Approximately every 38 seconds, an American will experience a myocardial infarction (MI). The prognosis after MI depends on the amount of myocardial death. Therefore, finding
cardioprotective therapies is an important field of research. From past research conducted in our laboratory, results were obtained to show that a protein called BMP3b, secreted by brown adipose tissue (BAT), protects mice hearts during myocardial infarctions.
This summer, we went a little further in our investigation by looking at how BMP3b affects angiogenesis, the development of new blood vessels from pre-existing blood vessels. It is already known that BMP3b modulates angiogenesis, but the exact mechanism on how this is done is rather unclear. This study was completed using human umbilical vein endothelial cells (HUVEC) in which half of the cells had BMP3b protein introduced to them and the other half had a saline solution introduced to serve as a control. Then half of the cells introduced to BMP3b and saline solution were introduced to normoxic conditions and the other half was introduced to hypoxic conditions. Normoxia would simulate normal heart conditions whereas hypoxia, a lack of oxygen, would mimic myocardial infarctions. Angiogenesis is stimulated by HIF1a, the most important transcription factor on hypoxia.
Using these four conditions, a qPCR was made to see how the presence of BMP3b protein influenced RNA expression of angiogenesis-related genes. The genes that were specifically looked at were ANGPT1, ANGPT2, THBD, and VEGF. GAPDH was also looked into as it served as a housekeeping gene for the experiment. Additionally, there was a chance that there would not be much variation in RNA expression amongst the four different conditions, so a proteome profiler human angiogenesis array kit was used to detect the relative levels of expression of 55 angiogenesis-related proteins without performing individual assays for each protein.
Through this research experience, I have learned how to utilize several new laboratory techniques. I feel more comfortable working with cell cultures. I have also gained some understanding on how to conduct qPCRS and how the process works. Additionally, I have learned how to make western blots, block membranes containing proteins, develop films in a dark room, and follow intricate protocols to accomplish a goal of an assay. My laboratory also allowed me to gain some experience in getting to work with mice which I found to be quite exciting and interesting. Many of the techniques that I have learned over this summer will definitely serve me some purpose in the future as I embark on my own research endeavors.
Working in my lab has also contributed to my educational experience as a premed student as it introduced me to the world of cardiology. Prior to this lab experience, I had not really considered the field for a future career. However, after working alongside with my post doc, Ingrid Marti, I have gained a deeper understanding and a newfound interest on the topic of myocardial infarctions. I really enjoyed my PURM experience.