Characterization of a novel murine model for spontaneous hemorrhagic stroke using in vivo PET and MR multiparametric imaging

ABSTRACT The clinical use of Magnetic Resonance Imaging (MRI) and Positron Emission Tomography (PET) has proven to be a strong diagnostic tool in the field of neurology. The reliability of these methods to confirm clinical diagnoses has guided preclinical research to utilize these techniques for the characterization of animal disease models. Previously, we demonstrated that an endothelial cell‐specific ablation of the murine Serum Response Factor (Srf iECKO) results in blood brain barrier (BBB) breakdown and hemorrhagic stroke. Taking advantage of this mouse model we here perform a comprehensive longitudinal, multiparametric and in vivo imaging approach to reveal pathophysiological processes occurring before and during the appearance of cerebral microbleeds using combined PET and MRI. We complement our imaging results with data regarding animal behavior and immunohistochemistry. Our results demonstrate diffusion abnormalities in the cortical brain tissue prior to the onset of cerebral microbleeds. Diffusion reductions were accompanied by significant increments of [18F]FAZA uptake before the onset of the lesions in T2WI. The Open Field behavioral tests revealed reduced activity of Srf iECKO animals, whereas histology confirmed the presence of hemorrhages in cortical regions of the mouse brain and iron deposition at lesion sites with increased hypoxia inducible factor 1&agr;, CD31 and glial fibrillary acidic protein expression. For the first time, we performed a thorough evaluation of the prodromal period before the occurrence of spontaneous cerebral microbleeds. Using in vivo PET and MRI, we show the pathological tissue changes that occur previous to gross blood brain barrier (BBB) disruption and breakage. In addition, our results show that apparent diffusion coefficient (ADC) reduction may be an early biomarker of BBB disruption proposing an alternate clinical interpretation. Furthermore, our findings remark the usefulness of this novel Srf iECKO mouse model to study underlying mechanisms of hemorrhagic stroke. HIGHLIGHTSPET‐MRI characterization of an endogenous spontaneous stroke modelBBB breakage courses with reductions in water diffusionEvolution of microangiopathies not associated to amyloid plaque deposition.

[1]  T. Videen,et al.  Transient focal increase in perihematomal glucose metabolism after acute human intracerebral hemorrhage. , 2009, Stroke.

[2]  M. Hoehn,et al.  Dynamic changes of magnetic resonance imaging abnormalities in relation to inflammation and glial responses after photothrombotic cerebral infarction in the rat brain , 2001, Acta Neuropathologica.

[3]  Habib Zaidi,et al.  Clinical Applications of Hybrid PET/MRI in Neuroimaging , 2013, Clinical nuclear medicine.

[4]  S. Homer-Vanniasinkam,et al.  Relationship Between Leukocyte Kinetics and Behavioral Tests Changes in the Inflammatory Process of Hemorrhagic Stroke Recovery , 2010, The International journal of neuroscience.

[5]  Ciprian Catana,et al.  PET/MRI for Neurologic Applications , 2012, The Journal of Nuclear Medicine.

[6]  M. Alscher,et al.  Störungen des Säure-Basen-Haushaltes und der Anionenlücke , 2016, DMW - Deutsche Medizinische Wochenschrift.

[7]  J. Baron,et al.  A comparison of four PET tracers for brain hypoxia mapping in a rodent model of stroke. , 2013, Nuclear medicine and biology.

[8]  A. Schmid,et al.  Quantification accuracy and partial volume effect in dependence of the attenuation correction of a state-of-the-art small animal PET scanner , 2012, Physics in medicine and biology.

[9]  C. Beaulieu,et al.  Use of diffusion weighted MRI to predict the occurrence and severity of hemorrhagic transformation in a rabbit model of embolic stroke , 2002, Brain Research.

[10]  S. Williams,et al.  Diffusion‐Weighted Imaging Studies of Cerebral Ischemia in Gerbils: Potential Relevance to Energy Failure , 1992, Stroke.

[11]  J. Pipe,et al.  Evidence for cytotoxic edema in the pathogenesis of cerebral venous infarction. , 2001, AJNR. American journal of neuroradiology.

[12]  D. Donner,et al.  Vascular Endothelial Cell Growth Factor Promotes Tyrosine Phosphorylation of Mediators of Signal Transduction That Contain SH2 Domains , 1995, The Journal of Biological Chemistry.

