Thesis

Investigation of Gene Dependencies in Paediatric Tumour Models Using CRISPR-Cas Screening Technology in Providing Novel Therapeutic Targets for Paediatric Cancer

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Paediatric cancer accounts for the majority of cancer-related mortality in children. Although current
treatment increased the overall survival rates, young individuals often encounter life-long side effects
from toxic therapies. Therefore, there is an urgent need for targeted therapies aimed at
tumour-specific vulnerabilities for greater specificity and reduced toxicity, in which precision medicine
(e.g., pooled CRISPR screening and drug screening) comes into play. The objective of this research is
to identify novel gene dependencies in paediatric cancers by applying CRISPR-Cas technology and to
identify novel therapeutic treatments in overcoming these malignancies complemented by drug
screening data datasets to identify therapeutically actionable targets. With this experiment, the
pooled CRISPR screening pipeline is implemented which involves cultivation of rare-low survival
paediatric cancers (i.e., Neuroblastoma, High-Grade Gliomas, Sarcomas, and Atypical
Teratoid/Rhabdoid Tumour), Cas9 transduction using lentiviral vectors, green fluorescent protein
(GFP) validation, multiplicity of infection (MOI) assay, and culturing pooled CRISPR-Cas9
loss-of-function screened cell lines. Result has shown that BCL2L1 and RAC1 were found to be hit
tumour-specific genes in neuroblastoma and sarcoma, respectively. However, no statistical
differences were found when compared to other cohorts. Furthermore, MYCN overexpression and no
MYCN overexpression neuroblastoma cohorts were compared and 5 MYCN overexpression specific hit
genes were found (CDK4, EWSR1, DHFR, SDHA, and SMARCA4). Three of those genes are targetable
according to drug screening datasets and some drugs were found to be effective in addressing both
MYCN overexpression and no MYCN overexpression neuroblastoma cohorts. Further validation
should be done in the future (e.g., arrayed screening) of each gene to understand their molecular
pathways in their tumour-specific environment.

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Availability

Detail Information

Series Title

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Call Number

BT 24-005

Publisher

i3L, Jakarta : i3L, Jakarta., 2024

Collation

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Language

English

ISBN/ISSN

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Classification

NONE

Content Type

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Media Type

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Carrier Type

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Edition

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Subject(s)

Targeted therapy
Precision Medicine
Paediatric Cancer
Brain Cancer
CRISPR screens

Specific Detail Info

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Statement of Responsibility

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