Interdisciplinary Training

Sara Fagerlie

"Generation of Fanconi Anemia Group B Canine Embryonic Stem Cells"

Fanconi Anemia (FA) is a heritable chromosome instability syndrome characterized by cellular hypersensitivity to DNA cross-linking agents, multiple congenital abnormalities, progressive bone marrow failure and a high incidence of malignancy. Ten autosomal recessive and one X-linked FA disease-causing genes have been cloned and characterized. However, despite progress made in defining the role of the FA proteins with respect to DNA repair, the pathogenesis of certain FA features, including bone marrow failure, the most lethal and ubiquitous FA phenotype, remains unclear. Moreover, the only existing FA model, the mouse, does not mimic FA patient hematopoietic defects. Therefore, we propose to target the X-linked Fancb gene in canine embryonic stem (cES) cells to generate a pluripotent FA group B (FA-B) embryonic stem cell line. Achieving these goals will generate a novel model system that can be used to determine proteomic changes induced by the FA-B phenotype before and after differentiation. Once methodologies are established to drive hematopoietic differentiation, hematopoietic differentiated cES will then be transplanted into a myeloablated normal dog and used to study FA hematopoietic cells in the context of a normal microenvironment. Examining function in the absence of confounding FA auxiliary cells will be a valuable way to isolate aberrant functions of the FA hematopoietic system and defects specific to hematopoietic cell lineages, aiding in further development of the only known FA treatment-FA specific stem cell transplant.