Preclinical platforms to study therapeutic efficacy of human γδ T cells
Lingling Ou
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Stomatological Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorHuaishan Wang
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorHui Huang
The First Affiliated Hospital of Jinan University, Guangzhou, China
Search for more papers by this authorZhiyan Zhou
The First Affiliated Hospital of Jinan University, Guangzhou, China
Search for more papers by this authorQiang Lin
The First Affiliated Hospital of Jinan University, Guangzhou, China
Search for more papers by this authorYeye Guo
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorTara Mitchell
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorAlexander C. Huang
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorGiorgos Karakousis
Department of Surgery, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorLynn Schuchter
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorRavi Amaravadi
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorWei Guo
Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorJoseph Salvino
Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania, USA
Search for more papers by this authorMeenhard Herlyn
Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania, USA
Search for more papers by this authorCorresponding Author
Xiaowei Xu
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Correspondence
Dr. Xiaowei Xu, Department of Pathology and Laboratory Medicine, University of Pennsylvania, 6th Founders Building, 3400 Spruce Street, Philadelphia, PA 19104, USA.
Email: [email protected]
Search for more papers by this authorLingling Ou
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Stomatological Hospital, Southern Medical University, Guangzhou, China
Search for more papers by this authorHuaishan Wang
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorHui Huang
The First Affiliated Hospital of Jinan University, Guangzhou, China
Search for more papers by this authorZhiyan Zhou
The First Affiliated Hospital of Jinan University, Guangzhou, China
Search for more papers by this authorQiang Lin
The First Affiliated Hospital of Jinan University, Guangzhou, China
Search for more papers by this authorYeye Guo
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorTara Mitchell
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorAlexander C. Huang
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorGiorgos Karakousis
Department of Surgery, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorLynn Schuchter
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorRavi Amaravadi
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorWei Guo
Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Search for more papers by this authorJoseph Salvino
Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania, USA
Search for more papers by this authorMeenhard Herlyn
Molecular and Cellular Oncogenesis Program, The Wistar Institute, Philadelphia, Pennsylvania, USA
Search for more papers by this authorCorresponding Author
Xiaowei Xu
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA
Correspondence
Dr. Xiaowei Xu, Department of Pathology and Laboratory Medicine, University of Pennsylvania, 6th Founders Building, 3400 Spruce Street, Philadelphia, PA 19104, USA.
Email: [email protected]
Search for more papers by this authorGW and XX are scientific co-founders of CureBiotech and Exio Biosciences.
Abstract
Background
Gamma delta (γδ) T lymphocytes are promising candidate for adoptive T cell therapy, however, their treatment efficacy is not satisfactory. Vδ2 T cells are unique to primates and few suitable models are available to assay their anti-tumour function.
Methods
We tested human γδ T cell activation, tumour infiltration, and tumour-killing in four three-dimensional (3D) models, including unicellular, bicellular and multicellular melanoma spheroids, and patient-derived melanoma organoids. We studied the effects of checkpoint inhibitors on γδ T cells and performed a small molecule screen using these platforms.
Results
γδ T cells rapidly responded to melanoma cells and infiltrated melanoma spheroids better than αβ T cells in PBMCs. Cancer-associated fibroblasts (CAFs) in bicellular spheroids, stroma cells in multicellular melanoma spheroids and inhibitory immune cells in organoids significantly inhibited immune cell infiltrates including γδ T cells and lessened their cytotoxicity to tumour cells. Tumour-infiltrating γδ T cells showed exhausted immunophenotypes with high checkpoints expression (CTLA-4, PD-1 and PD-L1). Immune checkpoint inhibitors increased γδ T cell infiltration of 3D models and killing of melanoma cells in all four 3D models. Our small molecule screen assay and subsequent mechanistic studies demonstrated that epigenetic modifiers enhanced the chemotaxis and cytotoxicity of γδ T cells through upregulating MICA/B, inhibiting HDAC6/7 pathway and downregulating the levels of PD-L1 and PD-L2 in CAFs and tumour cells. These compounds increased CXCR4 and CD107a expression, IFN-γ production and decreased PD-1 expression of γδ T cells.
Conclusions
Tumour-infiltrating γδ T cells show exhausted immunophenotypes and limited anti-tumour capacity in melanoma 3D models. Checkpoint inhibitors and epigenetic modifiers enhance anti-tumour functions of γδ T cells. These four 3D models provided valuable preclinical platforms to test γδ T cell functions for immunotherapy.
Supporting Information
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| ctm2814-sup-0007-figureS7.jpg1.1 MB | Supporting Information |
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