title: Glial activation in the early stages of brain metastasis: TSPO as a diagnostic biomarker.
abstract: Metastatic spread of cancer cells to the brain is associated with high mortality, primarily because current diagnostic tools identify only well-advanced metastases. Brain metastases have been shown to induce a robust glial response, including both astrocyte and microglial activation. On the basis of these findings, we hypothesized that this stromal response may provide a sensitive biomarker of tumor burden, in particular through the use of SPECT/PET imaging agents targeting the translocator protein (TSPO) that is upregulated on activated glia. Our goals, therefore, were first to determine the spatial and temporal profile of glial activation during early metastasis growth in vivo and second to assess the potential of the radiolabeled TSPO ligand (123)I-DPA-713 for early detection of brain metastases.
  
  
  METHODS
  Metastatic mouse mammary carcinoma 4T1-green fluorescent protein cells were injected either intracerebrally or intracardially into female BALB/c mice to induce brain metastases. Astrocyte and microglial activation was assessed immunohistochemically over a 28-d period, together with immunofluorescence detection of TSPO upregulation. Subsequently, SPECT imaging and autoradiography were used to determine in vivo binding of (123)I-DPA-713 at metastatic sites.
  
  
  RESULTS
  Dynamic astrocyte and microglial activation was evident throughout the early stages of tumor growth, with the extent of astrocyte activation correlating significantly with tumor size (P < 0.0001). Microglial activation appeared to increase more rapidly than astrocyte activation at the earlier time points, but by later time points the extent of activation was comparable between the glial cell types. Upregulation of TSPO expression was found on both glial populations. Both autoradiographic and in vivo SPECT data showed strong positive binding of (123)I-DPA-713 in the intracerebrally induced model of brain metastasis, which was significantly greater than that observed in controls (P < 0.05). (123)I-DPA-713 binding was also evident autoradiographically in the intracardially induced model of brain metastasis but with lower sensitivity because of smaller tumor size (∼ 100-μm diameter vs. ∼ 600-μm diameter in the intracerebral model).
  
  
  CONCLUSIONS
  These data suggest that the glial response to brain metastasis may provide a sensitive biomarker of tumor burden, with a tumor detection threshold lying between 100 and 600 μm in diameter. This approach could enable substantially earlier detection of brain metastases than the current clinical approach of gadolinium-enhanced MR imaging.
authors.affiliation: CR-United Kingdom/MRC Gray Institute for Radiation Oncology and Biology, Department of Oncology, University of Oxford, Churchill Hospital, Oxford, United Kingdom.
authors.forename: Emma R
authors.initials: ER
authors.lastname: O'Brien
authors.forename: Veerle
authors.initials: V
authors.lastname: Kersemans
authors.forename: Matthew
authors.initials: M
authors.lastname: Tredwell
authors.forename: Begona
authors.initials: B
authors.lastname: Checa
authors.forename: Sébastien
authors.initials: S
authors.lastname: Serres
authors.forename: Manuel Sarmiento
authors.initials: MS
authors.lastname: Soto
authors.forename: Veronique
authors.initials: V
authors.lastname: Gouverneur
authors.forename: David
authors.initials: D
authors.lastname: Leppert
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Anthony
authors.forename: Nicola R
authors.initials: NR
authors.lastname: Sibson
mesh_terms: D000081:Acetamides; D000818:Animals; D001253:Astrocytes / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000378:metabolism; D001932:Brain Neoplasms / Q000000981:diagnostic imaging* / Q000378:metabolism*; D005260:Female; D005682:Gadolinium / Q000737:chemistry; D015972:Gene Expression Regulation, Neoplastic; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D008024:Ligands; D008279:Magnetic Resonance Imaging; D051379:Mice; D008807:Mice, Inbred BALB C; D017628:Microglia / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009362:Neoplasm Metastasis / Q000175:diagnosis*; D009368:Neoplasm Transplantation; D009457:Neuroglia / Q000000981:diagnostic imaging / Q000378:metabolism*; D011485:Protein Binding; D011720:Pyrazoles; D011743:Pyrimidines; D018079:Receptors, GABA / Q000378:metabolism*; D013997:Time Factors; D015899:Tomography, Emission-Computed, Single-Photon
chemical_list: D000081:Acetamides; D014408:Biomarkers, Tumor; C463819:Bzrp protein, mouse; D008024:Ligands; C507091:N,N-diethyl-2-(2-(4-methoxyphenyl)-5,7-dimethyl-pyrazolo(1,5-a)pyrimidin-3-yl)-acetamide; D011720:Pyrazoles; D011743:Pyrimidines; D018079:Receptors, GABA; D049452:Green Fluorescent Proteins; D005682:Gadolinium
keywords: SPECT imaging; TSPO; astrocytes; metastasis; microglia; mouse