Accumulation of hypoxia imaging probe “18F-FMISO” in macrophages depends on macrophage polarization in addition to hypoxic state
Abstract
Objective
Macrophages play an essential role in immune response, and are closely related to the progression of diseases such as cancer and atherosclerosis. Macrophages polarize to M1 or M2 type, which is related to the environmental hypoxic state. Previously, we found that 18F-FMISO uptake varied according to expression levels of biomolecules such as glutathione S-transferase P1 (GST-P1), which catalyzes the conjugation of glutathione to 18F-FMISO metabolites, and multidrug resistance-associated protein 1 (MRP1), which exports glutathione-18F-FMISO metabolite conjugates out of cells. However, the relationship between macrophage polarization and 18F-FMISO accumulation remains unclear.
Methods
Mouse peritoneal macrophages were polarized to either the M1 or M2 type, and were treated with 18F-FMISO. Then, their radioactivity after a 4 h incubation period under normoxic (21% O2) or hypoxic (1% O2) condition was measured. GST-P1 and MRP1 expression levels were measured by qRT-PCR.
Results
M2 macrophages exhibited a significantly higher uptake of 18F-FMISO than non-polarized (M0) macrophages, whereas M1 macrophages had a significantly lower uptake than M0 macrophages (M0: 1.05 ± 0.22, M1: 0.34 ± 0.02, M2: 4.17 ± 0.36 %dose/mg protein). The GST-P1 expression level in M1 macrophages was higher than that in M2 and M0 macrophages [GST-P1/β-actin normalized by M0: 9.0 ± 3.7 (M1), 1.2 ± 0.2 (M2)]. The MRP1 expression level in M1 macrophages was significantly higher than that in M2 and M0 macrophages [MRP1/β-actin normalized by M0 macrophages: 5.1 ± 2.1 (M1), 2.8 ± 1.0 (M2)].
Conclusions
18F-FMISO accumulation in macrophages may depend on the polarization state in addition to hypoxic condition.
Keywords
18F-FMISO Macrophage Hypoxia Polarization M1 M2Notes
Acknowledgements
The authors thank the staff of the Hokkaido University Hospital Cyclotron Facility for the synthesis of 18F-FMISO. This study was supported in part by JSPS KAKENHI (Grant number: 16K19799).
Supplementary material
References
- 1.Gordon S, Taylor PR. Monocyte and macrophage heterogeneity. Nat Rev Immunol. 2005;5:953–64.CrossRefGoogle Scholar
- 2.Sica A, Mantovani A. Macrophage plasticity and polarization: in vivo veritas. J Clin Investig. 2012;122:787–95.CrossRefGoogle Scholar
- 3.Shapouri-Moghaddam A, Mohammadian S, Vazini H, Taghadosi M, Esmaeili SA, Mardani F, et al. Macrophage plasticity, polarization, and function in health and disease. J Cell Physiol. 2018;233:6425–40.CrossRefGoogle Scholar
- 4.Chinetti-Gbaguidi G, Baron M, Bouhlel MA, Vanhoutte J, Copin C, Sebti Y, et al. Human atherosclerotic plaque alternative macrophages display low cholesterol handling but high phagocytosis because of distinct activities of the PPARgamma and LXRalpha pathways. Circ Res. 2011;108:985–95.CrossRefGoogle Scholar
- 5.Zheng X, Turkowski K, Mora J, Brune B, Seeger W, Weigert A, et al. Redirecting tumor-associated macrophages to become tumoricidal effectors as a novel strategy for cancer therapy. Oncotarget. 2017;8:48436–52.Google Scholar
- 6.Henze AT, Mazzone M. The impact of hypoxia on tumor-associated macrophages. J Clin Investig. 2016;126:3672–9.CrossRefGoogle Scholar
- 7.Lopci E, Grassi I, Chiti A, Nanni C, Cicoria G, Toschi L, et al. PET radiopharmaceuticals for imaging of tumor hypoxia: a review of the evidence. Am J Nucl Med Mol Imaging. 2014;4:365–84.Google Scholar
- 8.Masaki Y, Shimizu Y, Yoshioka T, Nishijima KI, Zhao S, Higashino K, et al. FMISO accumulation in tumor is dependent on glutathione conjugation capacity in addition to hypoxic state. Ann Nucl Med. 2017;31:596–604.CrossRefGoogle Scholar
- 9.Masaki Y, Shimizu Y, Yoshioka T, Tanaka Y, Nishijima K, Zhao S, et al. The accumulation mechanism of the hypoxia imaging probe “FMISO” by imaging mass spectrometry: possible involvement of low-molecular metabolites. Sci Rep. 2015;5:16802.CrossRefGoogle Scholar
- 10.Ogawa M, Nakamura S, Saito Y, Kosugi M, Magata Y. What can be seen by 18F-FDG PET in atherosclerosis imaging? The effect of foam cell formation on 18F-FDG uptake to macrophages in vitro. J Nucl Med. 2012;53:55–8.CrossRefGoogle Scholar
- 11.Shimizu Y, Hanzawa H, Zhao Y, Fukura S, Nishijima KI, Sakamoto T, et al. Immunoglobulin G (IgG)-based imaging probe accumulates in m1 macrophage-infiltrated atherosclerotic plaques independent of igg target molecule expression. Mol Imaging Biol. 2017;19:531–9.CrossRefGoogle Scholar
- 12.Shimizu Y, Zhao S, Yasui H, Nishijima KI, Matsumoto H, Shiga T, et al. A novel PET probe “[18F]DiFA” accumulates in hypoxic region via glutathione conjugation following reductive metabolism. Mol Imaging Biol. 2018. https://doi.org/10.1007/s11307-018-1214-y.Google Scholar
- 13.Sekine H, Yamamoto M, Motohashi H. Tumors sweeten macrophages with acids. Nat Immunol. 2018;19:1281–3.CrossRefGoogle Scholar
- 14.Kovacevic Z, Sahni S, Lok H, Davies MJ, Wink DA, Richardson DR. Regulation and control of nitric oxide (NO) in macrophages: Protecting the “professional killer cell” from its own cytotoxic arsenal via MRP1 and GSTP1. Biochim Biophys Acta Gen Subj. 2017;1861:995–9.CrossRefGoogle Scholar
- 15.Colegio OR, Chu NQ, Szabo AL, Chu T, Rhebergen AM, Jairam V, et al. Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature. 2014;513:559–63.CrossRefGoogle Scholar
- 16.Bohn T, Rapp S, Luther N, Klein M, Bruehl TJ, Kojima N, et al. Tumor immunoevasion via acidosis-dependent induction of regulatory tumor-associated macrophages. Nat Immunol. 2018;19:1319–29.CrossRefGoogle Scholar