Functional genomic screening reveals regulators of galectin-1 intracellular trafficking with applications to cellular delivery of new therapeutic modalities
Abstract/Contents
- Abstract
- Many novel modalities like small-interfering RNA (siRNA), CRISPR, and recombinant proteins have revolutionized biological research, but their applicability in the clinic remains limited. These massive and often highly charged molecules cannot cross the lipophilic cell membrane, and methods for delivering these molecules to the cell interior in a clinical setting are a key bottleneck to their translation. As introduced in Chapter 1, nearly all methods for delivering biomolecules to cells rely in some way on a cell's own trafficking machinery and the glycan-binding protein galectin-1 is able to traffic from the cell surface to the cell interior via an unknown mechanism. Thus, to address this challenge, we shed new light on the trafficking of galectin-1 and explore the possibility of hijacking this trafficking system to deliver therapeutic molecules. Functional genomics represents a powerful new set of tools for mechanistic investigation of biological phenomena in an unbiased manner. In Chapter 2, we explore and prototype selection methodologies that would enable us to leverage genome-wide CRISPR screening to study galectin-1 trafficking. Chapter 3 describes the use of a galectin-1-toxin conjugate, modelled on antibody-drug conjugates, as a selection tool in a genome-wide CRISPR screen and discusses the resulting data. Next, in Chapter 4, we validate the results of the screen and perform a deeper investigation into the pathways followed by exogenous galectin-1 when it enters cells. We discover that galectin-1 interacts with the endosome-lysosome trafficking receptor sortilin in a glycan-dependent manner and that this interaction regulates galectin-1 trafficking to the lysosome. Finally, in Chapter 5, we show that the galectin-1 trafficking pathway can be exploited for delivery of a functional siRNA. Taken together, our work illuminates the mechanisms by which galectin-1 is internalized by cells and suggests a new strategy for intracellular drug delivery via galectin-1 conjugation.
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 | 2024; ©2024 |
| Publication date | 2023; 2023 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Donnelly, Justin |
|---|---|
| Degree supervisor | Bertozzi, Carolyn R, 1966- |
| Thesis advisor | Bertozzi, Carolyn R, 1966- |
| Thesis advisor | Bassik, Michael |
| Thesis advisor | Long, Jonathan Z |
| Degree committee member | Bassik, Michael |
| Degree committee member | Long, Jonathan Z |
| Associated with | Stanford University, School of Humanities and Sciences |
| Associated with | Stanford University, Department of Chemistry |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Justin Donnelly. |
|---|---|
| Note | Submitted to the Department of Chemistry. |
| Thesis | Thesis Ph.D. Stanford University 2024. |
| Location | https://purl.stanford.edu/cs733vr9198 |
Access conditions
- Copyright
- © 2024 by Justin Donnelly
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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