Applications of CRISPR-Cas9 Technology to Genome Editing in Glioblastoma Multiforme

Abstract

Glioblastoma multiforme (GBM) is an aggressive malignancy of the brain and spinal cord with a poor life expectancy. The low survivability of GBM patients can be attributed, in part, to its heterogeneity and the presence of multiple genetic alterations causing rapid tumor growth and resistance to conventional therapy. The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associated (Cas) nuclease 9 (CRISPR-Cas9) system is a cost-effective and reliable gene editing technology, which is widely used in cancer research. It leads to novel discoveries of various oncogenes that regulate autophagy, angiogenesis, and invasion and play important role in pathogenesis of various malignancies, including GBM. In this review article, we first describe the principle and methods of delivery of CRISPR-Cas9 genome editing. Second, we summarize the current knowledge and major applications of CRISPR-Cas9 to identifying and modifying the genetic regulators of the hallmark of GBM. Lastly, we elucidate the major limitations of current CRISPR-Cas9 technology in the GBM field and the future perspectives. CRISPR-Cas9 genome editing aids in identifying novel coding and non-coding transcriptional regulators of the hallmarks of GBM particularly in vitro, while work using in vivo systems requires further investigation.

Systems Medicine for Precise Targeting of Glioblastoma

“…There is an interesting review that discussed the application of CRISPR-Cas9 tool to identifying genes associated with self-regeneration, growth, angiogenesis, inflammation, apoptosis, cell migration, and invasion factors. Moreover, CRISPR-Cas9 screens are utilized to identify new biomarkers, oncogenic drivers, and cause of chemotherapy resistance in vivo or in vitro [ 125 ]. In addition, CRISPR-Cas9 genome editing can be applied to detect novel coding and non-coding transcriptional regulators of the GBM in vitro, although the application of CRISPR-Cas systems in vivo systems requires further research and development.…”

Advanced Bioinformatics Analysis and Genetic Technologies for Targeting Autophagy in Glioblastoma Multiforme

“…Now going beyond its ten-year mark since the publication of the landmark paper in Science first describing how CRISPR-Cas9 technology can be used for precise gene editing, much investigation has taken place with respect to the identification of the function of various genes related to the regulation of autophagy and the elucidation of applications to GBM characterization and therapy [ 118 ]. However, the use of CRISPR-Cas9 technology for the modification of ARGs in GBM is still incredibly limited [ 13 ]. Leveraging CRISPR-Cas9 modified organoid models and CAR-T cells with decreased sensitivity to immunosuppressive factors, a TME and GBM model conducive to functional analysis of ARGs may reveal novel targets that will potentiate the therapeutic action of TMZ intended specifically for induction of apoptosis.…”

“…Limitations of the efficacy of current therapeutics in GBM and the presence of processes that confer therapeutic resistance, such as cytoprotective autophagy, have called for the development of novel therapeutic options to combat the progression of this deadly disease and prevent tumor recurrence. Current avenues of investigation include the identification of novel biomarkers to detect levels of autophagy, genome editing with advanced technology such as the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas) or CRISPR-Cas9 system to mitigate chemotherapeutic resistance, the use of targeted microRNA (miRNA) to turn off genes that promote autophagy, and employment of the plant-derived bioflavonoids to inhibit autophagy and potentiate the therapeutic action of TMZ in GBM [ 13 , 14 ]. The focus of this review article is to provide a broad overview of the interaction between autophagy and GBM progression and the relationship between TMZ and autophagy, as well as highlight the potential perceived and limitations of current therapeutics being developed ( Figure 1 ).…”

Friends with Benefits: Chemokines, Glioblastoma-Associated Microglia/Macrophages, and Tumor Microenvironment

“…The most frequently-used gene-editing technology, CRISPR/Cas9, can be used for single target knockout or as a screening tool, using sgRNA libraries, to collect data on genetic regulators of invasion [ 277 ], radiation resistance [ 278 ], temozolomide resistance [ 279 ], interaction to the extracellular matrix [ 280 ], apoptosis, or autophagy in GBM (reviewed in [ 281 ]). Genome-wide gene editing screens were proven effective for highlighting gene hits outside of core genes and regular pathways described for GBM, such as Wee-1 kinase [ 282 ], which prevents mitotic entry, currently targeted by orally available inhibitors and tested in clinical trials [ 283 ].…”