Targeting the proteosome in atypical teratoid/rhabdoid tumors
Mentor Name: Eric Raabe
This project will investigate novel therapeutics to treat atypical teratoid/rhabdoid tumor (AT/RT), a highly malignant pediatric brain tumor of the central nervous system (CNS). The primary genetic abnormality of AT/RT is defined by the complete loss of function of the SMARCB1 gene, and genes in various signaling pathways are dysregulated in AT/RT cell ines--including some in the NF-kB pathway. We hypothesize that targeting the NF-kB pathway in AT/RT will prevent tumor growth and/or promote cancer cell death. To test this hypothesis, we will use two drugs--ixazomib and curaxin (CBL0137)--in combination. CBL0137 suppresses tumor growth in both AT/RT cells and medulloblastoma mouse models, while Ixazomib is a clinically approved drug for treating multiple myeloma via inhibition of the NF-kB pathway. To track changes in NF-kB activity following drug treatment in AT/RT cell-lines, we will transduce AT/RT cells with a commercially available NF-kB nano-Luc reporter construct. We will compare DMSO control, CBL0137, ixazomib, and combination therapy as well as assess luciferase expression, cell viability (CellTiterGlo), and apoptosis (cPARP western blot and CC3 immunofluorescence). We will will assess expression levels of NF-kB specific target genes, such as cyclin D1, BCL-XL, cyclin D2, and NFKBIA using qPCR, Western blotting, and immunofluorescence staining.
