Using CRISPR Screens to Identify Genetic Vulnerabilities to Cancer Treatment
Open Access
- Author:
- Schleicher, Emily
- Graduate Program:
- Biomedical Sciences
- Degree:
- Doctor of Philosophy
- Document Type:
- Dissertation
- Date of Defense:
- February 23, 2022
- Committee Members:
- Hong-Gang Wang, Major Field Member
James Broach, Major Field Member
George Moldovan, Chair & Dissertation Advisor
Ralph Keil, Program Head/Chair
David Degraff, Outside Unit & Field Member
Katherine Aird, Special Member - Keywords:
- CRISPR
CRISPR Screen
ATM inhibitors
ATR inhibitors
PARP10
Resistance
Sensitivity
CRISPR Knockout
CRISPR Activation - Abstract:
- Identifying the correct treatment plan for cancer patients is critical to prevent disease progression and development of chemotherapeutic resistance. Although the cancer field has come a long way in treatment plan development, there is still a large subset of patients who experience resistance to chemotherapy. The DNA damage response is highly involved in cancer progression, and one of the main targets of emerging chemotherapeutics. In this dissertation, I reveal genes involved in cancer progression, along with genome-wide CRISPR-Cas9 screens that identify synthetic lethality interactions as well as potential biomarkers of resistance and sensitivity to DNA damage response inhibitors. I will present many examples of how CRISPR-Cas9 screens can be applied to advance future personalized medicine efforts in the clinic. Specifically, I will demonstrate how genome-wide CRISPR-Cas9 screening can be employed to identify induced lethality with a knockout screen that aims to identify genes that, when lost, cause cell death in PARP10 overexpression cells. Next, I reveal how both CRISPR knockout and CRISPR activation screens were applied to identify potential biomarkers of resistance to multiple ATR inhibitors. Finally, I exhibit how a CRISPR knockout screen in both wild-type and BRCA2-deficient cells successfully identified potential biomarkers of sensitivity to ATM inhibitors. My work provides comprehensive sets of targets and potential biomarkers for both oncogenes and drugs that are currently in clinical trials. Moreover, I reveal novel mechanisms of resistance and sensitivity from analysis of the top hits of the performed screens.