Erin Swanson, a sophomore studying biochemistry at St. Norbert College, evaluated the stability and localization of XPR1:KIDINS220 through intracellular flow.
Precision cancer medicine promises to understand the specific molecular events in cancer cells, thereby identifying unique therapeutic strategies to kill cancer while sparing normal tissues. The Broad is an incredible place filled with incredible scientists who, together, encompass a true passion for scientific discovery. Learning and growing alongside this community has been an absolute privilege as I continue cultivating and exploring my passion for science. In ovarian cancer, high levels of the phosphate importer SLC34A2 have been linked to a cellular dependency on the phosphate export complex XPR1:KIDINS220. Both proteins are required for cellular phosphate efflux, but little is known about the mechanistic underpinnings of phosphate efflux. To develop a scalable method for evaluating the stability and localization of XPR1:KIDINS220, we turned to flow cytometry. We are developing methods for analyzing the total cell surface expression of the complex as well as total protein levels using an intracellular staining protocol. Initial staining results have shown successful antibodies binding directly to XPR1 but not KIDINS220. As an alternative approach, PRIME genome editing will be utilized to tag KIDINS220 with epitope tags endogenously. With an established flow cytometry protocol in place, we will then apply functional genomics tools like CRISPR base editing or CRISPR knockout screens to identify regions across XPR1:KIDINS220 that are required for proper localization and stability. Systematically identifying the mechanisms that govern localization and stability will give us a better understanding of the mechanisms of phosphate efflux. We hope these studies will nominate novel strategies for inhibiting XPR1:KIDINS220 for treating ovarian cancer.
Project: Evaluating the stability and localization of XPR1:KIDINS220 through intracellular flow
Mentor: Daniel Bondeson
PI: Golub Lab, Cancer Program