Research Department: Internal Medicine - Cardiology | Molecular Biology
Graduation Date: May 2021
Abstract: The localization of intrinsically disordered regions (IDR) of liquid-liquid phase droplets,otherwise known as nuclear granules, through the use of a biotinylated compound: 𝛽-Isoxazole (𝛽-Isox). 𝛽-Isox is known to precipitate proteins with IDRs, but the focus of my project is targeting IDRs of superenhancers (SE) with 𝛽-Isox; SE’s are a cluster of enhancers, along with transcription factors, that drive high levels of transcription. Once 𝛽-Isox is localized in the nucleus, several approaches can supplement its specific target site. This includes using different cell lines such as Bj5-ta or U2OS, isolating nuclei from in vitro, introducing Digitonin or Pluronic permeabilization agents for 𝛽-Isox, concentration gradient for𝛽-Isox,and attempting different incubation time points. In addition to this, the colocalization of 𝛽-Isox is implemented through the use of different stains at the histone level for active enhancers (H3K27ac) or active promoters (H3K27me3). In conjunction to this colocalization, another approach is being applied where the transfection of cofactor vectors BRD4 and MED1, that have shown to form part of liquid-liquid phase droplets, can also be colocalized with 𝛽-Isox. Ultimately, the profiling of these IDRs, through the use of 𝛽-Isox, can reveal a relationship between nuclear condensates and gene regulation not only for our cell lines, but for human heart SA node, AV node, and other ventricular conduction system nodes.
What Does Research Mean To You? Three years ago, my first response would have been that it means being scared. Too scared to take on a new challenge and think I wasn’t cut out to be in a research lab. Just the sound of it was daunting, but I came to terms that the only reason I never wanted to try out research was because I was afraid not to be good at something I’ve never been exposed to. Today, it means exactly that: taking one step at a time, but realizing that starting was the hardest part. Take on the new challenge, be insightful, come with tons of questions, but never leave with less understanding than what you brought to the table.
Tell Us About Your Journey What really spiked my interest in research was finding out Organic Chemistry was fun! I know, I know, but really doing all those synthesis questions was like a puzzle. In the Spring of 2019, I sought out to apply the basis of chemistry and joined Dr. Jung-Mo Ahn’s biochemistry lab that emphasizes on prostate cancer and diabetes research. Heart disease and its impact has resonated within my family and is a significant motivator to pursue a biomedical career. Thus, that same summer I was blessed that the National Heart, Lung, and Blood Institute gave me the opportunity to immerse myself in cardiovascular research at Case Western Reserve University School of Medicine. That summer really gave me an insight into what the world of molecular biology was like and only encouraged me to take advantage of what the Green Fellowship had to offer so that I may find my fit as a prospective M.D./Ph.D.
How Did the Pandemic Affect Me? It was very challenging going from hands-on to strictly all online. I did most of my experiments based on a schedule with my lab manager since I didn’t have proper training to expensive equipment (confocal imaging and such). Once the news hit us I was very overwhelmed with how rapidly things developed for which even Dr. Ravnik did not know the answers to. An advantage was no more commuting and our research symposium turned out better than I expected! Take care of yourselves and others during these stressing times and even when the pandemic is over.
Where am I now? Current Academic Status: Pursuing a Master of Science in Medical Science at UNTHSC. Planned Professional Path: To pursue an MD to become a cardiothoracic surgeon with research emphasis in cardiomyopathies.
Advice for Future Green Fellows
To optimize what the Green Fellowship has to offer, make sure you come with an open mind and heart. Research is not just about the success rate or the discoveries that matter. It is the process and failures that you experience that are enriching because that is an opportunity for self-growth. That is something I am grateful the Munshi Lab over at UTSW taught me which brings up my next point. The best experience will be finding the right mentor, more so than the research itself. I was grateful to have been paired with Dr. Munshi and his team who were all very approachable and super-friendly! Research or no research experience, you’ll never know unless you apply and put forth your best effort.