Identification of genetic regulators of antibody-drug conjugate toxicity using CRISPR-Cas9 screens
Abstract/Contents
- Abstract
- Antibody-drug conjugates (ADCs) selectively deliver highly toxic chemotherapeutic agents to target antigen-expressing cells and have become an important cancer treatment in recent years. However, the molecular mechanisms by which ADCs are internalized and activated within cells remain unclear. Here we use CRISPR-Cas9 screens to identify genes that control the toxicity of ADCs targeting different cancer antigens and cell types. Our results demonstrate critical roles for a range of known and novel intracellular trafficking regulators in ADC toxicity. We identify and characterize C18orf8/RMC1 as a regulator of ADC toxicity through its role in endosomal maturation. Through comparative analysis of CRISPR screens with ADCs bearing a noncleavable linker versus a cleavable valine-citrulline (VC) linker, we show that a subset of late endo/lysosomal and retrograde trafficking regulators selectively influence toxicity of noncleavable linker ADCs. We further show that cleavable VC linkers are rapidly processed upon internalization and therefore surprisingly appear to bypass the requirement of lysosomal delivery. Lastly, we show that depletion of sialic acids sensitizes cells to anti-CD22 and anti-Her2 ADCs by increasing the rate of ADC lysosomal delivery. In contrast, deletion of sialic acid biosynthesis genes protects cells from anti-CD30 ADCs, suggesting that proper sialylation is required for CD30 cell surface-expression. Together, these results reveal novel regulators of endolysosomal trafficking, provide important insights to guide future ADC design, and identify candidate combination therapy targets as well as potential mechanisms of ADC resistance.
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 | Tsui, Chung Yin Kimberly |
|---|---|
| Degree supervisor | Bassik, Michael |
| Thesis advisor | Bassik, Michael |
| Thesis advisor | Brunet, Anne, 1972- |
| Thesis advisor | Gitler, Aaron D |
| Thesis advisor | Pfeffer, Suzanne |
| Thesis advisor | Wyss-Coray, Anton |
| Degree committee member | Brunet, Anne, 1972- |
| Degree committee member | Gitler, Aaron D |
| Degree committee member | Pfeffer, Suzanne |
| Degree committee member | Wyss-Coray, Anton |
| Associated with | Stanford University, Department of Genetics. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | C. Kimberly Tsui. |
|---|---|
| Note | Submitted to the Department of Genetics. |
| Thesis | Thesis Ph.D. Stanford University 2019. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2019 by Chung Yin Kimberly Tsui
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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