Enhancing the safety and efficacy of engineered cell therapies using protease-regulated platforms
Abstract/Contents
- Abstract
- Chimeric antigen receptors (CARs) are synthetic modular proteins that allow for directing immune cell reactivity towards a molecular target of interest, such as a cancer-associated antigen. CAR-T cells have demonstrated remarkable clinical activity in patients with refractory hematologic malignancies, leading to ten FDA-approved therapies to date. There is much hope that next-generation platforms leveraging synthetic biology and genetic engineering will expand this class of therapeutics to solid tumors, where toxicity risks and limited efficacy are major barriers to progress. However, current platforms for enhancing CAR potency are at risk for increased toxicity, while those that are designed for safety exhibit diminished potency. In this thesis, I first report on SNIP, an extensively optimized CAR ON switch that dually enhances safety and efficacy of T cells and is regulated by an FDA-approved small molecule. SNIP CARs outperformed conventional CARs in numerous orthotropic solid tumor models. Through phenotypic and functional studies, we show that enhanced activity of SNIP CARs is due to diminished T cell exhaustion and greater generation of memory T cells. In an on-target, off-tumor toxicity model we developed, we show that SNIP CAR activation can be tuned to fall within a therapeutic window in which T cell cytotoxic activity can be directed at tumor cells expressing high levels of antigen while sparing healthy tissue that expresses lower levels of antigen. Next, I report on STASH Tag, a drug regulated system for regulating the subcellular localization of a protein of interest. In order to meet the multiple obstacles facing CAR-T cells in solid tumors, next-generation smart therapies need to be multi-specific, resistant to exhaustion, and maintain their fitness in a hostile tumor microenvironment. Engineering next-generation therapies with these enhanced capabilities will require the introduction of additional genetic modules that often do not fit within the payload capacity of a single vector, or use a combination of gene engineering techniques (e.g. lentivirus + CRISPR), necessitating the use of multiple vectors. For my last thesis project, I report on STASH Select, a simple platform technology for enriching cells that have been engineered with multiple vectors using a single-step selection that results in high purity/yield and is compatible with current GMP cell manufacturing workflows and closed loop systems.
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 | 2022; ©2022 |
| Publication date | 2022; 2022 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Labanieh, Louai |
|---|---|
| Degree supervisor | Cochran, Jennifer R |
| Degree supervisor | Mackall, Crystal |
| Thesis advisor | Cochran, Jennifer R |
| Thesis advisor | Mackall, Crystal |
| Thesis advisor | Majzner, Robbie |
| Thesis advisor | Qi, Lei, (Professor of Bioengineering) |
| Degree committee member | Majzner, Robbie |
| Degree committee member | Qi, Lei, (Professor of Bioengineering) |
| Associated with | Stanford University, Department of Bioengineering |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Louai Labanieh. |
|---|---|
| Note | Submitted to the Department of Bioengineering. |
| Thesis | Thesis Ph.D. Stanford University 2022. |
| Location | https://purl.stanford.edu/jf768jh9079 |
Access conditions
- Copyright
- © 2022 by Louai Labanieh
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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