ICOC Funds Committed:
A major challenge in the development of regenerative medicine strategies involving human embryonic stem cells (hESCs) remains the routine and reproducible culturing of hESCs in the laboratory without loss of their developmental potential. Outside their natural environment, cell culture conditions in the laboratory represent artificial cellular environments that cause cellular stress and may harm the cell physiology. Most human cells respond to cellular stress conditions by activating mechanisms that are capable of reducing the damage to the cell. However, activating these cellular stress response mechanisms may often also have consequences for the proliferation and developmental capacity of the cells. To response DNA damage, most cells for example inhibit the progression of the cell cycle until the DNA has been repaired. In previous work, our laboratory has characterized the regulatory proteins and mechanism that control stress responses in mammalian immune cells. We are able use computational simulations of these complex regulatory mechanisms to gain insights and direct specific experiments. Here we propose to determine how stress responses are regulated in hESCs. We will make quantitative measurements regarding the presence of the key regulatory proteins and construct a computational model of the regulatory networks as it pertains to hESCs. This allows us to compare different hES cell lines, provide important insights on their physiological regulation, and predict the effects of pharmacological treatments. As the very first molecular and computational description of important regulatory mechanisms, the results will also form the basis for future studies that monitor the molecular changes during tissue development and the role these molecular components play in such tissue development. In addition, we propose to engineer molecular tools that allow us to monitor the molecular stress level in the cell. Engineered proteins inserted into hESCs will be used as sensors of cellular stress. By monitoring their activity we will optimize laboratory methods and techniques related to growing stem cells in the laboratory. These studies may have result in improving current technologies and make a critical step in developing regenerative medicine more reliable and routine.
Statement of Benefit to California:
A major challenge in the development of regenerative medicine strategies involving human embryonic stem cells (hESCs) remains the routine and reproducible culturing of hESCs in the laboratory without loss of their developmental potential. Outside their natural environment, cell culture conditions in the laboratory represent artificial cellular environments that cause cellular stress and may harm the cell physiology. The proposed research is focused on characterizing the molecular stress responses in hESCs. The research will significantly contribute to an understanding of stem cell biology and the molecular mechanism that regulate the capacity for self-renewal and for tissue development. The molecular characterization will contribute to an understanding how different hESC lines differ and what their respective advantages may be. And finally the proposed research will result in improved laboratory methodologies for the handling of hESCs. The research will benefit the State of California and its citizens by (1) enhancing the scientific knowledge base of the molecular mechanisms that control hESC physiology and developmental capacity (2) improving the methodologies for culturing and manipulating hESCs thereby increasing the likelihood of successful of regenerative medicine strategies (3) making California a leader in stem cell systems biology by constructing a computational model that allows for virtual cell studies by simulations (4) training a junior PI and two talented postdoctoral fellows with proven scientific track records in stem cell biology, enabling them to contribute to Regenerative Medicine in their own future laboratories.