Cancer research and scoring?
02 July 2021
Hi! My name is Darline Murat and I’m a recent graduate from the University of Delaware (UD). I am the proud bearer of a Bachelors of Arts with Distinction in the Biological Sciences with minors in Genetic Counseling, and Dance.
The road to the UD-FCCC research fellowship began in my junior year, during my time as a McNair Program Scholar. During that experience, I studied topoisomerase II’s function, (an enzyme that relieves DNA supercoiling by cutting both DNA strands simultaneously and sealing them) during embryogenesis in a genetics laboratory at UD. Upon completing the program, I found that I actually enjoyed performing research, so much so that I searched for more research opportunities to broaden my knowledge and experience. One particular area of research that caught my attention was cancer research. The next semester I mentioned this new interest to my advisor--Carly Meluney--who in turn told me about an opportunity at Fox Chase Cancer Center: a research fellowship for UD students interested in exploring their interests in cancer research--what could be more perfect? After taking a tour of the cancer center, I immediately became interested in the site. The staff seemed friendly and humble and the researchers I met were so enthusiastic about their work. Not only that, but the measures they took to make the facility more welcoming to their patients, such as the paintings and sculptures displayed in the Women’s Cancer Center, sold it for me and I knew I had to research here during the 2020 summer. After applying to the research fellowship, I was glad to be one of the chosen four to complete research that summer, however, COVID-19 had other plans.
Fast forward another year to Summer 2021 and I am finally here (at Fox Chase) and am loving it! During my first days in the lab, I met everyone in my lab--Dr. John Whetstine’s epigenetics group--and even shadowed my labmates as they performed fluorescence in-situ hybridization (FISH) experiments.
For FISH, the nuclei are placed on glass slides, and then stained with DAPI (to label DNA) and fluorescent probes that bind to specific gene regions of interest. These slides are then imaged and later scored. Scoring, counting the number of probes (ie. the copy gains) within each nuclei; seems simple enough, right? WRONG! Lord help me, I’m finding this to be more difficult than anticipated but mark my words, I WILL have at least one accurate score by the end of this internship. Everything in the lab is fast-paced and I am more than up to the challenge. I am working alongside three amazing researchers: Reuben, Zach, and Gulnaz. The lab investigates lysine demethylases (KDMs) and how KDM inhibition or overexpression causes copy gains in specific chromosome regions that are often found in cancer cells. Yes, that. These copy gains are troublesome within cancer cells because they often occur within gene regions that promote cell division. These copy gains contribute to uncontrolled cell proliferation (aka tumors). Not only that, but these copy gains are ironically associated with the use of chemotherapy drugs which may contribute to a cell’s overall resistance to a drug.
In regards to the other methods used in the lab, I think I’ve finally gotten the hang of cell splitting, FISH slide preparation, and am making improvements on quantitative PCR….slowly but surely. With my gradual progress in lab techniques it’s sort of bizarre knowing I’m half way through the fellowship; there’s so many techniques and methods I want to practice. Fortunately, I am practicing protein quantification, Western blots, soon. I can’t wait for my first Western with Gulnaz next week! Even if the membrane comes out wrong, I’m sure I’ll still have better success with that than scoring. Wish me luck!
Research Kills!
29 July 2021
These next few weeks were nothing short of eventful. The three members of my cohort and I were finally able to schedule some time to hang out together. We went mini golfing and on my third attempt at hitting the ball (after missing the ball completely) I got a HOLE IN ONE on the first shot! Not too shabby considering that I never touched or hit a golf ball prior to this outing. I also went to an escape room for the first time. If I am being honest, it was quite satisfying making it out of the room with 30 seconds to spare, especially since the room was the second hardest room on the site. I’ll probably try more escape rooms in the future.
I also met a former NFL player! Of all the people Dr. John Whetstine could know: how is he friends with Chris Draft? It amazes me how these two men from completely unrelated occupations are able to come together and promote the White Ribbon Project. The White Ribbon Project was created to promote lung cancer awareness which I personally think is great since everyone has a chance of developing lung cancer whether they have a smoking history or not. Can’t wait for the ribbon making event next month. Hopefully by the end of this internship my PI’s networking skills rub off on me too!
You might be wondering what I’ve been doing at Fox Chase since my last blog entry, and whether my quantitative PCR (qPCR) skills have improved? YES! I am officially referring to myself as the qPCR Queen. The last eight or ten qPCRs came out beautifully so I think it's safe to say that I have mastered the art of micropipetting (hears applause in the background…). In respect to my first western blot, she came out GORGEOUS. I am now two for three on successful Western blotting. My scoring accuracy has also improved and I was able to help Gulnaz, one of my research mentors, with a new technique, immunofluorescence. Immunofluorescence (IF) uses antibodies and antigens to target the cell or protein of interest; my lab uses it to image histone modifications. One of the buffers used for IF uses sodium azide which is highly toxic to the body when touched or inhaled. Luckily the quantities used in the buffer are minute so it’s not lethal. I find it ironic that in order to conduct research, scientists are always putting themselves in harm's way when working with hazardous chemicals. It definitely gives a new perspective to research.
Okay, last but not least, I was able to shadow in the clinical genetics office this month. It was great observing patient appointments and interviewing a few counselors about what their jobs look like when they are not in appointments. If you have horrible listening skills this field is not for you. The fact that these counselors were able to remember the key details during their appointments is astounding. So thankful for this opportunity. See you in my next entry!
This one’s short. I promise!
12 August 2021
My final entry. These last few weeks of the fellowship went by rapidly. My lab took me mini golfing to celebrate the end of the fellowship, the new experiences and growing friendships. Unfortunately, I did not get a hole-in-one and I kept sending my ball into the bushes (maybe I should’ve tried actual golfing instead of mini golfing).
During the fellowship, I practiced a variety of lab techniques including some that I had never tried before like transfection (a method used to insert the small interfering RNA into the cell), RNA extraction, and cDNA synthesis. I am happy to write that I am officially three for four on successful Western blotting. Go me!
Beyond the lab, I had the chance to present my summer project at the virtual symposium….after editing my presentation a thousand times giving (what felt like) untold numbers of practice talks. Looking back at the summer and all of the rookie mistakes that I made now don’t seem so grave (plus my mentor got a freshly cleaned fume hood out of my most traumatic mistake--bonus!). All in all I think I consider this summer successful and I thank everyone who has helped me learn, explore, and grow.
