Controlling oncogenic transformation of mesenchymal stem cells by targeted epigenetic intervention

Spontaneous oncogenic transformation of mesenchymal stem cells (MSCs) is a risk factor when expanding stem cells in culture for therapeutic intervention. Transformed MSCs devide indefinitely and loose their ability to differentiate, arguing for epigenetic alterations of several classes of genes, which cannot be easily reversed. Little is known on epigenetic transitions characterizing MSC transformation, thus no approach has been taken to alleviate or reverse MSC transformation. The goal of the project is to characterize epigenetic defects associated with oncogenic transformation of human MSCs, and use this information to elaborate an epigenetic- and/or differentiation-based approach to prevent or reverse the transformed state. The system will rely on bone marrow MSCs that spontaneously transform in vitro and give to tumors. These cultures can be examined before, during and after transformation. First, we will build a map of DNA methylation, histone modification and transcriptional transitions during trans formation. Second, we will address mechanisms behind epigenetic changes associated with transformation by assessing the efficiency of gene targeting of relevant chromatin remodeling enzymes. Third, we will determine whether epigenetic defects associated w ith transformation can be prevented or reversed. The results will provide insights on how to establish safe long-term MSC culture for therapeutic goals. The project will rely on cell culture, in vitro and in vivo analyses, and epigenetic tools supported by expert bioinformatics. Collaborators in the fields of tumorigenesis, bioinformatics, DNA methylation and shRNA viral constructions will aid in this 3-year project. Personnel to be hired are two postdocs/researchers including one bioinformatician, and o ne technician versed in stem cells and DNA methylation.