Single-Cell Analysis of CAR-T Cell Activation Reveals A Mixed T H 1/T H 2 Response Independent of Differentiation
Year: 2019
Type: article
Abstract: The activation mechanism of chimeric antigen receptor (CAR)-engineered T cells may differ substantially from T cells carrying native T cell receptor, but this difference remains poorly understood. We present the first comprehensive portrait of single-cell level transcriptional and cytokine signatures of anti-CD19/4-1BB/CD28/CD3ζ CAR-T cells upon antigen-specific stimulation. Both CD4+ helper T (TH) cells and CD8+ cytotoxic CAR-T cells are equally effective in directly killing target tumor cells and their cytotoxic activity is associated with the elevation of a range of TH1 and TH2 signature cytokines, e.g., interferon γ, tumor necrotic factor α, interleukin 5 (IL5), and IL13, as confirmed by the expression of master transcription factor genes TBX21 and GATA3. However, rather than conforming to stringent TH1 or TH2 subtypes, single-cell analysis reveals that the predominant response is a highly mixed TH1/TH2 function in the same cell. The regulatory T cell activity, although observed in a small fraction of activated cells, emerges from this hybrid TH1/TH2 population. Granulocyte-macrophage colony stimulating factor (GM-CSF) is produced from the majority of cells regardless of the polarization states, further contrasting CAR-T to classic T cells. Surprisingly, the cytokine response is minimally associated with differentiation status, although all major differentiation subsets such as naïve, central memory, effector memory, and effector are detected. All these suggest that the activation of CAR-engineered T cells is a canonical process that leads to a highly mixed response combining both type 1 and type 2 cytokines together with GM-CSF, supporting the notion that polyfunctional CAR-T cells correlate with objective response of patients in clinical trials. This work provides new insights into the mechanism of CAR activation and implies the necessity for cellular function assays to characterize the quality of CAR-T infusion products and monitor therapeutic responses in patients.
Institutions: Yale Cancer Center, Yale University
Language: English
Topic: CAR-T cell therapy research
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
Estimated APC paid: $2,200
Funders: National Science Foundation, Yale University, Yale Cancer Center, National Institutes of Health, National Science Foundation of Sri Lanka, National Cancer Institute, National Institute of Diabetes and Digestive and Kidney Diseases, Division of Chemical, Bioengineering, Environmental, and Transport Systems
Awards: Evolution and Treatment Response of Brain, Breast, and Hematologic Malignancies, CAREER: Revisiting the Central Dogma at the Genome Scale and in Single Cells, G3634416345, Systems analysis of phenotypic switch in control of cancer invasion, G6671297155, Yale Cooperative Center of Excellence in Hematology, G9000142786, G9000320151
OpenAlex ID: W2952737707