Dissecting the impact of chimeric antigen receptor expression levels on T cell behaviour and response
Chimeric antigen receptor (CAR) T cells have heralded in a new age in cancer treatment. CAR T therapy has demonstrated remarkable responses in cancer patients of blood cancers. The therapy's success lies in the expression of a chimeric antigen receptor (CAR) that reprograms and redirects T cell...
| Hoofdauteur: | |
|---|---|
| Andere auteurs: | |
| Formaat: | Thesis |
| Taal: | English |
| Gepubliceerd in: |
2022
|
| Onderwerpen: |
| _version_ | 1826309463218323456 |
|---|---|
| author | Ramos, AS |
| author2 | Fulga, T |
| author_facet | Fulga, T Ramos, AS |
| author_sort | Ramos, AS |
| collection | OXFORD |
| description | Chimeric antigen receptor (CAR) T cells have heralded in a new age in cancer treatment. CAR T therapy has demonstrated remarkable responses in cancer patients of blood cancers. The therapy's success lies in the expression of a chimeric antigen receptor (CAR) that reprograms and redirects T cells to specifically target and attack cancer cells. However, CAR T therapy has faced numerous challenges with attempts to translate this success into solid cancers. Issues related to on-target, off-tumour toxicity, tonic signalling, and T cell dysfunction have limited numerous efforts. However, recent evidence demonstrated that CAR expression levels are a critical determinant in T cell activity and therapeutic efficacy – even more so in mediating T cell exhaustion. While insightful, this work assesses only two levels of CAR expression. Thus, there is a notable, outstanding gap in thoroughly understanding how CAR expression levels impact T cell function and response. In this thesis, I report the effects of extensive fine-tuning of CAR expression levels on the T cell transcriptome. First, I adapt microRNA silencing-mediated fine-tuners (miSFITs) technology to tune CAR expression levels in a T cell model. I leverage this model to show that tuning CAR expression correspondingly tunes CAR T cell activation. Next, I translate this technology into primary, human T cells, showing that miSFITs robustly and precisely tune CAR expression. I then demonstrate how tuned CAR expressions controls the expression of CD69 to stepwise levels. Finally, I use single-cell RNA-sequencing to comprehensively dissect the effects of tuning CAR expression in T cell responses, generating a transcriptomic atlas against step-wise levels of CAR expression. I use this atlas to first demonstrate how tuning CAR expression impacts several genes related to T cell activation, dysfunction, and memory. Using gene signatures, I demonstrate how CAR expression can be optimised to mitigate T cell dysfunction yet spare T cell activation and memory. Embarking on an unbiased discovery analysis, I identify genes, regulons, and signaling pathways that are influenced by CAR expression, providing a novel view on T cell dysfunction. My findings underscore the important role CAR expression levels have on T cell response and provide a promising avenue to address pressing, longstanding challenges hindering the success of CAR-T therapies. |
| first_indexed | 2024-03-07T07:36:09Z |
| format | Thesis |
| id | oxford-uuid:3e205d70-e3ca-45b7-982f-22d35ec7aa4d |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:36:09Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | oxford-uuid:3e205d70-e3ca-45b7-982f-22d35ec7aa4d2023-03-09T09:46:51ZDissecting the impact of chimeric antigen receptor expression levels on T cell behaviour and responseThesishttp://purl.org/coar/resource_type/c_db06uuid:3e205d70-e3ca-45b7-982f-22d35ec7aa4dT cellsSystems biologyImmunologyGenomicsImmunotherapyComputational biologySynthetic biologyEnglishHyrax Deposit2022Ramos, ASFulga, TMichaels, YSauka-Spengler, TChimeric antigen receptor (CAR) T cells have heralded in a new age in cancer treatment. CAR T therapy has demonstrated remarkable responses in cancer patients of blood cancers. The therapy's success lies in the expression of a chimeric antigen receptor (CAR) that reprograms and redirects T cells to specifically target and attack cancer cells. However, CAR T therapy has faced numerous challenges with attempts to translate this success into solid cancers. Issues related to on-target, off-tumour toxicity, tonic signalling, and T cell dysfunction have limited numerous efforts. However, recent evidence demonstrated that CAR expression levels are a critical determinant in T cell activity and therapeutic efficacy – even more so in mediating T cell exhaustion. While insightful, this work assesses only two levels of CAR expression. Thus, there is a notable, outstanding gap in thoroughly understanding how CAR expression levels impact T cell function and response. In this thesis, I report the effects of extensive fine-tuning of CAR expression levels on the T cell transcriptome. First, I adapt microRNA silencing-mediated fine-tuners (miSFITs) technology to tune CAR expression levels in a T cell model. I leverage this model to show that tuning CAR expression correspondingly tunes CAR T cell activation. Next, I translate this technology into primary, human T cells, showing that miSFITs robustly and precisely tune CAR expression. I then demonstrate how tuned CAR expressions controls the expression of CD69 to stepwise levels. Finally, I use single-cell RNA-sequencing to comprehensively dissect the effects of tuning CAR expression in T cell responses, generating a transcriptomic atlas against step-wise levels of CAR expression. I use this atlas to first demonstrate how tuning CAR expression impacts several genes related to T cell activation, dysfunction, and memory. Using gene signatures, I demonstrate how CAR expression can be optimised to mitigate T cell dysfunction yet spare T cell activation and memory. Embarking on an unbiased discovery analysis, I identify genes, regulons, and signaling pathways that are influenced by CAR expression, providing a novel view on T cell dysfunction. My findings underscore the important role CAR expression levels have on T cell response and provide a promising avenue to address pressing, longstanding challenges hindering the success of CAR-T therapies. |
| spellingShingle | T cells Systems biology Immunology Genomics Immunotherapy Computational biology Synthetic biology Ramos, AS Dissecting the impact of chimeric antigen receptor expression levels on T cell behaviour and response |
| title | Dissecting the impact of chimeric antigen receptor expression levels on T cell behaviour and response |
| title_full | Dissecting the impact of chimeric antigen receptor expression levels on T cell behaviour and response |
| title_fullStr | Dissecting the impact of chimeric antigen receptor expression levels on T cell behaviour and response |
| title_full_unstemmed | Dissecting the impact of chimeric antigen receptor expression levels on T cell behaviour and response |
| title_short | Dissecting the impact of chimeric antigen receptor expression levels on T cell behaviour and response |
| title_sort | dissecting the impact of chimeric antigen receptor expression levels on t cell behaviour and response |
| topic | T cells Systems biology Immunology Genomics Immunotherapy Computational biology Synthetic biology |
| work_keys_str_mv | AT ramosas dissectingtheimpactofchimericantigenreceptorexpressionlevelsontcellbehaviourandresponse |