A Pooled CRISPR/Cas9 screen to identify essential kinases for GBM stem cells.
Zhang, Yahan.
2019
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Glioblastoma
multiforme (GBM) is the most common primary malignant brain tumor. GBM has a very poor
prognosis, with a less than 10% 5-year survival rate. It is a WHO grade IV glioma which
are significantly more proliferative, invasive, and angiogenic than lower grade gliomas.
However, currently there is still no significant targeted therapeutic that is effective
for GBM therapy. A subpopulation ... read moreof cells enriched in hypoxic niches known as GBM
stem-like cells (GBM-SC) play an important role in initiating and maintaining tumor
growth, and are thought to drive GBM recurrence. Therefore, it is important to identify
genetic determinants for the proliferation and survival of GBM stem cells. In previous
pooled RNAi and kinase inhibitor library screens, serum and glucocorticoid-regulated
kinase 1 (SGK1) and other kinases in the RTK signaling pathway have been shown to be
essential for GBM-SC growth and survival. However, both RNAi and small molecular screens
have significant off target effects. In this study, to try to circumvent this problem, I
have worked to develop a pooled CRISPR kinome screen targeting over 500 kinases to more
definitively identify key kinases for GSCs' survival and proliferation. I have shown
that I can infect and identify cells that have been infected with lentiviruses targeting
the entire human kinome. Unfortunately, the quantitative analysis of my data at this
time, has not allowed for a definitive identification of all essential kinases. Here I
discuss these results and suggest methods to obtain more definitive CRISPR screening
results. Hopefully the data from this study will help to lead to the identification of
new therapeutic targets for GBM.
Thesis (M.S.)--Tufts University, 2019.
Submitted to the Dept. of Pharmacology and Drug Development.
Advisor: Brent Cochran.
Keyword: Pharmacology.read less - ID:
- dv1406842
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