Off-campus WSU users: To download campus access dissertations, please use the following link to log into our proxy server with your WSU access ID and password, then click the "Off-campus Download" button below.
Non-WSU users: Please talk to your librarian about requesting this dissertation through interlibrary loan.
Date of Award
D. Randall Armant
HBEGF, is present in the uterus at the time of embryo implantation and protects first
trimester TB cells from apoptosis and promotes their invasion. The hypertensive disease, PE, in
which TB invasion of the uterine arteries is reduced and TB apoptosis is elevated, is
characterized by a reduction in HBEGF expression. In this study using a first trimester cell line
and villous explant culture key components involved in HBEGF survival signaling pathway were
identified. Specific MMP inhibitors established the requirement for MMP2 in HBEGF shedding
and upregulation. NGS identified a HIF regulated gene, HSPA6 (HSP70B’) and using specific
inhibitors it was established that HSP70 regulates MMP2 mediated shedding of HBEGF at low
O2 and is functional upstream of MAPKs signaling cascade. To further investigate HBEGF
upregulation at low O2 using NGS and siRNA knockdown of DGCR8 it was demonstrated that
other components of RISC may be involved in regulation of HBEGF mRNA translation. These
findings suggest that trophoblast survival during early pregnancy requires this signaling pathway
and disruption of any component could lead to placental insufficiency. However, a global
platform is needed to study the pathophysiology of trophoblast cells and their role in placental
TRIC is an innovative platform to noninvasively acquire fetal cells. DNA was extracted
from fetal cells isolated from 20 specimens with gestational age as early at 5 weeks to 19 weeks.
This was followed by targeted sequencing using the Forenseq (Illumina) platform to identify
informative SNPs. Using the Forenseq software 89% ± 12% of the 94 SNPs were called
correctly in the fetal samples. Using the STR analysis out of the 20 fetal samples, 9 males were
confirmed. Therefore, TRIC not only provides fetal cells but also opens new venues of perinatal
TRIC provides ample RNA from isolated fetal cells for extensive transcriptomic analysis.
Using qPCR it was demonstrated that the fetal cells have higher expression for EVT specific
markers such as HLA-G and KRT7 and markers for invasion/migration such as CDH5 and
MMP9, whereas the maternal cells have a higher expression for epithelial markers such as CDH1
and ITGA6. This suggests that not only are the fetal cells EVT like but may also undergo an
epithelial mesenchymal transition. Comparison of fetal and maternal cells from normal group
using NGS identified 409 genes, of which top 5 upregulated and downregulated genes were
validated by qPCR. Pathway analysis of the differentially regulated identified pathways related
to invasion/migration, proliferation and differentiation, further suggesting the EVT like
expression of the fetal cells. NGS revealed 348 differential expressed genes on comparison of
the EVT like fetal cells from the EPL group to those from the normal group that were further
validated by qPCR. Bioinformatic analysis identified pathways related to apoptosis,
inflammation and placental disease.
The exact origin and biology of cervical EVT cells, and their relationship to the human
placenta and pregnancy outcomes, are widely unknown. Based on our study so far it can be
hypothesized that as the placenta grows, EVT cells are naturally shed into the cervical canal, and
their overall phenotype is comparable to the EVT cells residing in the placenta. These studies
will provide a clearer view of the utility of EVT cells obtained by TRIC from ongoing
pregnancies for investigating early placentation and mechanisms of pathology, and their potential
as a platform for prenatal tests to predict maternal risk of placenta-based disease.
Jain, Chandni V., "Molecular Regulation Of Trophoblast Survival During Placentation And Pathologies Of Placental Insufficiency" (2016). Wayne State University Dissertations. 1642.