T cells engineered to express a T-cell receptor specific for glypican-3 recognize and kill hepatoma cells in vitro and in mice

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Item Type:	Article
Title:	T cells engineered to express a T-cell receptor specific for glypican-3 recognize and kill hepatoma cells in vitro and in mice
Creators Name:	Dargel, C., Bassani-Sternberg, M., Hasreiter, J., Zani, F., Bockmann, J.H., Thiele, F., Bohne, F., Wisskirchen, K., Wilde, S., Sprinzl, M.F., Schendel, D.J., Krackhardt, A.M., Uckert, W., Wohlleber, D., Schiemann, M., Stemmer, K., Heikenwälder, M., Busch, D.H., Richter, G., Mann, M. and Protzer, U.
Abstract:	BACKGROUND & AIMS: Cancer therapies are being developed based on our ability to direct T cells against tumor antigens. Glypican 3 (GPC3) is expressed by 75% of all hepatocellular carcinomas (HCC) but not in healthy liver tissue or other organs. We aimed to generate T cells with GPC3-specific receptors that recognize HCC and used them to eliminate GPC3-expressing xenograft tumors grown from human HCC cells in mice. METHODS: We used mass spectrometry and to obtain a comprehensive peptidome from GPC3-expressing hepatoma cells after immune-affinity purification of HLA-A2, and used bioinformatics to identify immunodominant peptides. To circumvent GPC3-tolerance resulting from fetal expression, dendritic cells from HLA-A2 negative donors were co-transfected with GPC3 and HLA-A2 RNA to stimulate and expand antigen-specific T cells. RESULTS: Peptide GPC3367 was identified as a predominant peptide on HLA-A2. We used A2-GPC3367 multimers to detect, select for, and clone GPC3-specific T cells. These clones bound the A2-GPC3367 multimer and secreted interferon-gamma when cultured with GPC3367, but not with control peptide loaded cells. By genomic sequencing of these T-cell clones, we identified a gene encoding a dominant T-cell receptor. The gene was cloned, the sequence was codon optimized and expressed from a retroviral vector. Primary CD8+ T cells that expressed the transgenic T-cell receptor specifically bound GPC3367 on HLA-A2. These T cells killed GPC3-expressing hepatoma cells in culture and slowed growth of HCC xenograft tumors in mice. CONCLUSION: We identified a GPC3367-specific T-cell receptor. Expression of this receptor by T cells allows them to recognize and kill GPC3-positive hepatoma cells. This finding could be used to advance development of adoptive T-cell therapy for HCC.
Keywords:	Cancer Immunotherapy, Tumor-Associated Antigens, Liver Cancer, Immune Response, Animals, Mice
Source:	Gastroenterology
ISSN:	0016-5085
Publisher:	Elsevier / Saunders
Volume:	149
Number:	4
Page Range:	1042-1052
Date:	October 2015
Official Publication:	https://doi.org/10.1053/j.gastro.2015.05.055
PubMed:	View item in PubMed

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