Session 3G: Uncovering the Role of Tumor-Associated Antigen IL13Ra2 in Glioblastoma Progression
Session Number
Session 3G: 1st Presentation
Advisor(s)
Drs. Irina V. Balyasnikova and Michael Chastkofsky, Northwestern University
Location
Room A115
Start Date
26-4-2018 12:40 PM
End Date
26-4-2018 1:25 PM
Abstract
Glioblastoma (GBM) is the most aggressive brain tumor with no cure. GBM is characterized by its invasiveness, rapid growth, and therapeutic resistance. IL13Rα2 receptor is overexpressed in GBM, and its high expression inversely correlates with patient survival. However, the functional role of IL13Rα2 is not known. Our data demonstrated that the partial knockdown of IL13Rα2 with two shRNAs alters the expression of a network of transcription factors. Analysis of a human protein atlas revealed the nuclear and transmembrane localization of IL13Rα2 in glioma cells. We, therefore, focused on the validation of subcellular localization of IL13Rα2 in glioma cells. We utilized PCR, Western blotting, fluorescent and confocal microscopy, cell culture, transfection with plasmid DNA, and cell fractionation to answer this question. Nuclear localization of IL13Rα2 was observed in several glioma cell lines using microscopy. However, cell fractionation and staining of the nuclear fraction for this protein did not confirm the result. Furthermore, analysis of 293T cells transfected with IL13Rα2 tagged with myc protein revealed cytoplasmic, but not nuclear, localization of IL13Rα2. Currently, we are utilizing CRISPR to knock out IL13Rα2 in order to confirm our results obtained with shRNAs and better understand its functional role in GBM.
Session 3G: Uncovering the Role of Tumor-Associated Antigen IL13Ra2 in Glioblastoma Progression
Room A115
Glioblastoma (GBM) is the most aggressive brain tumor with no cure. GBM is characterized by its invasiveness, rapid growth, and therapeutic resistance. IL13Rα2 receptor is overexpressed in GBM, and its high expression inversely correlates with patient survival. However, the functional role of IL13Rα2 is not known. Our data demonstrated that the partial knockdown of IL13Rα2 with two shRNAs alters the expression of a network of transcription factors. Analysis of a human protein atlas revealed the nuclear and transmembrane localization of IL13Rα2 in glioma cells. We, therefore, focused on the validation of subcellular localization of IL13Rα2 in glioma cells. We utilized PCR, Western blotting, fluorescent and confocal microscopy, cell culture, transfection with plasmid DNA, and cell fractionation to answer this question. Nuclear localization of IL13Rα2 was observed in several glioma cell lines using microscopy. However, cell fractionation and staining of the nuclear fraction for this protein did not confirm the result. Furthermore, analysis of 293T cells transfected with IL13Rα2 tagged with myc protein revealed cytoplasmic, but not nuclear, localization of IL13Rα2. Currently, we are utilizing CRISPR to knock out IL13Rα2 in order to confirm our results obtained with shRNAs and better understand its functional role in GBM.