[13]  Stefan Wiehr,et al.  Longitudinal PET-MRI reveals β-amyloid deposition and rCBF dynamics and connects vascular amyloidosis to quantitative loss of perfusion , 2014, Nature Medicine.

[14]  S. Di Giovanni,et al.  Serum Response Factor (SRF)-cofilin-actin signaling axis modulates mitochondrial dynamics , 2012, Proceedings of the National Academy of Sciences.

[15]  Levick Capillary filtration‐absorption balance reconsidered in light of dynamic extravascular factors , 1991, Experimental physiology.

[16]  G. Manley,et al.  Aquaporin‐4 facilitates reabsorption of excess fluid in vasogenic brain edema , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[17]  A S Verkman,et al.  Three distinct roles of aquaporin-4 in brain function revealed by knockout mice. , 2006, Biochimica et biophysica acta.

[18]  E. J. Green,et al.  Sensorimotor and cognitive consequences of middle cerebral artery occlusion in rats , 1992, Brain Research.

[19]  Alfred Nordheim,et al.  Linking actin dynamics and gene transcription to drive cellular motile functions , 2010, Nature Reviews Molecular Cell Biology.

[20]  Jeannette M. Perez-Rossello,et al.  In vivo assessment of optimal b-value range for perfusion-insensitive apparent diffusion coefficient imaging. , 2012, Medical physics.

[21]  Gereon R Fink,et al.  Neuroinflammation Extends Brain Tissue at Risk to Vital Peri-Infarct Tissue: A Double Tracer [11C]PK11195- and [18F]FDG-PET Study , 2009, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[22]  A. Nordheim,et al.  Serum response factor is crucial for actin cytoskeletal organization and focal adhesion assembly in embryonic stem cells , 2002, The Journal of cell biology.

[23]  C. Calaminus,et al.  Endothelial depletion of murine SRF/MRTF provokes intracerebral hemorrhagic stroke , 2015, Proceedings of the National Academy of Sciences.

[24]  H. Pasolli,et al.  Developmental roles for Srf, cortical cytoskeleton and cell shape in epidermal spindle orientation , 2011, Nature Cell Biology.

[25]  Erika Gustafsson,et al.  Serum response factor is required for sprouting angiogenesis and vascular integrity. , 2008, Developmental cell.

[26]  U. Klose,et al.  FAIR true‐FISP perfusion imaging of the kidneys , 2004, Magnetic resonance in medicine.

[27]  Bruno Brochet,et al.  Aquaporin 4 correlates with apparent diffusion coefficient and hydrocephalus severity in the rat brain: A combined MRI–histological study , 2009, NeuroImage.

[28]  C. Solbach,et al.  Preparation of the hypoxia imaging PET tracer [18F]FAZA: reaction parameters and automation. , 2005, Applied radiation and isotopes : including data, instrumentation and methods for use in agriculture, industry and medicine.

[29]  A. Schmid,et al.  Significant impact of different oxygen breathing conditions on noninvasive in vivo tumor-hypoxia imaging using [18F]-fluoro-azomycinarabino-furanoside ([18F]FAZA) , 2011, Radiation oncology.

[30]  T. Beyer,et al.  Variations in PET/MRI Operations: Results from an International Survey Among 39 Active Sites , 2016, The Journal of Nuclear Medicine.

[31]  A. Nordheim,et al.  Serum response factor is essential for mesoderm formation during mouse embryogenesis , 1998, The EMBO journal.

[32]  P. Yates,et al.  Intracerebral microaneurysms and small cerebrovascular lesions. , 1967, Brain : a journal of neurology.

[33]  S. Mayer,et al.  Multimodality Monitoring for Cerebral Perfusion Pressure Optimization in Comatose Patients With Intracerebral Hemorrhage , 2011, Stroke.

[34]  J C Froment,et al.  Old Microbleeds Are a Potential Risk Factor for Cerebral Bleeding After Ischemic Stroke: A Gradient-Echo T2*-Weighted Brain MRI Study , 2002, Stroke.

[35]  Haibin Shi,et al.  Hypertension increases the risk of cerebral microbleed in the territory of posterior cerebral artery: a study of the association of microbleeds categorized on a basis of vascular territories and cardiovascular risk factors. , 2014, Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association.

