NMR Metabolomic Analysis of Biomarkers for Mouse MC3T3 Osteoblastic Cells Exposed to Ionizing Radiation

Mentors: Louis S. Kidder, Ph.D. (Radiology), Susanta Hui, Ph.D. (Therapeutic Radiology), Bruce E. Hammer, Ph.D. (Radiology)The research is relevant to radiation therapy for bone cancer. This study seeks to determine if radiation damage to bone is observable via NMR spectroscopy of cell metabolomics. Sample preparation, NMR spectroscopic techniques and analysis were developed during the previous UROP study of osteoblastcells differentiation. We hypothesize that radiation dose and time following irradiation will influence the in vitro phenotypic expression of pre osteoblasticcells. We anticipate that in vitro radiation doses 0 and 10Gy will result in a increase in the appearance of adipocytes with a related decline in osteoblasticcells. 1D-1H NMR (700MHz) spectra of water soluble extracts from MC3T3 Osteoblasticcells were obtained. The metabolomicsignature of osteoblastcells, irradiated and control , were compared through 1D-1H NMR spectroscopy . 1H NMR Spectra was analyzed by Chenomx6.1 software which is available at the University of Minnesota Biomedical NMR facility. Chenomxdeconvolvesthe chemical shifts of an NMR spectrum and assigns them to known metabolite signatures. This allows the identification of roughly 100 to 200 metabolites. The techniques developed here will be used in future experiments for cells that are magnetically levitated to observe changes in cell metabolomicswhen gravitational loading is reduced or eliminated.This project continuation is supported by the Undergraduate Research Opportunities Program (UROP) and computational resources from Minnesota Supercomputing Institute for Advanced Computational Research of the University of Minnesota