1. Designed improvement to T-cell immunotherapy by multidimensional single cell profiling
- Author
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Rasindu B. Rajanayake, Ivan Liadi, Patrick Hwu, Irfan N Bandey, Jay R Adolacion, Arash Saeedi, Harjeet Singh, Navin Varadarajan, Zheng You, Gabrielle Romain, Melisa Martinez Paniagua, Xingyue An, and Laurence J.N. Cooper
- Subjects
0301 basic medicine ,Cytotoxicity, Immunologic ,Cancer Research ,medicine.medical_treatment ,T-Lymphocytes ,Cell ,immunologic techniques ,Mice, SCID ,Immunotherapy, Adoptive ,Cell therapy ,0302 clinical medicine ,Mice, Inbred NOD ,Immunology and Allergy ,Hepatitis A Virus Cellular Receptor 2 ,RC254-282 ,Ovarian Neoplasms ,Leukemia ,Receptors, Chimeric Antigen ,CD137 ,Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Natural killer T cell ,medicine.anatomical_structure ,Phenotype ,Oncology ,030220 oncology & carcinogenesis ,Molecular Medicine ,CD8-positive T-lymphocytes ,Female ,immunotherapy ,Single-Cell Analysis ,Immunology ,Biology ,cell engineering ,adoptive ,CD19 ,Immunophenotyping ,03 medical and health sciences ,Antigen ,medicine ,Animals ,Humans ,Pharmacology ,Immune Cell Therapies and Immune Cell Engineering ,Gene Expression Profiling ,Immunotherapy ,Xenograft Model Antitumor Assays ,Chimeric antigen receptor ,030104 developmental biology ,4-1BB Ligand ,Cancer research ,biology.protein ,K562 Cells ,Transcriptome - Abstract
BackgroundAdoptive cell therapy based on the infusion of chimeric antigen receptor (CAR) T cells has shown remarkable efficacy for the treatment of hematologic malignancies. The primary mechanism of action of these infused T cells is the direct killing of tumor cells expressing the cognate antigen. However, understanding why only some T cells are capable of killing, and identifying mechanisms that can improve killing has remained elusive.MethodsTo identify molecular and cellular mechanisms that can improve T-cell killing, we utilized integrated high-throughput single-cell functional profiling by microscopy, followed by robotic retrieval and transcriptional profiling.ResultsWith the aid of mathematical modeling we demonstrate that non-killer CAR T cells comprise a heterogeneous population that arise from failure in each of the discrete steps leading to the killing. Differential transcriptional single-cell profiling of killers and non-killers identified CD137 as an inducible costimulatory molecule upregulated on killer T cells. Our single-cell profiling results directly demonstrate that inducible CD137 is feature of killer (and serial killer) T cells and this marks a different subset compared with the CD107apos (degranulating) subset of CAR T cells. Ligation of the induced CD137 with CD137 ligand (CD137L) leads to younger CD19 CAR T cells with sustained killing and lower exhaustion. We genetically modified CAR T cells to co-express CD137L, in trans, and this lead to a profound improvement in anti-tumor efficacy in leukemia and refractory ovarian cancer models in mice.ConclusionsBroadly, our results illustrate that while non-killer T cells are reflective of population heterogeneity, integrated single-cell profiling can enable identification of mechanisms that can enhance the function/proliferation of killer T cells leading to direct anti-tumor benefit.
- Published
- 2021