期刊
JOURNAL FOR IMMUNOTHERAPY OF CANCER
卷 9, 期 5, 页码 -出版社
BMJ PUBLISHING GROUP
DOI: 10.1136/jitc-2020-002328
关键词
immunotherapy; translational medical research; receptors; chimeric antigen
资金
- Stand-Up-to-Cancer Convergence 2.0 Grant
- Packard Fellowship for Science and Engineering
- Stand Up To Cancer, a program of the Entertainment Industry Foundation
- Society for Immunotherapy of Cancer
- Connecticut Regenerative Medicine Research Fund
- Li Ka Shing Foundation
- Yale Center for Genomics Analysis
Using single-cell mRNA sequencing and CITE-seq, researchers conducted multiomics characterization of CAR T cells from healthy donors and patients with acute lymphoblastic leukemia, revealing heterogeneities in transcriptional, phenotypic, functional, and metabolic profiles between donor and patient CAR T cells. Donor CAR T cells exhibited more pronounced activation levels upon CD19 stimulation.
Background Autologous T cells engineered to express a chimeric antigen receptor (CAR) specific for CD19 molecule have transformed the therapeutic landscape in patients with highly refractory leukemia and lymphoma, and the use of donor-generated allogeneic CAR T is paving the way for further breakthroughs in the treatment of cancer. However, it remains unknown how the intrinsic heterogeneities of these engineered cells mediate therapeutic efficacy and whether allogeneic products match the effectiveness of autologous therapies. Methods Using single-cell mRNA sequencing in conjunction with CITE-seq, we performed multiomics characterization of CAR T cells generated from healthy donor and patients with acute lymphoblastic leukemia. CAR T cells used in this study were manufactured at the University of Pennsylvania through lentiviral transduction with a CD19-4-1BB-CD3 zeta construct. Besides the baseline condition, we engineered NIH-3T3 cells with human CD19 or mesothelin expression to conduct ex vivo antigen-specific or non-antigen stimulation of CAR T cells through 6-hour coculture at a 1:1 ratio. Results We delineated the global cellular and molecular CAR T landscape and identified that transcriptional CAR tonic signaling was regulated by a mixture of early activation, exhaustion signatures, and cytotoxic activities. On CD19 stimulation, we illuminated the disparities of CAR T cells derived from different origins and found that donor CAR T had more pronounced activation level in correlation with the upregulation of major histocompatibility complex class II genes compared with patient CAR T cells. This finding was independently validated in additional datasets from literature. Furthermore, GM-CSF(CSF2) expression was found to be associated with functional gene productions, but it induced little impact on the CAR T activation. Conclusions Through integrated multiomics profiling and unbiased canonical pathway analyses, our results unveil heterogeneities in the transcriptional, phenotypic, functional, and metabolic profiles of donor and patient CAR T cells, providing mechanistic basis for ameliorating clinical outcomes and developing next-generation 'off- the-shelf' allogeneic products.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据