Implication of Orphan Nuclear Receptors Nr4a1 and Nr4a3 in a Conditional Knockout Murine Model for Acute Myeloid Leukemia
Open Access
- Author:
- Harrington, Emily Ann
- Area of Honors:
- Biology (Behrend)
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Michael A Campbell, Thesis Supervisor
Michael A Campbell, Thesis Honors Advisor
James Warren Jr., Faculty Reader - Keywords:
- acute myeloid leukemia (AML)
hematopoiesis - Abstract:
- Limited understanding of the mechanisms that control the proliferation of cancerous cells such as those found in acute myeloid leukemia (AML) is among one of the most important reasons why we do not know how to treat cancers appropriately. Further investigation into the way that cancers develop is our current hope in developing new treatments for disease. We show in this set of experiments that genes NR4A1 and NR4A3 are implicated in the onset of AML as shown previously5. A Tamoxifen induced conditional knockout murine model with floxed NR4A1 regions was used to show that the down regulation of both NR4A1 and NR4A3 leads to AML. Hematoxylin and Eosin staining was performed upon lung, liver, and spleen samples in order to view the myeloid infiltrates that are present in the conditional knockout as opposed to wild type tissues. Bone marrow was isolated from AML mice and transplanted into lethally irradiated mice. After transplantation, these mice displayed complete engraftment of the bone marrow cells and confirmed that this disease is transplantable. In addition, an NR4A3 rescue was attempted by isolating other samples of bone marrow from the AML mice and employing retroviral infection techniques. Using Fluorescence-activated cell sorting (FACS), we isolated upper and lower side population cells from wild type and conditional knockouts in order to perform a stem cell assay. The conditional knockout stem cell colony formations displayed a skewing of lineage potential for upper-primed cells. With these data, we detail novel findings in the shift of lymphoid-primed cells to a myeloid-like phenotype, suggesting that more research will be necessary to understand this breakdown in hematopoiesis.