USF Health researchers applied CRISPR technology to study the very large human non-protein coding gene expressed only in placenta, stem cells and certain cancers

TAMPA, Fla (March 16, 2020) — The placenta, an organ which attaches to the lining of the uterus during pregnancy, supplies maternal oxygen and nutrients to the growing fetus. Abnormal formation and growth of the placenta is considered an underlying cause of various pregnancy complications such as miscarriages, stillbirth, preeclampsia and fetal growth restriction. Yet, much remains to be learned about molecular mechanisms regulating development of this blood-vessel rich organ so vital to the health of a pregnant woman and her developing fetus.

Hana Totary-Jain, PhD, an associate professor of molecular pharmacology and physiology in the USF Health Morsani College of Medicine, was senior author of the study published in Scientific Reports.

University of South Florida Health (USF Health) Morsani College of Medicine researchers recently discovered how a very large human non-protein coding gene regulates epithelial-to-mesenchymal transition (EMT) – a process that contributes to placental development during early pregnancy, but can also promote cancer progression.

During the first trimester, fetal-derived placental cells known as trophoblasts invade the maternal uterine lining and modify its blood vessels to allow oxygenated blood to flow from the mother to fetus. However, trophoblast invasion requires tight regulation of EMT. If inadequate, trophoblast invasion is too shallow to adequately remodel the maternal blood vessels, and adverse pregnancy outcomes can occur. Conversely, excess EMT can cause exaggerated trophoblast invasion through the uterine wall leading to placenta accreta, a condition that can cause hemorrhage and often requires hysterectomy at delivery.

The USF Health researchers used a powerful genome editing technology called CRISPR (shorthand for “CRISPR-dCas9) to activate all of the chromosome 19 microRNA cluster (known as C19MC), so they could study the gene’s function in early pregnancy. C19MC — one of the largest microRNA gene clusters in the human genome — is normally turned off but becomes expressed only in the placenta, embryonic stem cells and certain cancers.

Dr. Totary-Jain discusses the molecular aspects of placenta development and pregnancy complications with research collaborator Umit Kayisli, PhD, a professor of obstetrics and gynecology at USF Health.

In their cell model study, published Feb. 20 in Scientific Reports, a Nature research journal, the USF Health team showed that robust activation of C19MC inhibited EMT gene expression, which has been shown to reduce trophoblast invasion.

But when trophoblast-like cells were exposed to hypoxia – a lack of oxygen similar to that occurring in early placental development — C19MC expression was significantly reduced, the researchers found. The loss of C19MC function causes differentiation of trophoblasts from stem-like epithelial cells into mesenchymal-like cells that can migrate and invade much like metastatic tumors. This EMT process helps explain trophoblast invasion and early placental formation.

“We were the first to use CRISPR to efficiently activate the entire gene, not just a few regions of this huge gene, in human cell lines,” said the paper’s senior author Hana Totary-Jain, PhD, an associate professor in the Department of Molecular Pharmacology and Physiology, USF Health Morsani College of Medicine. “Our study indicates C19MC plays a key role in regulating many genes important in early implantation and placental development and function. The regulation of these genes is critical for proper fetal growth.”

Above: Chromosome 19 microRNA cluster (stained purple) expressed in first-trimester placenta.  Below: In preparation for pregnancy, fetal trophoblast cells (brown) from which the placenta arises invade maternal decidual cells (pink) in the uterus lining. | Images courtesy of Hana Totary-Jain, originally published in Scientific Reports: doi.org/10.1038/s41598-020-59812-8

“You need EMT, but at some point the process needs to cease to prevent adverse pregnancy outcomes,” Dr. Totary-Jain said. “You really need a balance between not enough invasion and too much invasion, and C19MC is important in maintaining that balance.”

Dr. Totary-Jain and others in her department collaborated with colleagues in the medical college’s Department of Obstetrics and Gynecology on the project.

