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| Status |
Public on Jan 21, 2021 |
| Title |
mRNA profiling on CD19-directed chimeric antigen receptor T (CART19) cells with and without platelet-poor plasma from untreated chronic lymphocytic leukemia (CLL) patients (CLL-derived extracellular vesicles) |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Chimeric antigen receptor (CAR) T cell therapy has yielded unprecedented outcomes in some patients with hematological malignancies; however, inhibition by the tumor microenvironment has prevented the broader success of CART cell therapy. We used chronic lymphocytic leukemia (CLL) as a model to investigate the interactions between the tumor microenvironment and CART cells. CLL is characterized by an immunosuppressive microenvironment, an abundance of systemic extracellular vesicles (EVs), and a relatively lower durable response rate to CART cell therapy. In this study, we characterized plasma EVs from untreated CLL patients and identified their leukemic cell origin. CLL-derived EVs were able to induce a state of CART cell dysfunction characterized by phenotypical, functional, and transcriptional changes of exhaustion. We demonstrate that, specifically, PD-L1+ CLL-derived EVs induce CART cell exhaustion. In conclusion, we identify an important mechanism of CART cell exhaustion induced by EVs from CLL patients.
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| Overall design |
RNA sequencing was performed on antigen-stimulated CART19 co-cultured with three biological replicates of CLL-derived EVs or alone.
CART19 and irradiated JeKo-1 were co-cultured at a 1:1 ratio for 24 hours with CLL-derived EVs at 10:1 and 1:1 EV:CART19 ratios. Three biological replicates of CLL-derived EVs were included as well as antigen-stimulated and unstimulated CART19 controls without EVs. CART19 were isolated using magnetic sorting with CD4 and CD8 microbeads. RNA was isolated from the CART19 cells using QIAGEN miRNeasy Micro Kit.
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| Contributor(s) |
Cox M, Lucien F, Kay N, Kenderian S |
| Citation(s) |
33388419 |
| |
| Submission date |
Mar 16, 2020 |
| Last update date |
Jan 24, 2021 |
| Contact name |
Saad Kenderian |
| E-mail(s) |
[email protected]
|
| Organization name |
Mayo Clinic
|
| Department |
Hematology Research
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| Lab |
T Cell Engineering
|
| Street address |
200 First Street SW
|
| City |
Rochester |
| State/province |
MN |
| ZIP/Postal code |
55905 |
| Country |
USA |
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| Platforms (1) |
| GPL20301 |
Illumina HiSeq 4000 (Homo sapiens) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA612847 |
| SRA |
SRP252970 |
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