Internship

Project summary

Stiffening of the liver, better known as liver fibrosis, is caused by the formation of large amounts of scar tissue. This scar tissue is formed as a result of liver injury, which can be caused by metabolic disorders, obesity, alcohol use, drug use, or viral infection. Hepatic stellate cells (HSCs) will become activated due to this injury and drive fibrogenesis in order to heal the wounded liver. In the case of liver fibrosis, the injury is chronic, and HSCs are persistently activated, leading to the buildup of scar tissue and eventually cirrhosis. Because of the limited understanding of the mechanisms driving this, there are no FDA approved treatment options that will limit or reserve fibrosis. Fibrogenesis is driven by increased transcription and translation of extracellular matrix proteins by HSCs. This increased translational demand places a stress on HSCs, which activates the unfolded protein response (UPR). One branch of the UPR that regulates translation is PERK signaling. We have found that PERK signaling increases with HSC activation. We are engineering a PERK KO cell line using CRISPR-Cas9 technology in order to determine if PERK signaling is crucial to HSC activation and fibrogenesis. 

About the Internship

Dr. Jessica Maiers is studying the role of hepatic stellate cells in liver fibrosis, or the stiffening of the liver due to overexpression of protein involved in liver regeneration. The endoplasmic reticulum, an organelle tasked with folding proteins, plays an important role in fibrosis, but the role it plays is different between hepatocytes (regular liver cells) and hepatic stellate cells (HSCs), the cells that become activated in liver regeneration. Because liver fibrosis is caused by the overexpression of proteins, understanding where those proteins are coming from and the pathways they take in HSCs is crucial.

Workplace

Some of my strengths that I've noticed so far have been flexibility and fast-learning. Now that I have LHSI added to my list of responsibilities, I've had to rearrange my usual daily routine and make more room on my schedule. This has forced me to keep open spots on my schedule while still keeping the important things within a reasonable time. For example, I typically go to my LHSI site on Tuesdays at 3:15; I would usually use this time to run errands and get assignments done, but now I have moved this time to Wednesday morning or afternoon. This is a time that still works for me while allowing me to attend my internship. Having to be flexible has also given me a good idea of where I can open up my schedule in the future if need be. Fast-learning is another strength I have been able to work on. I am unfamiliar with most of the lab procedures, but I do not have much time to spend dwelling over them. I have learned to jump into new things without fear, and because of this, I've quickly learned many basic procedures!

I hope to become more familiar with the lab; not just the technical aspect, but the social aspect as well. Perfecting your lab techniques is just as important as getting along with your coworkers. I hope to familiarize myself with the lab culture and etiquette so that any teamwork can flow smoothly. Being respectful and professional is key to producing great work, and this is especially important in research.

At the beginning, integrating myself into the lab team has been difficult, but as time went on, I found a role for myself. I started off as an observer; simply watching others in the lab and letting them explain their work to me. It wasn't long before I started doing more hands-on work, and instead of observing, I was experimenting. Not quite doing a whole lot of work yet, but I was allowed to do simple things such as pipetting, checking on cells, counting cells, etc. And soon, when I got a better hang of these things, I began helping out with real experiments. Instead of just going through the motions of the work, I would collect results and discuss them. I created figures out of results from experiments and analyzed them to understand what I was looking at. Outside of the main experiments I help out with, I also help with general housekeeping, such as changing cell media or splitting cell plates when needed. These smaller tasks are important because they keep the cells alive, which is obviously important for experiments. Having these cell lines provides quick and easy access to cells whenever a new experiment will be done. These routine tasks I do keeps this work out of the hands of other teammates, and it also keeps them from having to retrieve frozen cells whenever a new experiment must be done. This along with the bigger work I do has become routine for me, and this has helped me solidify my role as a team member and keep me motivated.

Another benefit of teamwork is the diversity of each team member. Having different perspectives is especially important in science; it's what allows for new ideas or discoveries. Being surrounded by people with diverse backgrounds and though processes, I've been able to see things through a different lens. Having difficult concepts put into a new perspective has helped me see them at all angles, and I've been able to ask the right questions and make thorough analyses of new information or data.

Successes and Challenges

One success I've had so far is learning the steps of a Western blot (a technique used to visualize protein amounts in a sample of cells) from start to finish, from collecting cells and quantifying proteins to developing the blots. I've been learning the steps in chunks for a while, and with each part I've learned, I've spent time practicing it. Recently, I finally learned the last part I needed to know and putting it all together was very satisfying. On the other hand, a challenge I've faced during the internship was understanding the mechanisms behind lab techniques. Knowing why a certain buffer or reagent is needed can be important to performing an experiment correctly. For example, in the past while doing a Western blot, I used a different type of membrane, and this new type of membrane needed to be treated with methanol before use. I had forgotten this step, and I ended up having to restart the blot. Another worker in the lab explained to me why one membrane needs to be treated with methanol and why one doesn't, and this made remembering this step a lot easier. I actually understood what was going and why the methanol treatment was so important. For future experiments, I definitely plan to learn about the theory and mechanisms behind the materials used for the experiment.