“The USF Health study offers new insight into how trophoblasts interact with the maternal uterine environment to become more invasive or less invasive in the formation of the placenta,” said coauthor Umit Kayisli, PhD, a USF Health professor of Obstetrics and Gynecology. “More research on microRNA expression and how it inhibits EMT may help us better understand the causes and potential prevention of preeclampsia and fetal growth restriction, which account for 5-to-10 percent of all pregnancy complications as well as spontaneous preterm births.”

Investigating the effects of altered C19MC expression on cell differentiation and trophoblast invasion has implications not only for a better understanding of normal and abnormal placental development, but also for cancer and stem cell research, Dr. Totary-Jain added.

Dr. Totary-Jain and Dr. Kayisli

Photos by Freddie Coleman, USF Health Communications and Marketing

The USF Health Department of Obstetrics and Gynecology was one of eight programs across the country selected to be highlighted in a television film series produced by the American Congress of Obstetricians and Gynecologists (ACOG).

Dr. Palmer with USF residents.

The finished film was premiered at ACOG’s 2017 Annual Clinical and Scientific Meeting, held May 7 to 9 in Chicago.

USF Health areas that were highlighted in the video, along with a grateful patient family, included:

• Residency Wellness Program – administers wellness modalities directly to the residents, including yoga, meditation, life-mapping, cranio-sacral therapy, music therapy, and more. Our residents tap into self-healing during and after these sessions, with the goal of teaching their patients these wellness techniques.
• Human Placenta Project –Anthony Odibo, MD, MSCE, FRCOG, FACOG, and Umit Kayisli, MSc, PhD, are co-principal investigators for the Human Placenta Project study, which is designed to predict poor growth of the fetus earlier in pregnancy so that physicians can intervene sooner to help prevent stillbirth, Cesarean delivery, decreased oxygen levels and other adverse outcomes. Our study is as one of 19 projects funded by the Human Placenta Project — an initiative launched by the NIH’s Eunice Kennedy Shriver National Institute of Child Health and Development to better understand the role of the placenta in health and disease.
• Twin-to-twin transfusion syndrome – Our Fetal Care Center of Tampa Bay is the leading fetal care center in Florida and the South East region, and remains among the most successful in treating twin-to-twin transfusion syndrome.
• Pregnancy Loss Prevention Center – its mission is to understand and help women who have experienced multiple miscarriages, stillbirths or premature births or the risk factors leading to them – from preeclampsia, diabetes and lupus to inflammation and infection.
• Simulated residency training at CAMLS – offers residents to participate in a variety of GYN, robotic, and obstetric simulations.

Dr. Anthony Odibo and Dr. Umit Kayisli pose for a portrait at USF Health’s South Tampa Center during the filming of 2017 ACOG video.

The finished film was broadcast on screens in high visibility areas throughout the conference and also in the ACOG TV programs. It will also remain live on ACOG TV website for a year. Here’s the link to the video, and here’s a link to the ACOG TV playlist.

Behind-the-scenes glimpse at USF Health Ob/Gyn resident director Dr. James Palmer during the filming of the 2017 ACOG video.

Photos by Sandra C. Roa, USF Communications

Bridging laboratory and clinical sciences, the study aims to improve the health outcomes  of pregnancies complicated by poor fetal growth

Tampa, FL (Sept. 28, 2015) – The USF Health Morsani College of Medicine has received a $4-million National Institutes of Health grant that will employ new imaging technologies and test biomarkers in the blood to determine whether abnormalities in the smallest blood vessels of the placenta and negative environmental influences, particularly smoking, cause fetal growth restriction (FGR).

The ultimate goal of the four-year study is to design a reliable way to predict poor growth of the fetus earlier in pregnancy so that physicians can intervene sooner to help prevent stillbirth, Cesarean delivery, decreased oxygen levels and other adverse outcomes.

The USF research award (1U01HD087213-01) was announced today as one of 19 projects funded by the Human Placenta Project — an initiative launched by the NIH’s Eunice Kennedy Shriver National Institute of Child Health and Development to better understand the role of the placenta in health and disease.

