Author: Devan Mishra | Major: Biochemistry | Semester: Fall 2023

During the Fall 2023 semester, I continued my honors thesis research in Dr. Greene’s Cachexia Laboratory within the Exercise Science department. Cancer cachexia is an extreme weight loss and muscle wasting syndrome that affects ~ 50-80% of all cancer patients. This syndrome cannot be reversed through nutritional support and ultimately leads to functional impairment and reduced effects of cancer treatments such as chemotherapy. The overall goal of our current study is to examine the effects of the genetic targeting of two genes (BNIP3 and OPA1) in cancer-induced mice. The specific aims of my research are to utilize our Nikon Eclipse Ti-S inverted epi-fluorescent microscope, MATLAB software, and mitochondrial respiration analysis tools to quantify and analyze different aspects of mitochondrial health, which precedes the loss of skeletal muscle mass seen in cancer cachexia. Our hope is that the targeting of BNIP3 and OPA1 will mitigate the effects of cancer cachexia, which could lead to the development of possible gene-targeted therapeutics in the field of cancer research and other chronic conditions associated with cachexia.

I first came across Dr. Greene’s work when looking for a research lab to work in during the summer after my freshman year of college. As a biochemistry major with a passion for medicine and fitness, I was drawn to the Cachexia Laboratory’s focus on the unique relationship between metabolic functions and its impact on skeletal muscle once diseases like cancer occur within the body.

With most of my honors research focusing on data analysis, a certain chunk involves fluorescent microscope imaging at various magnifications to capture crisp, clear images of the fluorescent MitoTimer protein inserted into the right flexor digitorum brevis muscle of the cancer-induced mice we harvest. Throughout my time in the Cachexia Laboratory, I have learned that a good quality image perfectly encapsulates the story of the mitochondrial health. However, this process can be tedious and difficult at times. There is a certain level of subjectivity when finding a good spot within the muscle fiber to start imaging, especially at higher magnifications such as 100x, because the sea of fibers seemingly blends together. A good trick my mentor/PhD student Fran has told me is to look for isolated fibers that have lines stretching across the fiber in both directions under the red-colored tetramethylrhodamine isothiocyanate (TRITC) fluorescent channel, which has helped immensely in capturing the impact of cancer on the mitochondrial degeneration within the muscle fibers.

Dr. Greene and my mentor Fran have both played significant roles in my development as an undergraduate researcher. From a general standpoint, I’ve gained a greater understanding of the research process and the amount of time, effort, and communication amongst the lab that goes into completing a project that yields scientific information on the forefront of medicine. More specifically, they have allowed my honors thesis research to have a certain level of flexibility, which allows me to extend beyond imaging/data analysis into other areas of focus within our lab. For example, I often participate in of our weekly harvests where I do tissue weights and focus on freezing certain tissues in liquid nitrogen. I have also had the opportunity to run the mitochondrial respiration chambers, where a certain muscle tissue from the mice is placed in a chamber simulating in-vivo conditions.

Next semester, I plan to wrap up my honors thesis by finishing the imaging for the remaining mice from our project. I will also complete the quantification of our results through data analysis with MATLAB software and other graphing tools. During this time, I also plan to learn PCR and complete the animal training protocol to become more involved with handling our mice. I also plan to present my research at a conference taking place later in the semester.