Abby
Cortez
Malaria is caused by Plasmodium parasites which are deposited into the host by a mosquito bite. The parasite first travels to and infects liver cells before egressing to the bloodstream. During the liver stage, the Plasmodium parasite resides in the parasitophorous vacuole (PV). The PV membrane increases in size to accommodate the replicating parasite organelles, membranes, and nuclei. For these events to transpire, the parasite needs an abundance of nutrients including sphingolipids. Previous research has demonstrated that sphingolipids are upregulated in Plasmodium berghei infected HuH7 cells and that they are traPicked to the Plasmodium parasite to aid in their development. Ceramide transport protein (CERT) could potentially play a role in sphingolipid traPicking to the PV. To study the role of CERT during the Plasmodium liver stage, we first generated a CERT CRISPR knockout cell line. We then compared parasite development between wildtype and CERT knockout cells by measuring parasite load with a plate reader assay. Immunofluorescent microscopy was also used to identify the infection rate, parasite size, and CERT localization. Lastly, the number of sphingolipids or ceramides traPicked to the PV were compared between wildtype and knockout cells to determine if CERT supports sphingolipid acquisition. Investigating the RIOK2-NFkB Interaction in Prostate Cancer
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Abby Cortez
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Right Open Reading Frame Kinase (RIOK2) is critical in ribosome assembly, which is needed for proliferating cancer cells. Targeting RIOK2 is shown to decrease tumor growth, likely due to impairment of ribosome biogenesis. Recently, our group discovered that RIOK2 has novel DNA binding activity, but there are still unknowns such as the cofactors it binds. A published high-throughput mass spectroscopy study suggests that RIOK2 interacts with RELA, a subunit of Nuclear Factor Kappa-light-chain-enhancer (NF-kB). NF- kB is a transcription factor that regulates genes essential for cancer cells. We hypothesize the RIOK2-NF-kB interaction supports disease progression. A nuclear fractionation experiment showed RIOK2 in the nucleus and cytoplasm. A co-immunoprecipitation assay of 22RV1 and PC3 cell lysates showed that NF-kB was bound to RIOK2 when not part of a ribosome complex. A drug combination assay indicated an additive ePect on proliferation when co-targeting both proteins. The results of preliminary studies validated the presence of an NF-kB -RIOK2 interaction in prostate cancer cells. Next, we will conduct a nuclear- fractionation assay targeting RIOK2 or NF-kB to understand where the interaction occurs and its regulation. Conversely, we will use NF-kB activating ligands to determine how activation of NF-kB aPects RIOK2-NF-kB binding. An understanding of this interaction is important to understand basic cancer cell biology and for potential translational interventions.
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Duke University / 2024
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Abby Cortez