Engineering novel leukemia inhibitory factor inhibitors as cancer therapeutics
Abstract/Contents
- Abstract
- Pancreatic ductal adenocarcinoma (PDAC) is a devastating form of cancer, with the lowest five-year survival rate among all cancers of only 9%. Poor survival is due to late detection, limited ability to surgically resect disease, and a dearth of targeted, effective therapies. As covered in Chapter 1, numerous recent studies have identified the leukemia inhibitory factor (LIF) as a promising therapeutic target in PDAC. LIF, a member of the IL-6 family of cytokines, drives proliferation, survival, and stem-ness cellular programming via signaling though the heterodimeric receptor pair of LIF receptor (LIFR) and glycoprotein 130 (gp130). While numerous potential inhibitors of LIF have been developed, because of the high affinity of LIF for the LIFR/gp130 complex, and the shared nature of cytokine receptors, non-toxic blockade of LIF activity remains challenging. The development of novel and effective LIF inhibitors is thus sorely needed as promising therapeutics to use in the fight against PDAC. In this dissertation, I will describe our work to engineer a novel and potent type of LIF inhibitor known as a ligand trap. Ligand traps have recently emerged as a new class of inhibitors, based on the principle of using a soluble form of a receptor to bind and sequester soluble ligands from activating cellular signaling. To inhibit LIF, we engineered both LIFR and gp130 for heightened affinity to LIF using the combinatorial protein engineering techniques of yeast-surface display and directed evolution. In Chapter 2, I cover the basic principles behind determining affinity and performing affinity maturation using yeast-surface display, including numerous yeast-based characterization assays, critical for determining therapeutic efficacy. Chapter 3 describes the engineering and production of a soluble version of LIFR, called eLIFR-Fc. As a ligand trap, eLIFR-Fc possessed many desirable therapeutic properties, including very high affinity for LIF, direct competition with LIF's receptors, blockade of LIF derived signaling in cancer cells, and abrogation of PDAC growth in vivo. Chapter 4 covers potential improvements to the eLIFR-Fc design, via fusion of engineered gp130, to create a potent engineered fusion protein (eFusion-Fc). By inclusion of both of LIF's receptors, the eFusion-Fc protein eliminated LIF signaling via LIFR and gp130 in cancer cells. These engineered ligand traps are extremely potent LIF inhibitors and add to the legacy of ligand traps as an emerging class of therapeutics, while offering hope as an effective PDAC treatment.
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 | 2020; ©2020 |
| Publication date | 2020; 2020 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Hunter, Sean Alexander |
|---|---|
| Degree supervisor | Cochran, Jennifer R |
| Thesis advisor | Cochran, Jennifer R |
| Thesis advisor | Jackson, Peter K. (Peter Kent) |
| Thesis advisor | Kim, Peter, 1958- |
| Thesis advisor | Sage, Julien |
| Degree committee member | Jackson, Peter K. (Peter Kent) |
| Degree committee member | Kim, Peter, 1958- |
| Degree committee member | Sage, Julien |
| Associated with | Stanford University, Cancer Biology Program |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Sean Alexander Hunter. |
|---|---|
| Note | Submitted to the Cancer Biology Program. |
| Thesis | Thesis Ph.D. Stanford University 2020. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Sean Alexander Hunter
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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