[36]  J. Kucharczyk,et al.  Diffusion-Weighted Magnetic Resonance Imaging of Acute Focal Cerebral Ischemia: Comparison of Signal Intensity with Changes in Brain Water and Na+,K+ -ATPase Activity , 1994, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[37]  I. Loubinoux,et al.  Spreading of vasogenic edema and cytotoxic edema assessed by quantitative diffusion and T2 magnetic resonance imaging. , 1997, Stroke.

[38]  S. Sarubbo,et al.  Temporal changes in perihematomal apparent diffusion coefficient values during the transition from acute to subacute phases in patients with spontaneous intracerebral hemorrhage , 2013, Neuroradiology.

[39]  G. Dobson,et al.  Intracellular, interstitial and plasma spaces in the rat myocardium in vivo. , 1997, Journal of molecular and cellular cardiology.

[40]  S. Jakobsen,et al.  PET hypoxia imaging with FAZA: reproducibility at baseline and during fractionated radiotherapy in tumour-bearing mice , 2012, European Journal of Nuclear Medicine and Molecular Imaging.

[41]  I. Módy,et al.  Novel test of motor and other dysfunctions in mouse neurological disease models , 2014, Journal of Neuroscience Methods.

[42]  George Paxinos,et al.  The Mouse Brain in Stereotaxic Coordinates , 2001 .

[43]  E. Rickels Middle Cerebral Artery Occlusion in Rats , 1993 .

[44]  A. Konstas,et al.  Reperfusion injury following cerebral ischemia: pathophysiology, MR imaging, and potential therapies , 2006, Neuroradiology.

[45]  Mickael Tanter,et al.  Imaging of Perfusion, Angiogenesis, and Tissue Elasticity after Stroke , 2012, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[46]  J. Pauls,et al.  Neuroradiological findings in vascular dementia , 2006, Neuroradiology.

[47]  P. Grenier,et al.  MR imaging of intravoxel incoherent motions: application to diffusion and perfusion in neurologic disorders. , 1986, Radiology.

[48]  L. Hedlund,et al.  Mechanism of Detection of Acute Cerebral Ischemia in Rats by Diffusion‐Weighted Magnetic Resonance Microscopy , 1992, Stroke.

[49]  Jean-Claude Baron,et al.  Section Editors: Wolf- Applications of Nitroimidazole in Vivo Hypoxia Imaging in Ischemic Stroke , 2022 .

[50]  K. Hamacher,et al.  Efficient stereospecific synthesis of no-carrier-added 2-[18F]-fluoro-2-deoxy-D-glucose using aminopolyether supported nucleophilic substitution. , 1986, Journal of nuclear medicine : official publication, Society of Nuclear Medicine.

[51]  R. E. Adams,et al.  The effects of mannitol on cerebral edema after large hemispheric cerebral infarct , 1999, Neurology.

[52]  A. Nordheim,et al.  Generation of mice carrying conditional knockout alleles for the transcription factor SRF , 2002, Genesis.

[53]  T. Duong,et al.  T2*-weighted fMRI time-to-peak of oxygen challenge in ischemic stroke , 2016, Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism.

[54]  R. Kim,et al.  Cerebral Microbleeds in the Elderly: A Pathological Analysis , 2010, Stroke.

[55]  E. Mark Haacke,et al.  Correlation of hypointensities in susceptibility-weighted images to tissue histology in dementia patients with cerebral amyloid angiopathy: a postmortem MRI study , 2010, Acta Neuropathologica.

[56]  S. Beck,et al.  Endothelial SRF/MRTF ablation causes vascular disease phenotypes in murine retinae. , 2013, The Journal of clinical investigation.

[57]  J. Neil Measurement of water motion (apparent diffusion) in biologial systems , 1997 .

[58]  G. Griebel,et al.  Measuring normal and pathological anxiety-like behaviour in mice: a review , 2001, Behavioural Brain Research.

[59]  W. Hacke,et al.  Hemorrhagic Transformation of Ischemic Brain Tissue: Asymptomatic or Symptomatic? , 2001, Stroke.

[60]  S. Taheri,et al.  Spatiotemporal Correlations between Blood-Brain Barrier Permeability and Apparent Diffusion Coefficient in a Rat Model of Ischemic Stroke , 2009, PloS one.