Placental and endometrial donations

The Loke Centre for Trophoblast Research (CTR) promotes research into maternal-fetal interactions during normal pregnancy and how complications, such as infertility, miscarriage, restricted growth of the baby and pre-eclampsia, arise. Women’s health is an understudied area, which causes considerable ill-health and distress. Pregnancy-related complications and disorders account for much of this burden, causing significant maternal and fetal maternal morbidity and mortality, and having life-long effects on the mother and her offspring. The World Health Organisation estimates 800 women die worldwide each day due to issues around pregnancy and childbirth. Pre-term birth complications, birth asphyxia, birth trauma and neonatal sepsis and infections account for 8% of global disease burden (WHO). The Loke CTR brings together researchers interested in different aspects of reproduction and pregnancy, from basic science to clinically related projects. It also includes clinicians and scientists working on virally-induced cervical and endometrial cancer, and on conditions such as endometriosis.

The Loke CTR Biology of the Human Uterus in Pregnancy and Disease Biobank, REC reference number 22/EE/0133 facilitates bringing together both non-pregnant and pregnant maternal endometrial samples from the womb, and placental samples at different stages of gestation. Specifically, our tissue collection includes:

• First trimester samples from women undergoing terminations of pregnancy
• Term placental tissue samples from healthy and pathological pregnancies
• Endometrial scratch biopsies form patients undergoing fertility treatments

All samples are collected on an anonymised basis. The tissue is utilised to address questions pertaining to reproductive health. The principal focus of the Loke CTR research is the establishment of a normal pregnancy and the origins of complications, such as miscarriage, poor fetal growth and pre-eclampsia. We are also interested in how common disorders of the endometrium, the lining of the womb, may arise, such as endometriosis and endometrial and cervical cancers.

The research takes a basic science, laboratory-based approach, involving experimental manipulations of cell cultures, or analyses of gene activity or cellular signalling networks in different tissues or cell isolates. We are interested ultimately in translating our findings to improving clinical outcomes. To this end, the research is based on samples from clinically well-phenotyped groups of women; for example, we investigate molecular and genetic markers of endometrial function in women with infertility or recurrent miscarriage. We also investigate how the placental molecular pathology differs in sub-types of pre-eclampsia, shedding light on the origins of this syndrome with the hope of identifying new avenues for intervention or treatment.

Whilst the majority of the data produced is relevant to the discipline of women's health, it is possible that some of the more basic research may have implications for the wider bio-medical field; for example, knowledge of the transcriptional networks maintaining trophoblast or endometrial stem cells may be relevant to other cell types. Equally, understanding the immune interactions at the maternal-fetal interface between genetically-related, but different, individuals may provide new insights into the evolutionary selective pressures that have shaped our immune system.

Examples of research projects supported by placental and endometrial donations:

• Pregnancy at high-altitude and maintenance of placental energy levels, Andrew Murray

At high altitude, oxygen delivery to the placenta and developing baby is challenged. This can result in lower birthweights and increases the risk of severe health problems for pregnant mothers and their babies. In human populations that have lived at high altitude (e.g. in La Paz, Bolivia) for many thousands of years, there is relative protection against pregnancy complications, and understanding this protection may hold the key to helping some pregnancy complications at sea level. This project focuses on the impact of high altitude on placental mitochondria – the parts of the cell that consume oxygen for energy metabolism. To do this we study placentas from sea level and high-altitude regions in Colorado, United States and La Paz, Bolivia.

• In vitro treatment of inflammatory cytokines and high fat high sugar conditions on human primary trophoblast nutrient uptake and growth signalling, Amanda Sferruzzi-Perri 

Primary cytotrophoblasts are isolated from human term placenta using a well-established technique using sequential trypsin and DNAse digestion followed by Percoll gradient purification. Cytotrophoblasts are then left in culture to spontaneously differentiate until hour 72. From hour 72, treatments such as inflammatory cytokines (IL1α, IL6, TNFβ), high fat (palmitic acid), high sugar (glucose), and lipopolysaccharide (LPS) are administered for 24 hours. At hour 96, glucose and fatty acid uptake assays are performed, and cells are lysed for protein and RNA extractions. These protein and RNA lysates will probe for factors involved in inflammation, nutrient transport, and growth signalling.

• What is the global DNA methylation patterning in choriocarcinomas compared to healthy placental cells? Courtney Hanna

Our genes can be switched on or off by small chemical changes to DNA, known as DNA methylation. This process is very important during development, helping cells to grow and take on their proper roles. The placenta has a distinctive DNA pattern compared to other organs, with many genes only partly switched off.  Interestingly, this unusual pattern is also found in many cancers. In very rare cases, placental cells can change and form a type of cancer called choriocarcinoma, which is highly invasive and dangerous. At the moment, we do not fully understand why this happens.
In this project, we will study DNA from both choriocarcinoma tumour samples and healthy placental tissue. Using advanced DNA technique called whole genome bisulphite sequencing, we will look at the chemical ‘on/off switches’ across the whole genome. By comparing the tumour tissue with healthy placenta, as well as with data from other types of placental cells, we hope to identify the specific DNA changes that may drive this cancer. This research will not only help us understand why this rare cancer sometimes arises, but may also improve wider knowledge about pregnancy health and cancer development.

• The impact of obesity and diabetes during pregnancy on placental energy production,  Andrew Murray

Globally, there are an increasing number of pregnancies where one or both parents have obesity and/or diabetes. These conditions can alter the delivery of nutrients to the placenta and the developing baby, and can have longer term consequences for the health of the mother and baby. In this project, we are aiming to understand how maternal metabolic health impacts on the placental mitochondria – the parts of the cell responsible for energy metabolism – and the consequences for longer-term infant and maternal health.