Prenatal diagnosis using cells obtained from the developing conceptus can provide critical information about obstetric pathophysiology and fetal genetics. This information can be utilized to anticipate genetic disorders brought about by heredity or chromosomal errors that increase with maternal age. The most common methods of retrieving fetal tissue for prenatal diagnosis, amniocentesis and chorionic villous sampling, are invasive, with a risk of fetal loss. Additionally, they are not available before 9-14 weeks of pregnancy. However, fetal trophoblast cells migrate from the placenta into the cervix as early as 3 to 4 weeks after fertilization, when most pregnancies are first detected. The proposed investigation will build upon past success in refining a minimally invasive method to recover fetal cell from the cervix that is essentially identical to a PAP test. We can reliably collect and identfy trophoblast cells in specimens obtained by transcervical sampling (TCS) from pregnant women early in the first trimester. Furthermore, we can isolate the trophoblast cells free from resident maternal cervical cells by immunomagnetic affinity separation to be used for genetic analysis of fetal DNA.
In Specific Aim 1, the fetal DNA will be validated by short tandem repeat (STR) profiling and comparison to blood samples from the mother and corresponding neonate (N=50). Single cell approaches adapted from preimplantation genetic screening will identify individual fetal cells in heterogeneous isolates. Microgram quantities of DNA from individual cells generated by whole genome amplification (WGA) will be used for STR profiling and saved for further genetic analysis. Structural chromosomal anomalies cause birth defects and are a high priority for prenatal genetic testing.
In Specific Aim 2, the utility of trophoblast cells obtainedby TCS for chromosomal analysis will be examined by considering the hypothesis that these placenta-derived cells accurately report the status of chromosomes in those fetal cells that develop to form the neonate. Amplified fetal DNA from samples studied in Aim 1 will be compared to neonatal DNA by comparative genomic hybridization to determine the numerical status of all 24 human chromosomes. These studies will establish the limitations of TCS for providing reliable genetic information on the fetus during the first trimester. The methodologies developed in this investigation could eventually establish clinical tests that use TCS, providing information on the genetic status of the fetus before development has progressed beyond the first two months.

Public Health Relevance

This research could provide major public health benefits by introducing a safe, minimally invasive method to capture fetal cells at the beginning of pregnancy for cellular and genetic study. The utility of this procedure for chromosome analysis will be investigated to develop prenatal genetic diagnostic tests that would provide immense opportunities for improving the health and well-being of mothers and their babies.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21HD071408-02
Application #
8508086
Study Section
Pregnancy and Neonatology Study Section (PN)
Program Officer
Yoshinaga, Koji
Project Start
2012-08-01
Project End
2014-07-31
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
2
Fiscal Year
2013
Total Cost
$216,372
Indirect Cost
$74,022
Name
Wayne State University
Department
Obstetrics & Gynecology
Type
Schools of Medicine
DUNS #
001962224
City
Detroit
State
MI
Country
United States
Zip Code
48202
Bolnick, Jay M; Kohan-Ghadr, Hamid-Reza; Fritz, Rani et al. (2016) Altered Biomarkers in Trophoblast Cells Obtained Noninvasively Prior to Clinical Manifestation of Perinatal Disease. Sci Rep 6:32382
Armant, D R; Fritz, R; Kilburn, B A et al. (2015) Reduced expression of the epidermal growth factor signaling system in preeclampsia. Placenta 36:270-8
Fritz, Rani; Kohan-Ghadr, Hamid Reza; Sacher, Alex et al. (2015) Trophoblast retrieval and isolation from the cervix (TRIC) is unaffected by early gestational age or maternal obesity. Prenat Diagn 35:1218-22
Armant, D Randall (2015) Intracellular Ca2+ signaling and preimplantation development. Adv Exp Med Biol 843:151-71
Fritz, Rani; Kohan-Ghadr, Hamid-Reza; Bolnick, Jay M et al. (2015) Noninvasive detection of trophoblast protein signatures linked to early pregnancy loss using trophoblast retrieval and isolation from the cervix (TRIC). Fertil Steril 104:339-46.e4
Bolnick, Jay M; Kilburn, Brian A; Bolnick, Alan D et al. (2015) Sildenafil stimulates human trophoblast invasion through nitric oxide and guanosine 3',5'-cyclic monophosphate signaling. Fertil Steril 103:1587-95.e1-2
Bolnick, Jay M; Kilburn, Brian A; Bolnick, Alan D et al. (2015) Sildenafil Prevents Apoptosis of Human First-Trimester Trophoblast Cells Exposed to Oxidative Stress: Possible Role for Nitric Oxide Activation of 3',5'-cyclic Guanosine Monophosphate Signaling. Reprod Sci 22:718-24
Patel, Shivali; Kilburn, Brian; Imudia, Anthony et al. (2015) Estradiol Elicits Proapoptotic and Antiproliferative Effects in Human Trophoblast Cells. Biol Reprod 93:74
Bolnick, Jay M; Karana, Rita; Chiang, Po J et al. (2014) Apoptosis of alcohol-exposed human placental cytotrophoblast cells is downstream of intracellular calcium signaling. Alcohol Clin Exp Res 38:1646-53
Tiruvannamalai-Annamalai, Ramkumar; Armant, David Randall; Matthew, Howard W T (2014) A glycosaminoglycan based, modular tissue scaffold system for rapid assembly of perfusable, high cell density, engineered tissues. PLoS One 9:e84287

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