In vivo intracellular oxygen dynamics in murine brain glioma and immunotherapeutic response of cytotoxic T cells observed by fluorine-19 magnetic resonance imaging.

Jia Zhong,Masashi Sakaki,Hideho Okada,Eric T Ahrens

doi:10.1371/journal.pone.0059479

Abstract

Noninvasive biomarkers of anti-tumoral efficacy are of great importance to the development of therapeutic agents. Tumor oxygenation has been shown to be an important indicator of therapeutic response. We report the use of intracellular labeling of tumor cells with perfluorocarbon (PFC) molecules, combined with quantitative 19F spin-lattice relaxation rate (R1) measurements, to assay tumor cell oxygen dynamics in situ. In a murine central nervous system (CNS) GL261 glioma model, we visualized the impact of Pmel-1 cytotoxic T cell immunotherapy, delivered intravenously, on intracellular tumor oxygen levels. GL261 glioma cells were labeled ex vivo with PFC and inoculated into the mouse striatum. The R1 of 19F labeled cells was measured using localized single-voxel magnetic resonance spectroscopy, and the absolute intracellular partial pressure of oxygen (pO2) was ascertained. Three days after tumor implantation, mice were treated with 2×107 cytotoxic T cells intravenously. At day five, a transient spike in pO2 was observed indicating an influx of T cells into the CNS and putative tumor cell apoptosis. Immunohistochemistry and quantitative flow cytometry analysis confirmed that the pO2 was causally related to the T cells infiltration. Surprisingly, the pO2 spike was detected even though few (∼4×104) T cells actually ingress into the CNS and with minimal tumor shrinkage. These results indicate the high sensitivity of this approach and its utility as a non-invasive surrogate biomarker of anti-cancer immunotherapeutic response in preclinical models.

Highlights

Malignant gliomas are the most common type of primary brain tumor and a significant public health problem, with more than 14,000 new cases diagnosed each year in the US [1]
Immunohistochemistry (IHC) and flow cytometry analysis were used to further confirm our in vivo findings. These data show that an increase in pO2 can reliably be observed, even though relatively few T cells ingress into the central nervous system (CNS) tumor
This study demonstrates, for the first time, the dynamics of the tumor intracellular pO2 in vivo following treatment with tumor infiltrating lymphocytes (TILs)

Summary

Introduction

Malignant gliomas are the most common type of primary brain tumor and a significant public health problem, with more than 14,000 new cases diagnosed each year in the US [1]. Glioblastoma multiforme (GBM) is by far the most common and most malignant of the glial tumors. Recent clinical trial results show that concomitant temozolomide and radiotherapy improved the 2-year survival to 27.2% compared to only a 10.2% survival rate for patients receiving radiation therapy alone [2]. This trial demonstrates a significant advancement in the treatment of GBM, novel strategies are required to better treat patients with this aggressive cancer

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