hypoxia | APS Undergraduate Researcher

This summer I worked on a metabolomics project surrounding the effects that hypoxia, or deficient oxygenation, has on oxidative stress and inflammation. I used metabolomics, or the study of the functional molecules in the body, to interpret the molecular changes that eventually lead to physiological complications. Currently, we know that oxylipins, biological molecules that are metabolized from polyunsaturated fatty acids (PUFAs), are markers of oxidative stress and inflammation in conditions such as hypoxia (1). My lab extracted venous plasma from fetal and newborn sheep that were living at high-altitudes, as hypoxic conditions can be simulated by high-altitude living. By running tests that quantified the oxylipins in fetuses and newborns, I was able to distinguish which metabolites were prominently affected by hypoxic conditions. Later, I was able to find pathways, tracing how certain PUFAs were metabolized and from which PUFAs certain oxylipins were derived. Based on these relationships, I aimed to find possible roots for the oxidative stress and inflammation caused by chronic hypoxia.

These findings highlight our understanding of lipid metabolism as it is affected by high-altitude hypoxia. This study has the potential to help us develop treatments that target inflammatory pathways induced by pre and post-natal hypoxia. For instance, the enzymatic pathway CYP, which metabolizes PUFAs proved to play a large role in the production of oxylipins that were affected by hypoxia. Targeting this pathway early in the womb may help prevent lung dysfunction that may develop just after birth.

An area in my research that I found difficult was the dense literature. At first sight, it was intimidating‒ scientific jargon and compound nomenclature most of all. I realized that as I started connecting terms to function‒ associating oxylipins with potential roles in the body was now feasible. The truth is, it takes time and understanding to grasp the material, but the more I read and the more I searched, the less intimidating it all felt. Around the lab, there are several skills to master‒ several of which consist of success and failure. For instance, I had a hard time developing networks for my metabolites and working with statistical software early on in my research; this is now something I wish to improve. Often, I received results from my data that I did not expect and it reminded me how difficult it was to remain impartial. I ran into a list of questions that, over time, clarified and narrowed what in fact my research would delineate.

I could summarize the life of a scientist in one word: unpredictable. It surprised me how difficult consistency actually was with data. That being said, it is a huge task to filter data and focus on only a few aspects of it; everything seemed important. Moreover, a reliable, cooperative lab team is a vital component to a scientist’s life in the lab. While not everyone is specialized in the same subject or project, a team creates a supportive environment where we feed off of one another’s knowledge and work in collaboration for the interest of science.

References

Gabbs M, Leng S, Devassy JG, Monirujjaman M, Aukema HM. Advances in Our Understanding of Oxylipins Derived from Dietary PUFAs. Adv Nutr 6: 513–540, 2015.

Vanessa Lopez is a junior Biochemistry major at Occidental College in Los Angeles, CA. She is a 2017 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow. She works with Dr. Sean Wilson in his lab at Loma Linda University in Loma Linda, CA. Vanessa’s fellowship is funded by the APS, as well as a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). Her research is also supported by NIH grants HD083132 [LZ], 1U24DK097154 [OF] through Dr. Wilson’s lab. She is interested in endocrinology and dietetics. Her plan is to go to medical school after graduation.

My research project this summer focuses on the development of rehabilitative strategies used to combat respiratory dysfunction (the inability to breathe) in patients suffering from spinal cord injuries. Following a spinal cord injury, brainstem projections traveling down the spinal cord are severed, ultimately leading to paralysis of respiratory muscles such as the diaphragm. Consequently, breathing impairments are the primary cause of death for spinally-injured patients. This summer project focuses on a rehabilitative therapy known as intermittent hypoxia (IH) which involves brief and repeated exposures to low oxygen levels over a period of time. When given in low doses (>15 episodes/day), intermittent hypoxia can improve leg strength, walking function, and respiratory function in spinally injured rats and humans. On the other hand, high doses of intermittent hypoxia can elicit spinal inflammation and other pathological consequences. My study seeks to determine whether activation of a certain protein kinase that acts as a marker of inflammation, p38 MAP kinase, is increased in all dose protocols of intermittent hypoxia or just high dose intermittent hypoxia. Assessing the impact of p38 MAP kinase phosphorylation/activation is crucial for the development of safe and effective methods to restore respiratory function in individuals with a spinal cord injury. Currently my research subjects are rats; however, once our lab can ensure that low dose intermittent hypoxia is safe and non-harmful to humans, through my project and many others currently being conducted, we can transition our treatments from animals to humans.

What was it like doing research in a lab?

My research experience this summer was an eye-opening experience that, for the first time in my professional career, allowed me to devote all my time to the research I am passionate about. Conducting research in the lab alongside other undergraduate and graduate students, and post-doctoral fellows, was nerve-wracking yet very rewarding. When I first began my project, part of me was afraid to attempt tasks and techniques that were new to me for fear of ruining valuable data. However, to my surprise everyone in the lab was eager to help me in any way they could even if they were occupied with their own projects. I quickly learned that I would have to develop several techniques for my project to be a more independent and productive researcher. These techniques include immunohistochemistry (IHC) staining and imaging. Thus far, my experiment has been working, with all CTB injections, exposure treatments, perfusions, harvesting, and tissue sectioning going successfully. An unexpected mishap that occurred involved the immunohistochemistry staining. During our first IHC staining trial, we stained only a few of the tissues and had difficulty viewing the markers that were stained for when imaging. This was a result of having used an antibody at a wavelength of light that wasn’t appearing on the microscope. After reviewing literature and past protocols we decided on a different antibody that would produce better images. Although unfortunate, this setback demonstrated that while conducting research several things can go wrong but can just as easily be fixed. As for results, my project is still under the works as I have just reached the imaging phase.

I have found my experience as a scientist this summer to be both rewarding and stimulating. Every day when I come into lab, I am eager to jump into my project and engage in conversation with other researchers about the impact all of our research could one day have. My days consist of running experiments, performing animal care, cutting tissue, and whenever I have down time, reading papers. The best part of my research experience is collaborating with other members in the lab. I enjoy listening to the advice and thoughts of the other researchers because it gives me other perspectives on things. The worst part about being a research scientist is probably accepting mistakes and figuring out how to solve the problems. I found working as part of the team to be an educational experience that helped me improve my communication and leadership skills.

Juliet Santiago is a junior majoring in Microbiology and Cell Science at the University of Florida in Gainesville, FL. She is a 2017 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Gordon Mitchell’s lab at the University of Florida in Gainesville, Florida. Juliet’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduation, Juliet plans to pursue a career as a biomedical scientist in industry or government.