Anthony Odibo, MD (left) and Umit Kayisli, PhD, of the USF Health Department of Obstetrics and Gynecology, are co-principal investigators of a $4-million Human Placenta Project — one of 19 new projects awarded in the U.S. and Canada by the NIH.

“I am so proud of our team,” said Charles J. Lockwood, MD,  dean of the USF Health Morsani College of Medicine and senior vice president for USF Health. “This is an important NIH initiative which addresses the common source of most major adverse pregnancy events – abnormal placentation.”

“In the past, it has been challenging to identify which women may benefit (from early therapeutic intervention), because they are at high risk for fetal growth restriction,” said co-principal investigator Anthony Odibo, MD, professor in the USF Department of Obstetrics and Gynecology.  “But powerful new imaging technology gives us the opportunity to really visualize all the blood vessels in the placenta, study the underlying problem of growth restriction, and create a highly predictive model for identifying small babies at risk of FGR.”

The USF grant, bridging laboratory and clinical sciences, will be led by Dr. Odibo and co-principal investigator Umit Kayisli, PhD, associate professor of obstetrics and gynecology.  Dr. Odibo, specializing in maternal-fetal medicine, is an expert in fetal therapy and directs the USF Fetal Care Center at Tampa General Hospital.  Dr. Kayisli specializes in molecular and cellular biology in reproduction and its clinical implications.

They will work on the NIH project with USF Ob/Gyn co-investigators Charles J. Lockwood, MD, Frederick Schatz, PhD, and Ozlem Guzeloglu-Kayisli, PhD, and with Rajendra Kedar, MD, from the USF Department of Radiology.  USF colleagues at Necker Hospital in Paris and at Oakland University William Beaumont School of Medicine in Rochester, MI, will also collaborate.

Fetal growth restriction (FGR), affecting up to 10 percent of all pregnancies, is commonly defined as fetal weight below the 10^th percentile for gestational age as determined by ultrasound. The condition remains a leading contributor worldwide to the death and illness of babies before and after birth.

Placental function – the ability of the critical organ to shuttle blood, oxygen and nutrients from mother to fetus through an intricate network of blood vessels – is inadequate in pregnancies complicated by FGR.  But predicting FGR has been difficult, because until recently imaging technologies have not been sensitive nor specific enough to clearly detect the smallest blood vessels in the placenta and monitor the flow of blood through this branching microvasculature.

Dr. Odibo points to an ultrasound image of the placenta, a critical organ that shuttles blood, oxygen and nutrients from mother to fetus through an intricate network of blood vessels.

For the USF study, researchers will use two of the latest technologies – superb microvascular imaging, or SMI ultrasound, and blood oxygen level-dependent magnetic resonance imaging, or BOLD MRI.

The investigators will compare biopsies of placenta from normal and FGR-complicated pregnancies in the laboratory and correlate them with the imaging assessments of the placental microvasculature.  They will also study how smoking affects the microvasculature and the potential link with FGR.

“The results obtained from SMI ultrasound and BOLD MRI combined with changes in expression levels of several biomarkers and epigenetic modifications in response to smoking will be instrumental in developing a predictive model for pregnancies at high risk for fetal growth restriction and improving the sensitivity and specificity of a potential early diagnosis and treatment of FGR,” Dr. Kayisli said.

For a list of all new grants awarded as part of the NIH Human Placenta Project, go to http://www.nichd.nih.gov/news/releases/Pages/092815-NIH-awards-HPP.aspx.

-USF Health-

USF Health’s mission is to envision and implement the future of health. It is the partnership of the USF Health Morsani College of Medicine, the College of Nursing, the College of Public Health, the College of Pharmacy, the School of Physical Therapy and Rehabilitation Sciences, and the USF Physician’s Group. The University of South Florida is a Top 50 research university in total research expenditures among both public and private institutions nationwide, according to the National Science Foundation.  For more information, visit www.health.usf.edu

Media contact:
Anne DeLotto Baier, USF Health Communications
(813) 974-3303 or abaier@health.usf.edu

Photos by Eric Younghans, USF Health Communications and Marketing