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Expansion of human ?? T cells for adoptive immunotherapy using a bisphosphonate prodrug.

ABSTRACT: Cancer immunotherapy with human ?? T cells expressing V?2V?2 T cell receptor (also termed V?9V?2) has shown promise because of their ability to recognize and kill most types of tumors in a major histocombatibility complex (MHC) -unrestricted fashion that is independent of the number of tumor mutations. In clinical trials, adoptive transfer of V?2V?2 T cells has been shown to be safe and does not require preconditioning. In this report, we describe a method for preparing highly enriched human V?2V?2 T cells using the bisphosphonate prodrug, tetrakis-pivaloyloxymethyl 2-(thiazole-2-ylamino)ethylidene-1,1-bisphosphonate (PTA). PTA stimulated the expansion of V?2V?2 cells to purities up to 99%. These levels were consistently higher than those observed after expansion with zoledronic acid, the most commonly used stimulator for clinical trials. Cell numbers also averaged more than those obtained with zoledronic acid and the expanded V?2V?2 cells exhibited high cytotoxicity against tumor cells. The high purity of V?2V?2 cells expanded by PTA increased engraftment success in immunodeficient NOG mice. Even low levels of contaminating ?? T cells resulted in some mice with circulating human ?? T cells rather than V?2V?2 cells. V?2V?2 cells from engrafted NOG mice upregulated CD25 and secreted tumor necrosis factor-? and interferon-? in response to PTA-treated tumor cells. Thus, PTA expands V?2V?2 T cells to higher purity than zoledronic acid. The high purities allow the successful engraftment of immunodeficient mice without further purification and may speed up the development of allogeneic V?2V?2 T cell therapies derived from HLA-matched normal donors for patients with poor autologous V?2V?2 T cell responses.

SUBMITTER: Tanaka Y

PROVIDER: S-EPMC5834800 | biostudies-other | 2018 Mar

REPOSITORIES: biostudies-other

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Expansion of human γδ T cells for adoptive immunotherapy using a bisphosphonate prodrug.

Tanaka Yoshimasa Y Murata-Hirai Kaoru K Iwasaki Masashi M Matsumoto Kenji K Hayashi Kosuke K Kumagai Asuka A Nada Mohanad H MH Wang Hong H Kobayashi Hirohito H Kamitakahara Hiroshi H Okamura Haruki H Sugie Tomoharu T Minato Nagahiro N Toi Masakazu M Morita Craig T CT

Cancer science 20180204 3

Cancer immunotherapy with human γδ T cells expressing Vγ2Vδ2 T cell receptor (also termed Vγ9Vδ2) has shown promise because of their ability to recognize and kill most types of tumors in a major histocombatibility complex (MHC) -unrestricted fashion that is independent of the number of tumor mutations. In clinical trials, adoptive transfer of Vγ2Vδ2 T cells has been shown to be safe and does not require preconditioning. In this report, we describe a method for preparing highly enriched human Vγ2 ...[more]

PMID: 29288540

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Project description:Increasing attention has been paid to human γδ T cells expressing Vγ2Vδ2 T cell receptor (also termed Vγ9Vδ2) in the field of cancer immunotherapy. We have previously demonstrated that a novel bisphosphonate prodrug, tetrakis-pivaloyloxymethyl 2-(thiazole-2-ylamino)ethylidene-1,1-bisphosphonate (PTA), efficiently expands peripheral blood Vγ2Vδ2 T cells to purities up to 95-99% in 10-11 days. In the present study, we first examined the effect of PTA on farnesyl diphosphate synthase (FDPS) using liquid chromatography mass spectrometry (LC-MS) to analyze the mechanism underlying the PTA-mediated expansion of Vγ2Vδ2 T cells. We find that the prodrug induced the accumulation of both isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), direct upstream metabolites of FDPS. This indicates that not only IPP but also DMAPP plays an important role in PTA-mediated stimulation of Vγ2Vδ2 T cells. We next analyzed TCR-independent cytotoxicity of Vγ2Vδ2 T cells. When human lung cancer cell lines were challenged by Vγ2Vδ2 T cells, no detectable cytotoxicity was observed in 40 min. The lung cancer cell lines were, however, significantly killed by Vγ2Vδ2 T cells after 4-16 h in an effector-to-target ratio-dependent manner, demonstrating that Vγ2Vδ2 T cell-based cell therapy required a large number of cells and longer time when tumor cells were not sensitized. By contrast, pulsing tumor cell lines with 10-30 nM of PTA induced significant lysis of tumor cells by Vγ2Vδ2 T cells even in 40 min. Similar levels of cytotoxicity were elicited by ZOL at concentrations of 100-300 μM, which were much higher than blood levels of ZOL after infusion (1-2 μM), suggesting that standard 4 mg infusion of ZOL was not enough to sensitize lung cancer cells in clinical settings. In addition, Vγ2Vδ2 T cells secreted interferon-γ (IFN-γ) when challenged by lung cancer cell lines pulsed with PTA in a dose-dependent manner. Taken together, PTA could be utilized for both expansion of Vγ2Vδ2 T cells ex vivo and sensitization of tumor cells in vivo in Vγ2Vδ2 T cell-based cancer immunotherapy. For use in patients, further studies on drug delivery are essential because of the hydrophobic nature of the prodrug.

Dataset Information

Expansion of human ?? T cells for adoptive immunotherapy using a bisphosphonate prodrug.

Publications

Expansion of human γδ T cells for adoptive immunotherapy using a bisphosphonate prodrug.

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