life Sciences Center, Columbia, Missouri 65211 7310, USA
    Biol Reprod 77:599-604. 2007
    ..Since males fed the VHF diet had neither more Y-sperm nor sired more sons than daughters, the dietary effects are manifested exclusively through the female...
  11. ncbi Rapid evolution of the trophoblast kunitz domain proteins (TKDPs)-a multigene family in ruminant ungulates
    Anindita Chakrabarty
    Department of Veterinary Pathobiology, University of Missouri Columbia, Columbia, Missouri 65211, USA
    J Mol Evol 63:274-82. 2006
    ....
  12. ncbi Origin and evolution of the TKDP gene family
    Anindita Chakrabarty
    Department of Veterinary Pathobiology, University of Missouri Columbia, Columbia, MO 65211, USA
    Gene 373:35-43. 2006
    ..The TKDPs provide an example of a gene family that is evolving rapidly by whole gene duplication, intragenic duplication events, and selection for amino acid (aa) change...
  13. pmc Engraftment of human iPS cells and allogeneic porcine cells into pigs with inactivated RAG2 and accompanying severe combined immunodeficiency
    Kiho Lee
    Division of Animal Science
    Proc Natl Acad Sci U S A 111:7260-5. 2014
    ..The pigs also tolerated grafts of allogeneic porcine trophoblast stem cells. These SCID pigs should have a variety of uses in transplantation biology. ..

Research Grants30

  1. CARDIOVASCULAR DYNAMICS AND THEIR CONTROL
    John E Hall; Fiscal Year: 2013
    ..End of Abstract) ..