Pavankumar Umashankar
MIT Department: Mechanical Engineering
Faculty Mentor: Prof. Ritu Raman
Research Supervisor: Tamara Rossy
Undergraduate Institution: University of Maryland, Baltimore County
Hometown: Ellicott City, Maryland
Website: LinkedIn
Biography
Pavan Umashankar is a double-major undergraduate in Biochemical Engineering and Biochemistry at the University of Maryland, Baltimore County (UMBC). He has a passion for medicine, biomedical research, and service and plans to pursue a dual-degree M.D./Ph.D. program after graduation. Pavan works with Prof. Ritu Raman and Dr. Tamara Rossy at MIT to better characterize muscle fibers through myosin immunostaining to help better understand neurodegenerative diseases where muscle fiber types play central roles. His research at UMBC involves understanding the cell and molecular dynamics of proteins implicated in ovarian cancer pathology. He is passionate about working on similar projects that involve a better understanding of cell/tissue/system characteristics (with a special interest in cancer biology and aging science) and using engineering perspectives to design better therapeutics during his M.D./Ph.D. education. Pavan is also passionate about working alongside individuals with healthcare needs, industry partners, and other community stakeholders to more efficiently implement biomedical innovations.
Abstract
Multi-pronged Skeletal Muscle Fiber Type Characterization through
Immunostaining, Contraction Analysis, and qPCR
Pavankumar Umashankar1, Tamara Rossy2, Ritu Raman2
1Department of Chemical, Biochemical and Environmental Engineering, University of Maryland Baltimore County
2Department of Mechanical Engineering, Massachusetts Institute of Technology
Skeletal muscles mediate all voluntary movement and are composed of different fiber types that enable a wide range of functionality by tuning contractile strength and metabolic needs. Muscle fiber types change with physiological and pathophysiological stimuli. To study fiber type transformation in vitro, we need a way to reliably characterize and monitor fiber types. In this study, we optimized a three-pronged strategy to map fiber types in engineered muscle tissues differentiated from murine C2C12 myoblasts: immunostaining, quantitative PCR (qPCR), and analysis of contraction dynamics in response to electrical stimulation. These studies enabled us to understand how varying fiber morphology and gene expression impacts function.
After an initial screening to identify optimal dilutions and combinations of antibodies
against specific myosin heavy chain isoforms representative of slow- and fast-twitch fibers, we
investigated the changes in fiber type composition throughout the muscle maturation process. We analyzed muscle fibers on day 6, 10, and 14 of differentiation through stimulated contraction and calcium imaging assays, and performed qPCR and immunostaining analysis at the end of the experiment. Overall, the optimization of this three-pronged strategy has enabled better investigation of fiber type changes, with applications ranging from medicine to biohybrid machines.
