Title

Dissecting the Mechanism Behind Spy1-Mediated Expansion of Brain Tumour Initiating Cells

Submitter and Co-author information

Jake FrankFollow
Dorota Lubanska
Lisa Porter

Standing

Undergraduate

Type of Proposal

Oral Research Presentation

Challenges Theme

Building Viable, Healthy and Safe Communities

Faculty

Faculty of Science

Faculty Sponsor

Lisa Porter

Abstract/Description of Original Work

In the normal brain, tight regulation and balance between self-renewal and differentiation of neural stem cells is essential for the maintenance of homeostasis. Abrogation of that balance is characteristic for the populations of Brain Tumour Initiating Cells (BTICs), which share properties with neural stem cells, including the capacity to self-renew. Deviation from asymmetric division and differentiation results in symmetrically dividing BTICs driving progression, therapy resistance and relapse of patients with the most aggressive type of brain tumour, Glioblastoma multiforme (GBM). The cell cycle controls cellular fate decisions between self-renewal and differentiation. An atypical cell cycle regulator, Spy1, has been shown to induce acquisition of BTIC characteristics in neural stem cells in vitro and drive symmetric expansion of BTICs derived from GBM patients. The aim of this project is to determine the mechanism by which Spy1 promotes symmetry of division. We have utilized the shRNA and CRIPR/Cas9 system to deplete Spy1 in a panel of GBM patient lines to study the effects on proliferation and the mode of division. Although several lines demonstrated sensitivity to Spy1 depletion, we found lines which presented no change to proliferation rate or symmetry of division. We will utilize this panel to further study the cell cycle profile, expression and the localization of the cell cycle and checkpoint proteins and proteins implicated in symmetry of division. Immunocytochemistry, Western Blotting and qRT-PCR will be used to delineate differences between Spy1-“sensitive” and -“resistant” lines and determine key molecular players to further dissect the specific mechanism of Spy1 mediated effects in BTIC.

Start Date

31-3-2020 8:30 AM

End Date

31-3-2020 8:30 AM

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Mar 31st, 8:30 AM Mar 31st, 8:30 AM

Dissecting the Mechanism Behind Spy1-Mediated Expansion of Brain Tumour Initiating Cells

In the normal brain, tight regulation and balance between self-renewal and differentiation of neural stem cells is essential for the maintenance of homeostasis. Abrogation of that balance is characteristic for the populations of Brain Tumour Initiating Cells (BTICs), which share properties with neural stem cells, including the capacity to self-renew. Deviation from asymmetric division and differentiation results in symmetrically dividing BTICs driving progression, therapy resistance and relapse of patients with the most aggressive type of brain tumour, Glioblastoma multiforme (GBM). The cell cycle controls cellular fate decisions between self-renewal and differentiation. An atypical cell cycle regulator, Spy1, has been shown to induce acquisition of BTIC characteristics in neural stem cells in vitro and drive symmetric expansion of BTICs derived from GBM patients. The aim of this project is to determine the mechanism by which Spy1 promotes symmetry of division. We have utilized the shRNA and CRIPR/Cas9 system to deplete Spy1 in a panel of GBM patient lines to study the effects on proliferation and the mode of division. Although several lines demonstrated sensitivity to Spy1 depletion, we found lines which presented no change to proliferation rate or symmetry of division. We will utilize this panel to further study the cell cycle profile, expression and the localization of the cell cycle and checkpoint proteins and proteins implicated in symmetry of division. Immunocytochemistry, Western Blotting and qRT-PCR will be used to delineate differences between Spy1-“sensitive” and -“resistant” lines and determine key molecular players to further dissect the specific mechanism of Spy1 mediated effects in BTIC.