Tag Archives: preeclampsia

Protecting the Miracle of Childbirth

The spectacular process of human reproduction is complex, time consuming, and, above all, fascinating! Much has been learned over the years dealing with the mechanisms of pregnancy in many of Earth’s lifeforms. The research on genetics, like uncovering the entire human genome, makes incredible strides toward fully grasping why certain physiological processes happen. However, there are still numerous question marks, specifically speaking about women’s health during pregnancy and after, that require research and understanding. Full efforts are being undertaken that aim to ultimately lead to safer pregnancies, better means of treating diseases, and developing new techniques.

Research Project

Preeclampsia is a disorder during pregnancy characterized by high blood pressure and excess protein excretion. Because the condition is not entirely understood, treatment options are far and few between for women suffering. Currently, the only remedy is a low-dose treatment of aspirin. The effects and mechanisms of this aspirin treatment is not completely understood either, so the purpose of my study is to attempt to demystify the workings of the treatment. Specifically, I am targeting human trophoblast cells, the major cell type involved in the development of the placenta, an organ that provides nutrition to the developing fetus. By varying different doses of aspirin, I am examining the changes, or lack of change, in the trophoblast DNA. If changes are observed, we will have knowledge on how and if aspirin will help women suffering from preeclampsia, which will ultimately lead to a safer pregnancy for both the mother and child.

Realities of Research

It has been an exciting experience working in a research lab. Not only have I learned valuable techniques, but I am directly impacting the future of medicine, even if it is in a seemingly small way. I have been surprised by the level of attention and precision that is addressed when conducting research. I always knew that attention to detail was important, but the extent to this precision that I have been performing has shocked me along with how these techniques were practiced. For example, RNA isolation is a delicate, yet simple process that requires attention and a good grasp on how to pipette well. If a step is skipped, such as forgetting to add the homogenate additive, then the RNA yield could be put at risk. It is too early in my research stages where results and conclusions can be made. Typically, one trial of cell growth requires one full week, so multiply one trial by the many that we are attempting and the overall experiment becomes lengthy.

Life of a Scientist

The day-to-day life of a young scientist has been exciting. While not all of the parts of my day are groundbreaking and entertaining, it is still a rewarding process. I usually begin my day with notebook entries, planning, and reading up on current events in my field. My research involves a fairly strict time schedule, so in the afternoon, the experimenting and cell ‘farming’ as I call it, can begin. I was surprised by the equipment that I have at my fingertips.  Nothing is more thrilling than looking through a $45,000 microscope or running a 6 well plate through a machine that you can’t even pronounce. The best part so far for me has been the adjusting to a real life laboratory. I have begun to entertain the idea of having my own lab in the future, and becoming familiar with how a lab is run has been a wonderful experience. The worst part has been the waiting that is required between experiments. It makes me wish I had a magic wand that would make the cells grow and be ready for testing at the flick of my wrist. It has been so wonderful working with everyone in my lab. I love the feeling of having an independent project, but still being under a larger umbrella of research with my coworkers where we can discuss information and findings.

 

Brandon Cooley is a junior at the University of Iowa where his is studying biology. His future plans involve graduation with his degree and enrolling in an MD/PhD program where he can further develop his researching skills while being present as a clinician in a hospital!
Novel Mechanisms of Preeclampsia Prevention via SGK1 and Corticosteroids

Preeclampsia, a hypertensive disorder in pregnancy, affects more than 6 million pregnancies per year worldwide. It is a dangerous condition during pregnancy which involves high blood pressure, proteinuria, and swelling. The Santillan lab has shown that single dose early of BMTZ early in pregnancy will reverse late pregnancy hypertension and proteinuria (1). The molecular mechanism by which this reversal occurs is unclear. One potential pathway involves serum/glucocorticoid regulated kinase 1 (SGK1), a serine/threonine kinase stimulated by corticosteroids. SGK1 dysregulation and human genetic variants in SGK1 have been associated with hypertension. The overall goal of this study is to determine whether SGK1 and its regulation play a role in preeclampsia.  If so, BMTZ has the potential to prevent preeclampsia in humans. Because mir-365 has been shown to decrease SGK-1 expression in human placentas from pregnancies with poor placentation, I will examine the effect of vasopressin and SGK1 in placental cells on mir-365 expression. In addition, it will be determined if placental mir-365a-3p is differentially expressed in human preeclampsia. This project is significant because it may help to determine how BMTZ protects from preeclampsia and whether BMTZ could be useful in humans.

 

Research in the lab can be very stressful. Things may not go as well as expected and troubleshooting is a process. Regardless, I had the opportunity to learn many new techniques that would help me in the future. I was surprised how research is comprised of so many different aspects. A little difference in one experiment may change the whole outcome. I learned a great set of skills like how to maintain a cell culture, perform an ELISA, BCA, and extract RNA. It took a while for me to start up on my experiments because I had to research some more background information to ensure I knew what I was doing. My experiments went smoothly, but it was later found that the drug I was using to treat the cells was not working in our mouse model; therefore, it may not be working with my cells as well. My project was put to a halt to first determine if the drug was correctly performing. The drug was aliquoted about a year ago and may have degraded. I would have to wait in order to determine whether I was able to continue or to start over. In the meantime, I worked with my mentor with small projects and learned useful techniques. Additionally, I worked on the second portion of my project involving whole placental tissues. The tissues were RNA prepped and analyzed via qPCR. The results showed that there was a significant difference with p-value of 0.016. This makes sense because Xu found that miR-365 negatively regulates IL-6 and it, in turn, is transcriptionally regulated by Sp1 and NF-κB. (2) So, transcriptional down-regulation of miR-365 should result in increased IL-6. This was interesting to hear, but we cell culture was needed to determine this and it was on standstill.

 

There were some busy days and other days there was a lot of down time. For example, one day there may be multiple tests to complete in a day, other days an experiment would consist of wait time. The most surprising part of participating in the lab is that I realized that a lot of the down time is used to write papers or grants. Research involves a great deal of writing to express the study to the public eye and document previous studies to help ongoing studies. I am appreciative of researchers because without those papers I would not have been able to understand my study without background information. Most days were very stressful trying to balance all of the work and trying to understand why a certain mechanism happened. My least favorite part during my time in the lab was working so hard on an experiment and in the end, not having it work out. The best part was working along with my mentor to learn new techniques and tests. I’m also glad that people around the lab worked well with one another and that they would take the time to reach out and teach me.

 

References:

  1. Santillan, M., Santillan, D., Scroggins, S., Min, J., Sandgren, J., Pearson, N., Leslie, K., Hunter, S., Zamba, G., Gibson-Corley, K. and Grobe, J. (2014). Vasopressin in Preeclampsia: A Novel Very Early Human Pregnancy Biomarker and Clinically Relevant Mouse Model. Hypertension, 64(4), pp.852-859.
  2. Xu et al. miR-365, a Novel Negative Regulator of Interleukin-6 Gene Expression, Is Cooperateively Regulated by Sp1 and NF-κB. Journal of Biochemistry 286: 21401-21412, 2011
Carolyn Lo is a junior majoring in Human Physiology and Biochemistry at the University of Iowa in Iowa City, Iowa. She is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working with Dr. Mark Santillan at the Carver College of Medicine in Iowa City, IA. Carolyn’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (NHLBI) (Grant #1 R25 HL115473-01). After graduation, Carolyn plans to pursue a doctorate degree in medicine.