title: PCM1 Depletion Inhibits Glioblastoma Cell Ciliogenesis and Increases Cell Death and Sensitivity to Temozolomide.
abstract: A better understanding of the molecules implicated in the growth and survival of glioblastoma (GBM) cells and their response to temozolomide (TMZ), the standard-of-care chemotherapeutic agent, is necessary for the development of new therapies that would improve the outcome of current GBM treatments. In this study, we characterize the role of pericentriolar material 1 (PCM1), a component of centriolar satellites surrounding centrosomes, in GBM cell proliferation and sensitivity to genotoxic agents such as TMZ. We show that PCM1 is expressed around centrioles and ciliary basal bodies in patient GBM biopsies and derived cell lines and that its localization is dynamic throughout the cell cycle. To test whether PCM1 mediates GBM cell proliferation and/or response to TMZ, we used CRISPR/Cas9 genome editing to generate primary GBM cell lines depleted of PCM1. These PCM1-depleted cells displayed reduced AZI1 satellite protein localization and significantly decreased proliferation, which was attributable to increased apoptotic cell death. Furthermore, PCM1-depleted lines were more sensitive to TMZ toxicity than control lines. The increase in TMZ sensitivity may be partly due to the reduced ability of PCM1-depleted cells to form primary cilia, as depletion of KIF3A also ablated GBM cells' ciliogenesis and increased their sensitivity to TMZ while preserving PCM1 localization. In addition, the co-depletion of KIF3A and PCM1 did not have any additive effect on TMZ sensitivity. Together, our data suggest that PCM1 plays multiple roles in GBM pathogenesis and that associated pathways could be targeted to augment current or future anti-GBM therapies.
authors.affiliation: Department of Neuroscience, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA; Preston A. Wells, Jr. Center for Brain Tumor Therapy, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA.
authors.forename: Lan B
authors.initials: LB
authors.lastname: Hoang-Minh
authors.affiliation: Preston A. Wells, Jr. Center for Brain Tumor Therapy, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA; Department of Neurosurgery, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA.
authors.forename: Loic P
authors.initials: LP
authors.lastname: Deleyrolle
authors.affiliation: Department of Neuroscience, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA.
authors.forename: Nariaki S
authors.initials: NS
authors.lastname: Nakamura
authors.affiliation: Department of Neuroscience, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA.
authors.forename: Alexander K
authors.initials: AK
authors.lastname: Parker
authors.affiliation: Preston A. Wells, Jr. Center for Brain Tumor Therapy, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA; Department of Neurosurgery, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA.
authors.forename: Regina T
authors.initials: RT
authors.lastname: Martuscello
authors.affiliation: Preston A. Wells, Jr. Center for Brain Tumor Therapy, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA; Department of Neurosurgery, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA.
authors.forename: Brent A
authors.initials: BA
authors.lastname: Reynolds
authors.affiliation: Department of Neuroscience, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA; Preston A. Wells, Jr. Center for Brain Tumor Therapy, University of Florida College of Medicine, McKnight Brain Institute, Gainesville, FL 32610, USA. Electronic address: msarkisian@ufl.edu.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Sarkisian