The evaluation of patients with rare blood disorders has uncovered fundamental aspects of biology, pathophysiology and therapies. The affected gene for several rare genetic diseases has been recently isolated, including Diamond Blackfan anemia, Shwachman Diamond syndrome, Fanconi's anemia. The derivation of animal models for these diseases has proven difficult, as the mouse mutant phenotype does approximate the human disease in many cases. Here we propose to develop human iPS induced pluripotent cell lines from patients with genetic blood diseases (including the disorders listed above and Down's Syndrome). We plan to obtain fibroblasts from patients with the diseases, and introduce Oct4, Sox2, Klf4 and c-myc as retroviruses or lentiviruses. iPS lines will be derived based on cell morphology and pluripotency marker expression. These iPS cell lines will be characterized for their ability for form teratomas. Furthermore, we plan to characterize the biology of the disease in vitro by differentiating the iPS cells to form hematopoietic cell types in colony assays as well as by transplanting the differentiated cells into immunodeficient mice. A database will be constructed to help track the cell lines, provide information on their characterization, and to aid distribution to the community. We then plan to develop a screening system to find small molecules or shRNAs that will rescue the disease phenotype in vitro. These chemicals or genes will help understand the biology of the disease, establishing the pathogenesis and ultimately finding new therapies. Furthermore, this work should pave the way for the use of iPS cells for the study of other organ systems.
Some genetic diseases of the blood system have been difficult to study because animal models have not accurately modeled the human diseases. We plan to make iPS cell lines and to try to find chemicals or genes that would rescue the diseases and ultimately prove to be new therapies.
