Generating Reprogrammed Pluripotent Human Cell Lines Using a Histone Modification Barcode
Human embryonic stem cells (hESCs) hold significant promise for the future of cell-based therapeutics through their ability to differentiate into any cell lineage. A major consideration for cell transplantation therapy is to transplant cells that will not be rejected. The most effective way to do this will involve deriving hESC lines from a patients own cells. This is achieved through dedifferentiation of an adult cell back to a pluripotent state in a process known as reprogramming. For successful reprogramming three fundamental problems need to be tackled: 1. Identify an efficient reprogramming strategy; 2. Develop a comprehensive in vitro assay to measure reprogramming success; and 3. Understand the mechanism by which reprogramming occurs in order to prevent reprogramming towards cancerous transformation. In the current proposal we aim to develop a comprehensive in vitro assay to measure reprogramming success, and use this assay to monitor reprogramming of an adult human cell to a pluripotent stem cell. Results from this proposal will generate a sensitive and specific reprogramming diagnostic tool that measures genome-wide nuclear changes that associate with formation of a reprogrammed pluripotent cell type. This will significantly improve success rates of reprogramming, and enhance the methods for generating therapeutically useful cell types.
Human embryonic stem cells (hESCs) hold significant promise for cell-based therapeutics for diseases including HIV/AIDS, Alzheimer’s and neurodegenerative disorders such as Parkinson’s and MS/ALS, type I diabetes, heart attack, stroke, and tissue repair following devastating injury such as damage to the spinal cord. For treatment using cell-based therapies, the derivation of individualized hESC lines would be ideal, since genetic differences between individuals prevents a `one cell line fits all’ approach. The only way to generate individualized hESC lines is through the process of nuclear reprogramming. At the current time, nuclear reprogramming of human cells is extremely inefficient. In this research proposal, we aim to improve the existing methods for monitoring nuclear reprogramming. We also aim to use this information to reprogram human cells without the requirement of a woman’s oocyte for somatic cell nuclear transfer. Research findings from this proposal will provide a much-needed barometer for reprogramming success allowing the generation of individualized human stem cells.