Identification of synergistic cancer drug combinations and host modifiers of legionella pneumophila pathogenesis using CRISPR-Cas9 screens
Abstract/Contents
- Abstract
- The recent development of CRISPR-Cas9 technology has enabled robust high-throughput gene deletion screening as a powerful tool for discovery in mammalian cells. In this work, I present two diverse applications of CRISPR-Cas9 pooled high-throughput functional screening. First, I describe the development of a CRISPR-based double knockout system, which we use to screen for synergistic cancer drug combinations. While combination therapies can be an effective way to reduce drug resistance to targeted single-drug therapies, conventional methods of testing drug combinations have poor scalability. Using the CRISPR double knockout system, we generate a large-scale mammalian genetic interaction map comprised of ~490,000 double-sgRNA combinations corresponding to 21,321 drug target pairs in a search for synergistically lethal drug combinations in leukemia cells. We discover combination drug therapy targeting BCL2L1 (BCL-XL) and MCL1 as a promising therapy for leukemia with highly synergistic cancer-specific effects. Next, I describe a second application of CRISPR screening towards exploring the biology of a unique intracellular bacterial pathogen Legionella pneumophila that is able to drastically disrupt host cell processes during infection. I perform a genome-wide single knockout screen and secondary targeted screens to discover hundreds of host factors that mediate the pathogenesis of L. pneumophila during infection of human monocyte and macrophage cells. In addition to identifying two previously uncharacterized genes, C1ORF43 and KIAA1109 as novel regulators of phagocytosis, we highlight the GTPase RAB10 and its chaperone RABIF as key regulators of L. pneumophila replication and vacuole maturation. Collectively, this work demonstrates the power of high-throughput genetic screens for discovering novel drug combinations for cancer and improving our understanding of L. pneumophila host-pathogen interactions.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2019; ©2019 |
| Publication date | 2019; 2019 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Jeng, Edwin Ethan |
|---|---|
| Degree supervisor | Bassik, Michael |
| Thesis advisor | Bassik, Michael |
| Thesis advisor | Attardi, Laura |
| Thesis advisor | Jackson, Peter K. (Peter Kent) |
| Thesis advisor | Sage, Julien |
| Degree committee member | Attardi, Laura |
| Degree committee member | Jackson, Peter K. (Peter Kent) |
| Degree committee member | Sage, Julien |
| Associated with | Stanford University, Cancer Biology Program. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Edwin Jeng. |
|---|---|
| Note | Submitted to the Cancer Biology Program. |
| Thesis | Thesis Ph.D. Stanford University 2019. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2019 by Edwin Ethan Jeng
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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