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1H MRS detects polyunsaturated fatty acid accumulation during gene therapy of glioma: Implications for the in vivo detection of apoptosis

Nature Medicine volume 5, pages 1323–1327 (1999)Cite this article

Cancer cells typically die via apoptosis both upon conventional chemotherapy, radiation1, and as result of experimental approaches such as antiangiogenic2 and ganciclovir (GCV) treatment of herpes simplex thymidine kinase (HSV-tk)-transfected cells3,4. Consequently, it has been suggested that facilitation of apoptosis by selective chemotherapeutic agents or gene therapy could be a target for human cancer treatment in the future1. This has generated wide interest in the multitude of biochemical and molecular events involved in programmed cell death. Many of these processes have essentially been described in in vitro systems, and their applicability to complex and heterogeneous in vivo systems remains to be established. This has largely been caused by the lack of specific methods to address this issue in vivo.

Magnetic resonance imaging (MRI) has recently been exploited to monitor tumor treatment non-invasively in chemotherapy and HSV-tk-mediated therapy schemes in vivo. Prior to volume loss, treatment response is associated with an increase in tissue water diffusion5 and T2 relaxation time5, suggesting increased water content and bulk diffusivity. Recently, reduced diffusion of choline (Cho)-containing compounds in gliomas undergoing apoptosis was detected6. This observation implies increased viscosity and restriction within cells, and appears to mirror cell shrinkage. Inasmuch as physiological properties of tissue primarily affect MRI contrast, relaxation and diffusion parameters may actually reflect morphological events, such as cell shrinkage, membrane blebbing, and the shedding of apoptotic bodies7, occurring in the late stages of the apoptotic process.

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Figure 1: Tissue response in BT4C glioma sections (×60) stained with TUNEL to identify apoptotic cells, counterstained with methyl green.
Figure 2: Transmission electron micrographs from BT4C glioma tissue (×5,000) fixed with OsO4 to identify intracellular lipids.
Figure 3: A typical time course of 1H MRS spectral changes from one GCV-treated animal in vivo.
Figure 4: Time courses of in vivo MRS resonances during GCV treatment.

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Acknowledgements

We thank Kati Holopainen and Mervi Nieminen for expert technical assistance, Alpo Pelttari for sharing his knowledge in electron microscopy, and Helvi Vidgren and Arja Erkkilä for co-analyzing tissue lipids. This study was funded by the Finnish Cancer Foundation and Sigrid Juselius Foundation (R.A.K.), Academy of Finland (J.M.H., S.Y.H., R.A.K.), Emil Aaltonen Foundation, Finnish Medical Foundation, Northern Savo Cancer Fund and The North Savo Fund of Finnish Cultural Foundation (J.M.H.).

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  1. Harish Poptani

    Present address: H.P. Present address: Department of Radiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA,

Authors and Affiliations

  1. NMR Research Group, A.I.Virtanen Institute for Molecular Sciences, University of Kuopio, Kuopio, FIN-70211, Finland

    Juhana M. Hakumäki, Harish Poptani & Risto A. Kauppinen

  2. Molecular Medicine Group, A.I.Virtanen Institute for Molecular Sciences, University of Kuopio, Kuopio, FIN-70211, Finland

    Anu-Maaria Sandmair & Seppo Ylä-Herttuala

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  1. Juhana M. Hakumäki
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Correspondence to Juhana M. Hakumäki or Risto A. Kauppinen.

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Hakumäki, J., Poptani, H., Sandmair, AM. et al. 1H MRS detects polyunsaturated fatty acid accumulation during gene therapy of glioma: Implications for the in vivo detection of apoptosis. Nat Med 5, 1323–1327 (1999). https://doi.org/10.1038/15279

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