Using CRISPR to Explain the ART of Artemisinin Analogs
Suhayl Khan
Senior, Health Science
Benedictine University
2019 STRIDE Fellow

My Research Project

Diagram showing how the CRISPR-Cas9 editing tool works.

Artemisinin is a drug derived from the Artemisia annua plant. It is known for its anti-malarial properties, but has also been found to have anti-cancer properties. The active portion of artemisinin is an oxygen-oxygen bond called an endoperoxide. When in contact with free iron in a cell, this endoperoxide breaks and creates oxygen radicals which are extremely reactive. These oxygen radicals then proceed to react with cellular components such as membranes and proteins which eventually leads to cell death. Previously, it had been found that DMR1 and HSM2—two analogs of artemisinin— are particularly effective in inducing cell death in cancer cell lines but not in normal cell lines. This summer, my lab and I worked on figuring out why this is so.

 

It has been found that cancer cells contain a higher iron concentration than normal cells. This higher iron concentration is due to higher concentrations of transferrin receptors—the receptor that transports iron into the cell— in cancer cells when compared to normal cells. We believe that the specificity of our artemisinin analogs to cancer cells is due to the higher concentration of iron in cancer cells. To test this, we planned to use Clustered Regularly Interspaced Short Palindromic Repeats, or CRISPR, a gene editing technique that can remove the transferrin receptor gene in lung cancer cells. Then, we would test our analogs on these transfected cells to determine if a lower iron concentration would show the analogs as ineffective. However, we were unable to test our analogs on the transfected cells because the transfected cells died three days after successful transfection. This proved to us that transferrin receptor is required for cell growth and the proliferation of cancer cells, and that cancer cells cannot survive with low iron concentrations. In the future, we plan on using CRISPR to overexpress the transferrin receptor gene in normal lung cells and testing our analogs on these cells to see if the specificity of our artemisinin analogs is indeed due to iron concentration within the cell.

Realities of Research

Cell culture flasks and media in a laminar airflow hood.

Doing research this summer has been very enlightening. In all honesty, before starting research, I imagined it to be a bit boring. I couldn’t see myself really enjoying sitting at a bench and waiting for experiments to run and cells to grow. Surprisingly, when doing research on a subject that you enjoy, it all becomes very exciting. I have learned so much about cell culture techniques and how to maintain a lab this summer. I found myself waiting in anticipation for an experiment to finish because I was so curious to know the results.  I couldn’t wait for cells to grow to large, usable percentages because I wanted to get the next experiment running. Admittedly, it was always disappointing when certain experiments didn’t go as planned or when a lengthy experiment needed to be done multiple times due to errors in previous runs. However, I have learned that even when experiments yield unexpected results, those results still contribute to the research we are conducting. It is not uncommon for an experiment to produce strange results that only make sense after hours of thinking “How could this have happened?” Fortunately, all data that we obtained this summer—expected and unexpected—contributed to my original hypothesis

Life as a Scientist

My day-to-day life as a scientist consisted of waking up early, getting to lab and checking on the cells. Every Monday, Wednesday and Friday the cells have to be fed. If they have grown exponentially, they needed to be split into a new flask. The cell media must be warm, so I had to turn the water bath on and place tubes of media in the bath well before I needed them. I checked the cells under a microscope and estimated the amount of cell growth of each individual flask. If a flask had less than 80% cell growth, the media needed to be discarded and replaced. If a flask had cell growth of 80% or above, then the cells needed to be removed from the current flask and placed into a new one to give them more room to grow. After feeding and splitting was completed, I met with my research mentor and discussed what needed to be done for the rest of the day. The biggest surprise about being a scientist was realizing how little I know about my field of research. Going into research, I believed that I had decent knowledge of physiology and biochemistry. Despite this, I spent every day learning something new and interesting about these fields. My favorite part about research is that there always seems to be more to do. Because of this, there was never a moment where I was bored with nothing to do. That being said, my least favorite part was that there were certain days where an experiment was particularly long and I found myself either overwhelmed with the amount of work to be done or exhausted by the amount of work I completed. Fortunately, working as part of a lab team took a huge amount of stress and burden off of my shoulders. It was very nice to have people to talk to and help me out whenever I need help with a task. Overall, life as a scientist is very rewarding and I have learned so much since I started research this summer.

Suhayl Khan is a senior majoring in health science at Benedictine University in Lisle, Ill. He is a 2019 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Jayashree Sarathy’s lab at Benedictine University. Suhayl’s fellowship is funded by the American Physiological Society and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduation, Suhayl plans to pursue a Master of Healthcare Administration or Master of Public Health.

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